Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies

PubMed Central

Saylor, Karen L.; Anver, Miriam R.; Salomon, David S.; Golubeva, Yelena G.

2016-01-01

Laser capture microdissection (LCM) of tissue is an established tool in medical research for collection of distinguished cell populations under direct microscopic visualization for molecular analysis. LCM samples have been successfully analyzed in a number of genomic and proteomic downstream molecular applications. However, LCM sample collection and preparation procedure has to be adapted to each downstream analysis platform. In this present manuscript we describe in detail the adaptation of LCM methodology for the collection and preparation of fresh frozen samples for NanoString analysis based on a study of a model of mouse mammary gland carcinoma and its lung metastasis. Our adaptation of LCM sample preparation and workflow to the requirements of the NanoString platform allowed acquiring samples with high RNA quality. The NanoString analysis of such samples provided sensitive detection of genes of interest and their associated molecular pathways. NanoString is a reliable gene expression analysis platform that can be effectively coupled with LCM. PMID:27077656

Microfluidic Sample Preparation for Diagnostic Cytopathology

PubMed Central

Mach, Albert J.; Adeyiga, Oladunni B.; Di Carlo, Dino

2014-01-01

The cellular components of body fluids are routinely analyzed to identify disease and treatment approaches. While significant focus has been placed on developing cell analysis technologies, tools to automate the preparation of cellular specimens have been more limited, especially for body fluids beyond blood. Preparation steps include separating, concentrating, and exposing cells to reagents. Sample preparation continues to be routinely performed off-chip by technicians, preventing cell-based point-of-care diagnostics, increasing the cost of tests, and reducing the consistency of the final analysis following multiple manually-performed steps. Here, we review the assortment of biofluids for which suspended cells are analyzed, along with their characteristics and diagnostic value. We present an overview of the conventional sample preparation processes for cytological diagnosis. We finally discuss the challenges and opportunities in developing microfluidic devices for the purpose of automating or miniaturizing these processes, with particular emphases on preparing large or small volume samples, working with samples of high cellularity, automating multi-step processes, and obtaining high purity subpopulations of cells. We hope to convey the importance of and help identify new research directions addressing the vast biological and clinical applications in preparing and analyzing the array of available biological fluids. Successfully addressing the challenges described in this review can lead to inexpensive systems to improve diagnostic accuracy while simultaneously reducing overall systemic healthcare costs. PMID:23380972

GUM Analysis for TIMS Isotopic Ratios in BEP0 Graphite Qualification Samples, Round 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gerlach, David C.; Heasler, Patrick G.; Reid, Bruce D.

In May 2007, one set of three samples from NBL were addressed to Steve Petersen for TIMS analysis, and included BEP0 samples numbered 27008, 30986, and 50846. All cores were trimmed by tooling, and lightly cleaned by CO2 pellet blasting. Small discs were cut from the second set of samples for SIMS analysis, with the remainder of each used for TIMS preparation.

Development of analytical cell support for vitrification at the West Valley Demonstration Project. Topical report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barber, F.H.; Borek, T.T.; Christopher, J.Z.

1997-12-01

Analytical and Process Chemistry (A&PC) support is essential to the high-level waste vitrification campaign at the West Valley Demonstration Project (WVDP). A&PC characterizes the waste, providing information necessary to formulate the recipe for the target radioactive glass product. High-level waste (HLW) samples are prepared and analyzed in the analytical cells (ACs) and Sample Storage Cell (SSC) on the third floor of the main plant. The high levels of radioactivity in the samples require handling them in the shielded cells with remote manipulators. The analytical hot cells and third floor laboratories were refurbished to ensure optimal uninterrupted operation during the vitrificationmore » campaign. New and modified instrumentation, tools, sample preparation and analysis techniques, and equipment and training were required for A&PC to support vitrification. Analytical Cell Mockup Units (ACMUs) were designed to facilitate method development, scientist and technician training, and planning for analytical process flow. The ACMUs were fabricated and installed to simulate the analytical cell environment and dimensions. New techniques, equipment, and tools could be evaluated m in the ACMUs without the consequences of generating or handling radioactive waste. Tools were fabricated, handling and disposal of wastes was addressed, and spatial arrangements for equipment were refined. As a result of the work at the ACMUs the remote preparation and analysis methods and the equipment and tools were ready for installation into the ACs and SSC m in July 1995. Before use m in the hot cells, all remote methods had been validated and four to eight technicians were trained on each. Fine tuning of the procedures has been ongoing at the ACs based on input from A&PC technicians. Working at the ACs presents greater challenges than had development at the ACMUs. The ACMU work and further refinements m in the ACs have resulted m in a reduction m in analysis turnaround time (TAT).« less

Lab-on-a-chip modules for detection of highly pathogenic bacteria: from sample preparation to detection

NASA Astrophysics Data System (ADS)

Julich, S.; Kopinč, R.; Hlawatsch, N.; Moche, C.; Lapanje, A.; Gärtner, C.; Tomaso, H.

2014-05-01

Lab-on-a-chip systems are innovative tools for the detection and identification of microbial pathogens in human and veterinary medicine. The major advantages are small sample volume and a compact design. Several fluidic modules have been developed to transform analytical procedures into miniaturized scale including sampling, sample preparation, target enrichment, and detection procedures. We present evaluation data for single modules that will be integrated in a chip system for the detection of pathogens. A microfluidic chip for purification of nucleic acids was established for cell lysis using magnetic beads. This assay was evaluated with spiked environmental aerosol and swab samples. Bacillus thuringiensis was used as simulant for Bacillus anthracis, which is closely related but non-pathogenic for humans. Stationary PCR and a flow-through PCR chip module were investigated for specific detection of six highly pathogenic bacteria. The conventional PCR assays could be transferred into miniaturized scale using the same temperature/time profile. We could demonstrate that the microfluidic chip modules are suitable for the respective purposes and are promising tools for the detection of bacterial pathogens. Future developments will focus on the integration of these separate modules to an entire lab-on-a-chip system.

Sample preparation prior to the LC-MS-based metabolomics/metabonomics of blood-derived samples.

PubMed

Gika, Helen; Theodoridis, Georgios

2011-07-01

Blood represents a very important biological fluid and has been the target of continuous and extensive research for diagnostic, or health and drug monitoring reasons. Recently, metabonomics/metabolomics have emerged as a new and promising 'omics' platform that shows potential in biomarker discovery, especially in areas such as disease diagnosis, assessment of drug efficacy or toxicity. Blood is collected in various establishments in conditions that are not standardized. Next, the samples are prepared and analyzed using different methodologies or tools. When targeted analysis of key molecules (e.g., a drug or its metabolite[s]) is the aim, enforcement of certain measures or additional analyses may correct and harmonize these discrepancies. In omics fields such as those performed by holistic analytical approaches, no such rules or tools are available. As a result, comparison or correlation of results or data fusion becomes impractical. However, it becomes evident that such obstacles should be overcome in the near future to allow for large-scale studies that involve the assaying of samples from hundreds of individuals. In this case the effect of sample handling and preparation becomes very serious, in order to avoid wasting months of work from experts and expensive instrument time. The present review aims to cover the different methodologies applied to the pretreatment of blood prior to LC-MS metabolomic/metabonomic studies. The article tries to critically compare the methods and highlight issues that need to be addressed.

MIPs as Tools in Environmental Biotechnology.

PubMed

Mattiasson, Bo

2015-01-01

Molecular imprints are potentially fantastic constructions. They are selective, robust, and nonbiodegradable if produced from stable polymers. A range of different applications has been presented, everything from separation of enantiomers, via adsorbents for sample preparation before analysis to applications in wastewater treatment. This chapter deals with molecularly imprinted polymers (MIPs) as tools in environmental biotechnology, a field that has the potential to become very important in the future.

Solid-Phase Extraction (SPE): Principles and Applications in Food Samples.

PubMed

Ötles, Semih; Kartal, Canan

2016-01-01

Solid-Phase Extraction (SPE) is a sample preparation method that is practised on numerous application fields due to its many advantages compared to other traditional methods. SPE was invented as an alternative to liquid/liquid extraction and eliminated multiple disadvantages, such as usage of large amount of solvent, extended operation time/procedure steps, potential sources of error, and high cost. Moreover, SPE can be plied to the samples combined with other analytical methods and sample preparation techniques optionally. SPE technique is a useful tool for many purposes through its versatility. Isolation, concentration, purification and clean-up are the main approaches in the practices of this method. Food structures represent a complicated matrix and can be formed into different physical stages, such as solid, viscous or liquid. Therefore, sample preparation step particularly has an important role for the determination of specific compounds in foods. SPE offers many opportunities not only for analysis of a large diversity of food samples but also for optimization and advances. This review aims to provide a comprehensive overview on basic principles of SPE and its applications for many analytes in food matrix.

Quantitation of heat-shock proteins in clinical samples using mass spectrometry.

PubMed

Kaur, Punit; Asea, Alexzander

2011-01-01

Mass spectrometry (MS) is a powerful analytical tool for proteomics research and drug and biomarker discovery. MS enables identification and quantification of known and unknown compounds by revealing their structural and chemical properties. Proper sample preparation for MS-based analysis is a critical step in the proteomics workflow because the quality and reproducibility of sample extraction and preparation for downstream analysis significantly impact the separation and identification capabilities of mass spectrometers. The highly expressed proteins represent potential biomarkers that could aid in diagnosis, therapy, or drug development. Because the proteome is so complex, there is no one standard method for preparing protein samples for MS analysis. Protocols differ depending on the type of sample, source, experiment, and method of analysis. Molecular chaperones play significant roles in almost all biological functions due to their capacity for detecting intracellular denatured/unfolded proteins, initiating refolding or denaturation of such malfolded protein sequences and more recently for their role in the extracellular milieu as chaperokines. In this chapter, we describe the latest techniques for quantitating the expression of molecular chaperones in human clinical samples.

Size exclusion chromatography for analyses of fibroin in silk: optimization of sampling and separation conditions

NASA Astrophysics Data System (ADS)

Pawcenis, Dominika; Koperska, Monika A.; Milczarek, Jakub M.; Łojewski, Tomasz; Łojewska, Joanna

2014-02-01

A direct goal of this paper was to improve the methods of sample preparation and separation for analyses of fibroin polypeptide with the use of size exclusion chromatography (SEC). The motivation for the study arises from our interest in natural polymers included in historic textile and paper artifacts, and is a logical response to the urgent need for developing rationale-based methods for materials conservation. The first step is to develop a reliable analytical tool which would give insight into fibroin structure and its changes caused by both natural and artificial ageing. To investigate the influence of preparation conditions, two sets of artificially aged samples were prepared (with and without NaCl in sample solution) and measured by the means of SEC with multi angle laser light scattering detector. It was shown that dialysis of fibroin dissolved in LiBr solution allows removal of the salt which destroys stacks chromatographic columns and prevents reproducible analyses. Salt rich (NaCl) water solutions of fibroin improved the quality of chromatograms.

Preliminary Development of an Object-Oriented Optimization Tool

NASA Technical Reports Server (NTRS)

Pak, Chan-gi

2011-01-01

The National Aeronautics and Space Administration Dryden Flight Research Center has developed a FORTRAN-based object-oriented optimization (O3) tool that leverages existing tools and practices and allows easy integration and adoption of new state-of-the-art software. The object-oriented framework can integrate the analysis codes for multiple disciplines, as opposed to relying on one code to perform analysis for all disciplines. Optimization can thus take place within each discipline module, or in a loop between the central executive module and the discipline modules, or both. Six sample optimization problems are presented. The first four sample problems are based on simple mathematical equations; the fifth and sixth problems consider a three-bar truss, which is a classical example in structural synthesis. Instructions for preparing input data for the O3 tool are presented.

New tools for evaluating LQAS survey designs

PubMed Central

2014-01-01

Lot Quality Assurance Sampling (LQAS) surveys have become increasingly popular in global health care applications. Incorporating Bayesian ideas into LQAS survey design, such as using reasonable prior beliefs about the distribution of an indicator, can improve the selection of design parameters and decision rules. In this paper, a joint frequentist and Bayesian framework is proposed for evaluating LQAS classification accuracy and informing survey design parameters. Simple software tools are provided for calculating the positive and negative predictive value of a design with respect to an underlying coverage distribution and the selected design parameters. These tools are illustrated using a data example from two consecutive LQAS surveys measuring Oral Rehydration Solution (ORS) preparation. Using the survey tools, the dependence of classification accuracy on benchmark selection and the width of the ‘grey region’ are clarified in the context of ORS preparation across seven supervision areas. Following the completion of an LQAS survey, estimation of the distribution of coverage across areas facilitates quantifying classification accuracy and can help guide intervention decisions. PMID:24528928

New tools for evaluating LQAS survey designs.

PubMed

Hund, Lauren

2014-02-15

Lot Quality Assurance Sampling (LQAS) surveys have become increasingly popular in global health care applications. Incorporating Bayesian ideas into LQAS survey design, such as using reasonable prior beliefs about the distribution of an indicator, can improve the selection of design parameters and decision rules. In this paper, a joint frequentist and Bayesian framework is proposed for evaluating LQAS classification accuracy and informing survey design parameters. Simple software tools are provided for calculating the positive and negative predictive value of a design with respect to an underlying coverage distribution and the selected design parameters. These tools are illustrated using a data example from two consecutive LQAS surveys measuring Oral Rehydration Solution (ORS) preparation. Using the survey tools, the dependence of classification accuracy on benchmark selection and the width of the 'grey region' are clarified in the context of ORS preparation across seven supervision areas. Following the completion of an LQAS survey, estimation of the distribution of coverage across areas facilitates quantifying classification accuracy and can help guide intervention decisions.

Biomedical applications of laser-induced breakdown spectroscopy (LIBS)

NASA Astrophysics Data System (ADS)

Unnikrishnan, V. K.; Nayak, Rajesh; Bhat, Sujatha; Mathew, Stanley; Kartha, V. B.; Santhosh, C.

2015-03-01

LIBS has been proven to be a robust elemental analysis tool attracting interest because of the wide applications. LIBS can be used for analysis of any type of samples i.e. environmental/physiological, regardless of its state of matter. Conventional spectroscopy techniques are good in analytical performance, but their sample preparation method is mostly destructive and time consuming. Also, almost all these methods are incapable of analysing multi elements simaltaneously. On the other hand, LIBS has many potential advantages such as simplicity in the experimental setup, less sample preparation, less destructive analysis of sample etc. In this paper, we report some of the biomedical applications of LIBS. From the experiments carried out on clinical samples (calcified tissues or teeth and gall stones) for trace elemental mapping and detection, it was found that LIBS is a robust tool for such applications. It is seen that the presence and relative concentrations of major elements (calcium, phosphorus and magnesium) in human calcified tissue (tooth) can be easily determined using LIBS technique. The importance of this study comes in anthropology where tooth and bone are main samples from which reliable data can be easily retrieved. Similarly, elemental composition of bile juice and gall stone collected from the same subject using LIBS was found to be similar. The results show interesting prospects for LIBS to study cholelithiasis (the presence of stones in the gall bladder, is a common disease of the gastrointestinal tract) better.

Pre-Clinical Evaluation of a Real-Time PCR Assay on a Portable Instrument as a Possible Field Diagnostic Tool: Experiences from the Testing of Clinical Samples for African and Classical Swine Fever Viruses.

PubMed

Liu, L; Luo, Y; Accensi, F; Ganges, L; Rodríguez, F; Shan, H; Ståhl, K; Qiu, H-J; Belák, S

2017-10-01

African swine fever (ASF) and classical swine fever (CSF) are two highly infectious transboundary animal diseases (TADs) that are serious threats to the pig industry worldwide, including in China, the world's largest pork producer. In this study, a duplex real-time PCR assay was developed for the rapid detection and differentiation of African swine fever virus (ASFV) and classical swine fever virus (CSFV). The assay was performed on a portable, battery-powered PCR thermocycler with a low sample throughput (termed as 'T-COR4 assay'). The feasibility and reliability of the T-COR4 assay as a possible field method was investigated by testing clinical samples collected in China. When evaluated with reference materials or samples from experimental infections, the assay performed in a reliable manner, producing results comparable to those obtained from stationary PCR platforms. Of 59 clinical samples, 41 had results identical to a two-step CSFV real-time PCR assay. No ASFV was detected in these samples. The T-COR4 assay was technically easy to perform and produced results within 3 h, including sample preparation. In combination with a simple sample preparation method, the T-COR4 assay provides a new tool for the field diagnosis and differentiation of ASF and CSF, which could be of particular value in remote areas. © 2016 Blackwell Verlag GmbH.

Approaching Suspicious Substances Safely

NASA Technical Reports Server (NTRS)

2004-01-01

A mineral identification tool that was developed for NASA's Mars Rover Technology Development program is now serving as a powerful tool for U.S. law enforcement agencies and military personnel to identify suspicious liquid and solid substances. The tool can measure unknown substances through glass and plastic packaging materials with the RamanProbe(TradeMark) focused fiber-optic probe. The probe length can be extended up to 200 meters to enable users to analyze potentially dangerous substances at a safe distance. In many cases, the spectrometer and personnel are kept in a safe zone while the probe is positioned next to the sample being analyzed. Being able to identify chemicals in remote locations also saves users time and labor, since otherwise the samples would need to be collected, transported, and prepared prior to measurement in the laboratory.

Properties and Cutting Performance of TiAlSiN Coating Prepared by Cathode Arc Ion Plating

NASA Astrophysics Data System (ADS)

Zhang, Er-Geng; Chen, Qiang; Wang, Qin-Xue; Huang, Biao

2016-06-01

TiAlSiN coating was deposited on high-speed steel (HSS) samples and cemented carbide tool inserts, respectively, by a new coating preparation procedure, and its properties and cutting performance were characterized. The coating thickness, chemical composition, microstructure morphology and mechanical properties were investigated by X-ray fluorescence measurement system, energy dispersive spectrometer (EDS), scanning electron microscope (SEM), nanoindentation, Rockwell hardness tester and ball-on-disc tribometer. A 3D orthogonal cutting experiment model was established by DEFORM-3D to study the influences of different coating thicknesses on cutting force and temperature, and the field cutting experiment was carried out. The results show that the thickness of TiAlSiN coating is 3.14μm prepared by the 3μm preparation procedure, microhardness is 36.727GPa with the Si content of about 5.22at.% as well as good fracture toughness and adhesion strength. The TC4 and AISI 1045 cutting tool inserts with 4μm coating thickness have the minimum cutting forces of about 734.7N and 450.7N, respectively. Besides, tool inserts with a thickness of 3μm have the minimum cutting temperatures of about 510.2∘C and 230.6∘C, respectively.

The application of a novel optical SPM in biomedicine

NASA Astrophysics Data System (ADS)

Li, Yinli; Chen, Haibo; Wu, Shifa; Song, Linfeng; Zhang, Jian

2005-01-01

As an analysis tool, SPM has been broadly used in biomedicine in recent years, such as AFM and SNOM; they are effective instruments in detecting life nanostructures at atomic level. Atomic force and photon scanning tunneling microscope (AF/PSTM) is one of member of SPM, it can be used to obtain sample" optical and atomic fore images at once scanning, these images include the transmissivity image, reflection index image and topography image. This report mainly introduces the application of AF/PSTM in red blood membrane and the effect of different sample dealt with processes on the experiment result. The materials for preparing red cells membrane samples are anticoagulant blood, isotonic phosphatic buffer solution (PBS) and new two times distilled water. The images of AF/PSTM give real expression to the biology samples" fact despite of different sample dealt with processes, which prove that AF/PSTM suits to biology sample imaging. At the same time, the optical images and the topography image of AF/PSTM of the same sample are complementary with each other; this will make AF/PSTM a facile tool to analysis biologic samples" nanostructure. As another sample, this paper gives the application of AF/PSTM in immunoassay, the result shows that AF/PSTM is suit to analysis biologic sample, and it will become a new tool for biomedicine test.

MASS SPECTROMETRY-BASED METABOLOMICS

PubMed Central

Dettmer, Katja; Aronov, Pavel A.; Hammock, Bruce D.

2007-01-01

This review presents an overview of the dynamically developing field of mass spectrometry-based metabolomics. Metabolomics aims at the comprehensive and quantitative analysis of wide arrays of metabolites in biological samples. These numerous analytes have very diverse physico-chemical properties and occur at different abundance levels. Consequently, comprehensive metabolomics investigations are primarily a challenge for analytical chemistry and specifically mass spectrometry has vast potential as a tool for this type of investigation. Metabolomics require special approaches for sample preparation, separation, and mass spectrometric analysis. Current examples of those approaches are described in this review. It primarily focuses on metabolic fingerprinting, a technique that analyzes all detectable analytes in a given sample with subsequent classification of samples and identification of differentially expressed metabolites, which define the sample classes. To perform this complex task, data analysis tools, metabolite libraries, and databases are required. Therefore, recent advances in metabolomics bioinformatics are also discussed. PMID:16921475

Laboratory, Field, and Analytical Procedures for Using ...

EPA Pesticide Factsheets

Regardless of the remedial technology invoked to address contaminated sediments in the environment, there is a critical need to have tools for assessing the effectiveness of the remedy. In the past, these tools have included chemical and biomonitoring of the water column and sediments, toxicity testing and bioaccumulation studies performed on site sediments, and application of partitioning, transport and fate modeling. All of these tools served as lines of evidence for making informed environmental management decisions at contaminated sediment sites. In the last ten years, a new tool for assessing remedial effectiveness has gained a great deal of attention. Passive sampling offers a tool capable of measuring the freely dissolved concentration (Cfree) of legacy contaminants in water and sediments. In addition to assessing the effectiveness of the remedy, passive sampling can be applied for a variety of other contaminated sediments site purposes involved with performing the preliminary assessment and site inspection, conducting the remedial investigation and feasibility study, preparing the remedial design, and assessing the potential for contaminant bioaccumulation. While there is a distinct need for using passive sampling at contaminated sediments sites and several previous documents and research articles have discussed various aspects of passive sampling, there has not been definitive guidance on the laboratory, field and analytical procedures for using pas

Evaluation of the Illumigene Malaria LAMP: A Robust Molecular Diagnostic Tool for Malaria Parasites

PubMed Central

Lucchi, Naomi W.; Gaye, Marie; Diallo, Mammadou Alpha; Goldman, Ira F.; Ljolje, Dragan; Deme, Awa Bineta; Badiane, Aida; Ndiaye, Yaye Die; Barnwell, John W.; Udhayakumar, Venkatachalam; Ndiaye, Daouda

2016-01-01

Isothermal nucleic acid amplification assays such as the loop mediated isothermal amplification (LAMP), are well suited for field use as they do not require thermal cyclers to amplify the DNA. To further facilitate the use of LAMP assays in remote settings, simpler sample preparation methods and lyophilized reagents are required. The performance of a commercial malaria LAMP assay (Illumigene Malaria LAMP) was evaluated using two sample preparation workflows (simple filtration prep (SFP)) and gravity-driven filtration prep (GFP)) and pre-dispensed lyophilized reagents. Laboratory and clinical samples were tested in a field laboratory in Senegal and the results independently confirmed in a reference laboratory in the U.S.A. The Illumigene Malaria LAMP assay was easily implemented in the clinical laboratory and gave similar results to a real-time PCR reference test with limits of detection of ≤2.0 parasites/μl depending on the sample preparation method used. This assay reliably detected Plasmodium sp. parasites in a simple low-tech format, providing a much needed alternative to the more complex molecular tests for malaria diagnosis. PMID:27827432

Healthcare providers' attitudes and behaviours related to paediatric mild traumatic brain injury: results from the 2014 DocStyles survey.

PubMed

Sarmiento, Kelly; Donnell, Zoe; Hoffman, Rosanne; Tennant, Bethany

2018-04-23

Explore healthcare providers' experiences managing mTBI and better understand their use of mTBI assessment tools and guidelines. Cross-sectional Methods: A random sample of 1,760 healthcare providers responded to the web-based DocStyles survey between June 18 and 30, 2014. The sample included family/general practitioners, internists, pediatricians, and nurse practitioners who reported seeing pediatric patients. We examined their experiences with mTBI to identify opportunities to increase preparedness and improve management of mTBI. Fifty-nine percent of healthcare providers reported that they diagnosed or managed pediatric patients with mTBI within the last 12 months. Of those, 44.4% felt 'very prepared' to make decisions about when pediatric patients can safety return to activities, such as school and sports after a mTBI. When asked how often they use screening or assessment tools to assess pediatric patients with mTBI, almost half reported that they 'seldom' or 'never' use those resources (24.6% and 22.0%, respectively). Most healthcare providers reported seeing pediatric patients with mTBI, yet most feel only somewhat prepared to manage this injury in their practise. Broader use of screening tools and guidelines, that include clinical decision support tools, may be useful for healthcare providers who care for pediatric patients with mTBI.

Comparison of a prepCheck-supported self-assessment concept with conventional faculty supervision in a pre-clinical simulation environment.

PubMed

Wolgin, M; Grabowski, S; Elhadad, S; Frank, W; Kielbassa, A M

2018-03-25

This study aimed to evaluate the educational outcome of a digitally based self-assessment concept (prepCheck; DentsplySirona, Wals, Austria) for pre-clinical undergraduates in the context of a regular phantom-laboratory course. A sample of 47 third-year dental students participated in the course. Students were randomly divided into a prepCheck-supervised (self-assessment) intervention group (IG; n = 24); conventionally supervised students constituted the control group (CG; n = 23). During the preparation of three-surface (MOD) class II amalgam cavities, each IG participant could analyse a superimposed 3D image of his/her preparation against the "master preparation" using the prepCheck software. In the CG, several course instructors performed the evaluations according to pre-defined assessment criteria. After completing the course, a mandatory (blinded) practical examination was taken by all course participants (both IG and CG students), and this assessment involved the preparation of a MOD amalgam cavity. Then, optical impressions by means of a CEREC-Omnicam were taken to digitalize all examination preparations, followed by surveying and assessing the latter using prepCheck. The statistical analysis of the digitalized samples (Mann-Whitney U test) revealed no significant differences between the cavity dimensions achieved in the IG and CG (P = .406). Additionally, the sum score of the degree of conformity with the "master preparation" (maximum permissible 10% of plus or minus deviation) was comparable in both groups (P = .259). The implemented interactive digitally based, self-assessment learning tool for undergraduates appears to be equivalent to the conventional form of supervision. Therefore, such digital learning tools could significantly address the ever-increasing student to faculty ratio. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography.

PubMed

Trépout, Sylvain; Bastin, Philippe; Marco, Sergio

2017-03-12

This report describes a protocol for preparing thick biological specimens for further observation using a scanning transmission electron microscope. It also describes an imaging method for studying the 3D structure of thick biological specimens by scanning transmission electron tomography. The sample preparation protocol is based on conventional methods in which the sample is fixed using chemical agents, treated with a heavy atom salt contrasting agent, dehydrated in a series of ethanol baths, and embedded in resin. The specific imaging conditions for observing thick samples by scanning transmission electron microscopy are then described. Sections of the sample are observed using a through-focus method involving the collection of several images at various focal planes. This enables the recovery of in-focus information at various heights throughout the sample. This particular collection pattern is performed at each tilt angle during tomography data collection. A single image is then generated, merging the in-focus information from all the different focal planes. A classic tilt-series dataset is then generated. The advantage of the method is that the tilt-series alignment and reconstruction can be performed using standard tools. The collection of through-focal images allows the reconstruction of a 3D volume that contains all of the structural details of the sample in focus.

Thermoluminescence glow curve analysis and CGCD method for erbium doped CaZrO{sub 3} phosphor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tiwari, Ratnesh, E-mail: 31rati@gmail.com; Chopra, Seema

2016-05-06

The manuscript report the synthesis, thermoluminescence study at fixed concentration of Er{sup 3+} (1 mol%) doped CaZrO{sub 3} phosphor. The phosphors were prepared by modified solid state reaction method. The powder sample was characterized by thermoluminescence (TL) glow curve analysis. In TL glow curve the optimized concentration in 1mol% for UV irradiated sample. The kinetic parameters were calculated by computerized glow curve deconvolution (CGCD) techniaue. Trapping parameters gives the information of dosimetry loss in prepared phosphor and its usability in environmental monitoring and for personal monitoring. CGCD is the advance tool for analysis of complicated TL glow curves.

SMART: A Propositional Logic-Based Trade Analysis and Risk Assessment Tool for a Complex Mission

NASA Technical Reports Server (NTRS)

Ono, Masahiro; Nicholas, Austin; Alibay, Farah; Parrish, Joseph

2015-01-01

This paper introduces a new trade analysis software called the Space Mission Architecture and Risk Analysis Tool (SMART). This tool supports a high-level system trade study on a complex mission, such as a potential Mars Sample Return (MSR) mission, in an intuitive and quantitative manner. In a complex mission, a common approach to increase the probability of success is to have redundancy and prepare backups. Quantitatively evaluating the utility of adding redundancy to a system is important but not straightforward, particularly when the failure of parallel subsystems are correlated.

Pressure-Assisted Chelating Extraction as a Teaching Tool in Instrumental Analysis

ERIC Educational Resources Information Center

Sadik, Omowunmi A.; Wanekaya, Adam K.; Yevgeny, Gelfand

2004-01-01

A novel instrumental-digestion technique using pressure-assisted chelating extraction (PACE), for undergraduate laboratory is reported. This procedure is used for exposing students to safe sample-preparation techniques, for correlating wet-chemical methods with modern instrumental analysis and comparing the performance of PACE with conventional…

Extension of an Object-Oriented Optimization Tool: User's Reference Manual

NASA Technical Reports Server (NTRS)

Pak, Chan-Gi; Truong, Samson S.

2015-01-01

The National Aeronautics and Space Administration Armstrong Flight Research Center has developed a cost-effective and flexible object-oriented optimization (O (sup 3)) tool that leverages existing tools and practices and allows easy integration and adoption of new state-of-the-art software. This object-oriented framework can integrate the analysis codes for multiple disciplines, as opposed to relying on one code to perform analysis for all disciplines. Optimization can thus take place within each discipline module, or in a loop between the O (sup 3) tool and the discipline modules, or both. Six different sample mathematical problems are presented to demonstrate the performance of the O (sup 3) tool. Instructions for preparing input data for the O (sup 3) tool are detailed in this user's manual.

FISH-in-CHIPS: A Microfluidic Platform for Molecular Typing of Cancer Cells.

PubMed

Perez-Toralla, Karla; Mottet, Guillaume; Tulukcuoglu-Guneri, Ezgi; Champ, Jérôme; Bidard, François-Clément; Pierga, Jean-Yves; Klijanienko, Jerzy; Draskovic, Irena; Malaquin, Laurent; Viovy, Jean-Louis; Descroix, Stéphanie

2017-01-01

Microfluidics offer powerful tools for the control, manipulation, and analysis of cells, in particular for the assessment of cell malignancy or the study of cell subpopulations. However, implementing complex biological protocols on chip remains a challenge. Sample preparation is often performed off chip using multiple manually performed steps, and protocols usually include different dehydration and drying steps that are not always compatible with a microfluidic format.Here, we report the implementation of a Fluorescence in situ Hybridization (FISH) protocol for the molecular typing of cancer cells in a simple and low-cost device. The geometry of the chip allows integrating the sample preparation steps to efficiently assess the genomic content of individual cells using a minute amount of sample. The FISH protocol can be fully automated, thus enabling its use in routine clinical practice.

Liquid Phase Sintering

NASA Technical Reports Server (NTRS)

2004-01-01

Industry spends billions of dollars each year on machine tools to manufacture products out of metal. This includes tools for cutting every kind of metal part from engine blocks to Shuttle main engine components. Cutting tool tips often break because of weak spots or defects in their composition. Based on a new concept called defect trapping, space offers a novel environment to study defect formation in molten metal materials as they solidify. After the return of these materials from space, researchers can evaluate the source of the defect and seek ways to eliminate them in products prepared on Earth. A widely used process for cutting tip manufacturing is liquid phase sintering. Compared to Earth-sintered samples which slump due to buoyancy induced by gravity, space samples are uniformly shaped and defects remain where they are formed. By studying metals sintered in space the US tool industry can potentially enhance its worldwide competitiveness. The Consortium for Materials Development in Space along with Wyle Labs, Teledyne Advanced Materials, and McDornell Douglas have conducted experiments in space.

Microgravity

NASA Image and Video Library

2004-04-15

Industry spends billions of dollars each year on machine tools to manufacture products out of metal. This includes tools for cutting every kind of metal part from engine blocks to Shuttle main engine components. Cutting tool tips often break because of weak spots or defects in their composition. Based on a new concept called defect trapping, space offers a novel environment to study defect formation in molten metal materials as they solidify. After the return of these materials from space, researchers can evaluate the source of the defect and seek ways to eliminate them in products prepared on Earth. A widely used process for cutting tip manufacturing is liquid phase sintering. Compared to Earth-sintered samples which slump due to buoyancy induced by gravity, space samples are uniformly shaped and defects remain where they are formed. By studying metals sintered in space the US tool industry can potentially enhance its worldwide competitiveness. The Consortium for Materials Development in Space along with Wyle Labs, Teledyne Advanced Materials, and McDornell Douglas have conducted experiments in space.

Can We Prepare Effective Writing Teachers for Academically Diverse Classrooms?

ERIC Educational Resources Information Center

Uyar, Yusuf

2016-01-01

This study examines whether preservice teachers, who provide writing instruction in native language education, are ready to teach students with different learning needs effectively. This study used a survey research design and qualitative data collection tools. An interview form and writing samples from students in the same class with different…

Imaging MALDI MS of Dosed Brain Tissues Utilizing an Alternative Analyte Pre-extraction Approach

NASA Astrophysics Data System (ADS)

Quiason, Cristine M.; Shahidi-Latham, Sheerin K.

2015-06-01

Matrix-assisted laser desorption ionization (MALDI) imaging mass spectrometry has been adopted in the pharmaceutical industry as a useful tool to detect xenobiotic distribution within tissues. A unique sample preparation approach for MALDI imaging has been described here for the extraction and detection of cobimetinib and clozapine, which were previously undetectable in mouse and rat brain using a single matrix application step. Employing a combination of a buffer wash and a cyclohexane pre-extraction step prior to standard matrix application, the xenobiotics were successfully extracted and detected with an 8 to 20-fold gain in sensitivity. This alternative approach for sample preparation could serve as an advantageous option when encountering difficult to detect analytes.

Cancer diagnosis by infrared spectroscopy: methodological aspects

NASA Astrophysics Data System (ADS)

Jackson, Michael; Kim, Keith; Tetteh, John; Mansfield, James R.; Dolenko, Brion; Somorjai, Raymond L.; Orr, F. W.; Watson, Peter H.; Mantsch, Henry H.

1998-04-01

IR spectroscopy is proving to be a powerful tool for the study and diagnosis of cancer. The application of IR spectroscopy to the analysis of cultured tumor cells and grading of breast cancer sections is outlined. Potential sources of error in spectral interpretation due to variations in sample histology and artifacts associated with sample storage and preparation are discussed. The application of statistical techniques to assess differences between spectra and to non-subjectively classify spectra is demonstrated.

Utility of eButton images for identifying food preparation behaviors and meal-related tasks in adolescents.

PubMed

Raber, Margaret; Patterson, Monika; Jia, Wenyan; Sun, Mingui; Baranowski, Tom

2018-02-24

Food preparation skills may encourage healthy eating. Traditional assessment of child food preparation employs self- or parent proxy-reporting methods, which are prone to error. The eButton is a wearable all-day camera that has promise as an objective, passive method for measuring child food preparation practices. This paper explores the feasibility of the eButton to reliably capture home food preparation behaviors and practices in a sample of pre- and early adolescents (ages 9 to 13). This is a secondary analysis of two eButton pilot projects evaluating the dietary intake of pre- and early adolescents in or around Houston, Texas. Food preparation behaviors were coded into seven major categories including: browsing, altering food/adding seasoning, food media, meal related tasks, prep work, cooking and observing. Inter-coder reliability was measured using Cohen's kappa and percent agreement. Analysis was completed on data for 31 participants. The most common activity was browsing in the pantry or fridge. Few participants demonstrated any food preparation work beyond unwrapping of food packages and combining two or more ingredients; actual cutting or measuring of foods were rare. Although previous research suggests children who "help" prepare meals may obtain some dietary benefit, accurate assessment tools of food preparation behavior are lacking. The eButton offers a feasible approach to food preparation behavior measurement among pre- and early adolescents. Follow up research exploring the validity of this method in a larger sample, and comparisons between cooking behavior and dietary intake are needed.

Direct Blood Dry LAMP: A Rapid, Stable, and Easy Diagnostic Tool for Human African Trypanosomiasis

PubMed Central

Hayashida, Kyoko; Kajino, Kiichi; Hachaambwa, Lottie; Namangala, Boniface; Sugimoto, Chihiro

2015-01-01

Loop-mediated isothermal amplification (LAMP) is a rapid and sensitive tool used for the diagnosis of a variety of infectious diseases. One of the advantages of this method over the polymerase chain reaction is that DNA amplification occurs at a constant temperature, usually between 60–65°C; therefore, expensive devices are unnecessary for this step. However, LAMP still requires complicated sample preparation steps and a well-equipped laboratory to produce reliable and reproducible results, which limits its use in resource-poor laboratories in most developing countries. In this study, we made several substantial modifications to the technique to carry out on-site diagnosis of Human African Trypanosomiasis (HAT) in remote areas using LAMP. The first essential improvement was that LAMP reagents were dried and stabilized in a single tube by incorporating trehalose as a cryoprotectant to prolong shelf life at ambient temperature. The second technical improvement was achieved by simplifying the sample preparation step so that DNA or RNA could be amplified directly from detergent-lysed blood samples. With these modifications, diagnosis of HAT in local clinics or villages in endemic areas becomes a reality, which could greatly impact on the application of diagnosis not only for HAT but also for other tropical diseases. PMID:25769046

Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters

PubMed Central

Holling, Nina; Dedi, Cinzia; Jones, Caroline E; Hawthorne, Joseph A; Hanlon, Geoffrey W; Salvage, Jonathan P; Patel, Bhavik A; Barnes, Lara M; Jones, Brian V

2014-01-01

Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation. PMID:24786314

Multiscale dispersion-state characterization of nanocomposites using optical coherence tomography

PubMed Central

Schneider, Simon; Eppler, Florian; Weber, Marco; Olowojoba, Ganiu; Weiss, Patrick; Hübner, Christof; Mikonsaari, Irma; Freude, Wolfgang; Koos, Christian

2016-01-01

Nanocomposite materials represent a success story of nanotechnology. However, development of nanomaterial fabrication still suffers from the lack of adequate analysis tools. In particular, achieving and maintaining well-dispersed particle distributions is a key challenge, both in material development and industrial production. Conventional methods like optical or electron microscopy need laborious, costly sample preparation and do not permit fast extraction of nanoscale structural information from statistically relevant sample volumes. Here we show that optical coherence tomography (OCT) represents a versatile tool for nanomaterial characterization, both in a laboratory and in a production environment. The technique does not require sample preparation and is applicable to a wide range of solid and liquid material systems. Large particle agglomerates can be directly found by OCT imaging, whereas dispersed nanoparticles are detected by model-based analysis of depth-dependent backscattering. Using a model system of polystyrene nanoparticles, we demonstrate nanoparticle sizing with high accuracy. We further prove the viability of the approach by characterizing highly relevant material systems based on nanoclays or carbon nanotubes. The technique is perfectly suited for in-line metrology in a production environment, which is demonstrated using a state-of-the-art compounding extruder. These experiments represent the first demonstration of multiscale nanomaterial characterization using OCT. PMID:27557544

Multiscale dispersion-state characterization of nanocomposites using optical coherence tomography.

PubMed

Schneider, Simon; Eppler, Florian; Weber, Marco; Olowojoba, Ganiu; Weiss, Patrick; Hübner, Christof; Mikonsaari, Irma; Freude, Wolfgang; Koos, Christian

2016-08-25

Nanocomposite materials represent a success story of nanotechnology. However, development of nanomaterial fabrication still suffers from the lack of adequate analysis tools. In particular, achieving and maintaining well-dispersed particle distributions is a key challenge, both in material development and industrial production. Conventional methods like optical or electron microscopy need laborious, costly sample preparation and do not permit fast extraction of nanoscale structural information from statistically relevant sample volumes. Here we show that optical coherence tomography (OCT) represents a versatile tool for nanomaterial characterization, both in a laboratory and in a production environment. The technique does not require sample preparation and is applicable to a wide range of solid and liquid material systems. Large particle agglomerates can be directly found by OCT imaging, whereas dispersed nanoparticles are detected by model-based analysis of depth-dependent backscattering. Using a model system of polystyrene nanoparticles, we demonstrate nanoparticle sizing with high accuracy. We further prove the viability of the approach by characterizing highly relevant material systems based on nanoclays or carbon nanotubes. The technique is perfectly suited for in-line metrology in a production environment, which is demonstrated using a state-of-the-art compounding extruder. These experiments represent the first demonstration of multiscale nanomaterial characterization using OCT.

A novel method for preparing histology slides to integrate the teaching of gross and microscopic anatomy.

PubMed

Provo-Klimek, Judy A; Troyer, Deryl L

2002-01-01

The authors have previously reported the development of a novel technique for sampling and preparing tissue slides for routine microscopic examination, without the use of a microtome. Termed "RAMP" (Rapid Adhesive Mediated Procedure), this simple, albeit somewhat crude, technique holds promise as a method that can be used in the field by veterinary practitioners for rapid microscopic evaluations to obtain early preliminary estimates of the nature of a mass or lesion. We incorporated the use of this method into a gross anatomy course in an attempt to gauge its utility for novices in tissue sampling and histology slide preparation. By having each group of students take a tissue sample from their cadaver, the activity simulated an actual necropsy situation in which practitioners in the field might use the technique. Because students were able to follow their specimen from sampling to microscopic examination, the activity provided a valuable integration of their learning of gross and microscopic anatomy. We conducted an evaluation of the process and the resulting slides with two successive classes of students. We conclude that the RAMP method is reasonably successful in the hands of individuals not trained in tissue preparation; was well received by the students as a valuable learning tool; and could potentially yield useful histological information for practicing veterinarians. Limitations of the method are also discussed.

Next-generation sequencing library preparation method for identification of RNA viruses on the Ion Torrent Sequencing Platform.

PubMed

Chen, Guiqian; Qiu, Yuan; Zhuang, Qingye; Wang, Suchun; Wang, Tong; Chen, Jiming; Wang, Kaicheng

2018-05-09

Next generation sequencing (NGS) is a powerful tool for the characterization, discovery, and molecular identification of RNA viruses. There were multiple NGS library preparation methods published for strand-specific RNA-seq, but some methods are not suitable for identifying and characterizing RNA viruses. In this study, we report a NGS library preparation method to identify RNA viruses using the Ion Torrent PGM platform. The NGS sequencing adapters were directly inserted into the sequencing library through reverse transcription and polymerase chain reaction, without fragmentation and ligation of nucleic acids. The results show that this method is simple to perform, able to identify multiple species of RNA viruses in clinical samples.

Application of refractometry to quality assurance monitoring of parenteral nutrition solutions.

PubMed

Chang, Wei-Kuo; Chao, You-Chen; Yeh, Ming-Kung

2008-01-01

Parenteral nutrition (PN) solution contains various concentrations of dextrose, amino acids, lipids, vitamins, electrolytes, and trace elements. Incorrect preparation of PN solution could lead to patient death. In this study we used the refractive index as a quality assurance tool to monitor the preparation of PN solution. Refractive indices of single nutrient components and PN solutions consisting of various concentrations of dextrose, amino acids, electrolytes, and lipids were measured. A mathematical equation and its linear plot were generated then used to predict the refractive index of the PN solution. The best-fit refractive index for PN solution (i.e., the predicted refractive index)=0.9798x(% dextrose)+1.2889x(% amino acids)+1.1017x(% lipids)+0.9440x(% sum of the electrolytes)+0.5367 (r2=0.99). This equation was validated by comparing the measured refractive indices of 500 clinical PN solutions to their predicted refractive indices. We found that 2 of the 500 prepared samples (0.4%) had less than the predicted refractive index (<95%). Refractive index can be used as a reliable quality assurance tool for monitoring PN preparation. Such information can be obtained at the bedside and used to confirm the accuracy of the PN solution composition.

Preparation and Characterization of PETI-330/Multiwalled Carbon Nanotube

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Working, Dennis C.; Criss, Jim M.; Siochi, Emilie J.; Connell, John W.

2005-01-01

As part of an ongoing effort to incorporate multifunctionality into advanced composites, blends of PETI-330 and multi-walled carbon nanotubes (MWCNTs) were prepared, characterized and fabricated into moldings. The PETI-330/MWCNT mixtures were prepared at concentrations ranging from 3 to 25 weight percent by dry mixing the components in a ball mill. The resulting powders were characterized for degree of mixing, thermal and rheological properties. Based on the characterization results, PETI-330/MWCNT samples were scaled up to 300 g and used to fabricate moldings 10.2 cm x 15.2 cm x 0.32 cm thick. The moldings were made by injecting the mixtures at 260-280 C into an Invar tool followed by curing for 1 h at 371 C. The tool was designed to impart shear during the injection process in an attempt to achieve some alignment of the MWCNTs in the flow direction. Good quality moldings were obtained that were subsequently characterized for thermal, mechanical and electrical properties. The degree of dispersion and alignment of the MWCNTs were investigated using high-resolution scanning electron microscopy. The preparation and preliminary characterization of PETI-330/MWCNT composites will be discussed. Keywords: phenylethynyl terminated imides, high temperature polymers, nanocomposites, moldings

Perceived Utility of Pharmacy Licensure Examination Preparation Tools

PubMed Central

Peak, Amy Sutton; Sheehan, Amy Heck; Arnett, Stephanie

2006-01-01

Objectives To identify board examination preparation tools most commonly used by recent pharmacy graduates and determine which tools are perceived as most valuable and representative of the actual content of licensure examinations. Methods An electronic survey was sent to all 2004 graduates of colleges of pharmacy in Indiana. Participants identified which specific preparation tools were used and rated tools based on usefulness, representativeness of licensure examination, and monetary value, and provided overall recommendations to future graduates. Results The most commonly used preparation tools were the Pharmacy Law Review Session offered by Dr. Thomas Wilson at Purdue University, the Complete Review for Pharmacy, Pre-NAPLEX, PharmPrep, and the Kaplan NAPLEX Review. Tools receiving high ratings in all categories included Dr. Wilson's Pharmacy Law Review Session, Pre-NAPLEX, Comprehensive Pharmacy Review, Kaplan NAPLEX Review, and Review of Pharmacy. Conclusions Although no preparation tool was associated with a higher examination pass rate, certain tools were clearly rated higher than others by test takers. PMID:17149406

Current Technical Approaches for the Early Detection of Foodborne Pathogens: Challenges and Opportunities.

PubMed

Cho, Il-Hoon; Ku, Seockmo

2017-09-30

The development of novel and high-tech solutions for rapid, accurate, and non-laborious microbial detection methods is imperative to improve the global food supply. Such solutions have begun to address the need for microbial detection that is faster and more sensitive than existing methodologies (e.g., classic culture enrichment methods). Multiple reviews report the technical functions and structures of conventional microbial detection tools. These tools, used to detect pathogens in food and food homogenates, were designed via qualitative analysis methods. The inherent disadvantage of these analytical methods is the necessity for specimen preparation, which is a time-consuming process. While some literature describes the challenges and opportunities to overcome the technical issues related to food industry legal guidelines, there is a lack of reviews of the current trials to overcome technological limitations related to sample preparation and microbial detection via nano and micro technologies. In this review, we primarily explore current analytical technologies, including metallic and magnetic nanomaterials, optics, electrochemistry, and spectroscopy. These techniques rely on the early detection of pathogens via enhanced analytical sensitivity and specificity. In order to introduce the potential combination and comparative analysis of various advanced methods, we also reference a novel sample preparation protocol that uses microbial concentration and recovery technologies. This technology has the potential to expedite the pre-enrichment step that precedes the detection process.

AQME: A forensic mitochondrial DNA analysis tool for next-generation sequencing data.

PubMed

Sturk-Andreaggi, Kimberly; Peck, Michelle A; Boysen, Cecilie; Dekker, Patrick; McMahon, Timothy P; Marshall, Charla K

2017-11-01

The feasibility of generating mitochondrial DNA (mtDNA) data has expanded considerably with the advent of next-generation sequencing (NGS), specifically in the generation of entire mtDNA genome (mitogenome) sequences. However, the analysis of these data has emerged as the greatest challenge to implementation in forensics. To address this need, a custom toolkit for use in the CLC Genomics Workbench (QIAGEN, Hilden, Germany) was developed through a collaborative effort between the Armed Forces Medical Examiner System - Armed Forces DNA Identification Laboratory (AFMES-AFDIL) and QIAGEN Bioinformatics. The AFDIL-QIAGEN mtDNA Expert, or AQME, generates an editable mtDNA profile that employs forensic conventions and includes the interpretation range required for mtDNA data reporting. AQME also integrates an mtDNA haplogroup estimate into the analysis workflow, which provides the analyst with phylogenetic nomenclature guidance and a profile quality check without the use of an external tool. Supplemental AQME outputs such as nucleotide-per-position metrics, configurable export files, and an audit trail are produced to assist the analyst during review. AQME is applied to standard CLC outputs and thus can be incorporated into any mtDNA bioinformatics pipeline within CLC regardless of sample type, library preparation or NGS platform. An evaluation of AQME was performed to demonstrate its functionality and reliability for the analysis of mitogenome NGS data. The study analyzed Illumina mitogenome data from 21 samples (including associated controls) of varying quality and sample preparations with the AQME toolkit. A total of 211 tool edits were automatically applied to 130 of the 698 total variants reported in an effort to adhere to forensic nomenclature. Although additional manual edits were required for three samples, supplemental tools such as mtDNA haplogroup estimation assisted in identifying and guiding these necessary modifications to the AQME-generated profile. Along with profile generation, AQME reported accurate haplogroups for 18 of the 19 samples analyzed. The single errant haplogroup assignment, although phylogenetically close, identified a bug that only affects partial mitogenome data. Future adjustments to AQME's haplogrouping tool will address this bug as well as enhance the overall scoring strategy to better refine and automate haplogroup assignments. As NGS enables broader use of the mtDNA locus in forensics, the availability of AQME and other forensic-focused mtDNA analysis tools will ease the transition and further support mitogenome analysis within routine casework. Toward this end, the AFMES-AFDIL has utilized the AQME toolbox in conjunction with the CLC Genomics Workbench to successfully validate and implement two NGS mitogenome methods. Copyright © 2017 Elsevier B.V. All rights reserved.

Rapid microscale in-gel processing and digestion of proteins using surface acoustic waves.

PubMed

Kulkarni, Ketav P; Ramarathinam, Sri H; Friend, James; Yeo, Leslie; Purcell, Anthony W; Perlmutter, Patrick

2010-06-21

A new method for in-gel sample processing and tryptic digestion of proteins is described. Sample preparation, rehydration, in situ digestion and peptide extraction from gel slices are dramatically accelerated by treating the gel slice with surface acoustic waves (SAWs). Only 30 minutes total workflow time is required for this new method to produce base peak chromatograms (BPCs) of similar coverage and intensity to those observed for traditional processing and overnight digestion. Simple set up, good reproducibility, excellent peptide recoveries, rapid turnover of samples and high confidence protein identifications put this technology at the fore-front of the next generation of proteomics sample processing tools.

Microextraction by Packed Sorbent (MEPS) and Solid-Phase Microextraction (SPME) as Sample Preparation Procedures for the Metabolomic Profiling of Urine

PubMed Central

Silva, Catarina; Cavaco, Carina; Perestrelo, Rosa; Pereira, Jorge; Câmara, José S.

2014-01-01

For a long time, sample preparation was unrecognized as a critical issue in the analytical methodology, thus limiting the performance that could be achieved. However, the improvement of microextraction techniques, particularly microextraction by packed sorbent (MEPS) and solid-phase microextraction (SPME), completely modified this scenario by introducing unprecedented control over this process. Urine is a biological fluid that is very interesting for metabolomics studies, allowing human health and disease characterization in a minimally invasive form. In this manuscript, we will critically review the most relevant and promising works in this field, highlighting how the metabolomic profiling of urine can be an extremely valuable tool for the early diagnosis of highly prevalent diseases, such as cardiovascular, oncologic and neurodegenerative ones. PMID:24958388

Performance Evaluation Test of the Rapid Area Preparation Tool (RAPTOR)

DTIC Science & Technology

2008-12-01

the standard SETCO tires. A blast test of the new SETCO tire is scheduled for the spring of...Washington, DC 20301-2500 Performance Evaluation Test of the Rapid Area Preparation Tool (RAPTOR) December 2008 Prepared ...2008 to 00-00-2008 4. TITLE AND SUBTITLE Performance Evaluation Test of the Rapid Area Preparation Tool (RAPTOR) 5a. CONTRACT NUMBER 5b.

High Temperature Resin/Carbon Nanotube Composite Fabrication

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Sun, Keun J.; Criss, Jim M.; Siochi, Emilie J.; Connell, John W.

2006-01-01

For the purpose of incorporating multifunctionality into advanced composites, blends of phenylethynyl terminated imides-330 (PETI-330) and multi-walled carbon nanotubes (MWCNTs) were prepared, characterized and fabricated into moldings. PETI-330/MWCNT mixtures were prepared at concentrations ranging from 3 to 25 weight percent by dry mixing the components in a ball mill. The resulting powders were characterized for degree of mixing, thermal and rheological properties. Based on the characterization results, PETI-330/MWCNT samples were scaled up to approximately 300 g and used to fabricate moldings by injecting the mixtures at 260-280 deg C into a stainless steel tool followed by curing for 1 h at 371 deg C. The tool was designed to impart a degree of shear during the injection process in an attempt to achieve some alignment of the MWCNTs in the flow direction. Obtained moldings were subsequently characterized for thermal, mechanical, and electrical properties. The degree of dispersion and alignment of MWCNTs were investigated using high-resolution scanning electron microscopy. The preparation and preliminary characterization of PETI-330/MWCNT composites will be discussed.

Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation.

PubMed

Ashtiani, Dariush; Venugopal, Hari; Belousoff, Matthew; Spicer, Bradley; Mak, Johnson; Neild, Adrian; de Marco, Alex

2018-04-06

Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6-8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves. Copyright © 2018 Elsevier Inc. All rights reserved.

Experimental investigation and optimization of welding process parameters for various steel grades using NN tool and Taguchi method

NASA Astrophysics Data System (ADS)

Soni, Sourabh Kumar; Thomas, Benedict

2018-04-01

The term "weldability" has been used to describe a wide variety of characteristics when a material is subjected to welding. In our analysis we perform experimental investigation to estimate the tensile strength of welded joint strength and then optimization of welding process parameters by using taguchi method and Artificial Neural Network (ANN) tool in MINITAB and MATLAB software respectively. The study reveals the influence on weldability of steel by varying composition of steel by mechanical characterization. At first we prepare the samples of different grades of steel (EN8, EN 19, EN 24). The samples were welded together by metal inert gas welding process and then tensile testing on Universal testing machine (UTM) was conducted for the same to evaluate the tensile strength of the welded steel specimens. Further comparative study was performed to find the effects of welding parameter on quality of weld strength by employing Taguchi method and Neural Network tool. Finally we concluded that taguchi method and Neural Network Tool is much efficient technique for optimization.

Chemical and Biological Tools for the Preparation of Modified Histone Proteins

PubMed Central

Howard, Cecil J.; Yu, Ruixuan R.; Gardner, Miranda L.; Shimko, John C.; Ottesen, Jennifer J.

2016-01-01

Eukaryotic chromatin is a complex and dynamic system in which the DNA double helix is organized and protected by interactions with histone proteins. This system is regulated through, a large network of dynamic post-translational modifications (PTMs) exists to ensure proper gene transcription, DNA repair, and other processes involving DNA. Homogenous protein samples with precisely characterized modification sites are necessary to better understand the functions of modified histone proteins. Here, we discuss sets of chemical and biological tools that have been developed for the preparation of modified histones, with a focus on the appropriate choice of tool for a given target. We start with genetic approaches for the creation of modified histones, including the incorporation of genetic mimics of histone modifications, chemical installation of modification analogs, and the use of the expanded genetic code to incorporate modified amino acids. Additionally, we will cover the chemical ligation techniques that have been invaluable in the generation of complex modified histones that are indistinguishable from the natural counterparts. Finally, we will end with a prospectus on future directions of synthetic chromatin in living systems. PMID:25863817

Semiautomated Sample Preparation for Protein Stability and Formulation Screening via Buffer Exchange.

PubMed

Ying, William; Levons, Jaquan K; Carney, Andrea; Gandhi, Rajesh; Vydra, Vicky; Rubin, A Erik

2016-06-01

A novel semiautomated buffer exchange process workflow was developed to enable efficient early protein formulation screening. An antibody fragment protein, BMSdab, was used to demonstrate the workflow. The process afforded 60% to 80% cycle time and scientist time savings and significant material efficiencies. These efficiencies ultimately facilitated execution of this stability work earlier in the drug development process, allowing this tool to inform the developability of potential candidates for development from a formulation perspective. To overcome the key technical challenges, the protein solution was buffer-exchanged by centrifuge filtration into formulations for stability screening in a 96-well plate with an ultrafiltration membrane, leveraging automated liquid handling and acoustic volume measurements to allow several cycles of exchanges. The formulations were transferred into a vacuum manifold and sterile filtered into a rack holding 96 glass vials. The vials were sealed with a capmat of individual caps and placed in stability stations. Stability of the samples prepared by this process and by the standard process was demonstrated to be comparable. This process enabled screening a number of formulations of a protein at an early pharmaceutical development stage with a short sample preparation time. © 2015 Society for Laboratory Automation and Screening.

Validated HPLC Determination of 4-Dimethylaminoantipyrine in Different Suppository Bases

PubMed Central

Kalmár, É; Kormányos, B.; Szakonyi, G.; Dombi, G.

2014-01-01

Suppositories are important tools for individual therapy, especially in paediatrics, and an instrumental assay method has become necessary for the quality control of dosage units. The aim of this work was to develop a rapid, effective high-performance liquid chromatography method to assay aminophenazone in extemporaneous suppositories prepared with two different suppository bases, adeps solidus and massa macrogoli. With a novel sample preparation method developed by the authors, 4-dimethylaminoantipyrine was determined in these suppository bases with 95-105% recovery. The measurements were carried out on a Shimadzu Prominence ultra high-performance liquid chromatography system equipped with a 20 μl sample loop. The separation was achieved on a Hypersil ODS column, with methanol, sodium acetate buffer (pH 5.5±0.05, 0.05 M, 60:40, v/v) as the mobile phase at a flow rate of 1.5 ml/min. The chromatograms were acquired at 253 nm. The chromatographic method was fully validated in accordance with current guidelines. The presented data demonstrate the successful development of a rapid, efficient and robust sample preparation and high-performance liquid chromatography method for the routine quality control of the dosage units of suppositories containing 4-dimethylaminoantipyrine. PMID:24799736

Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters.

PubMed

Holling, Nina; Dedi, Cinzia; Jones, Caroline E; Hawthorne, Joseph A; Hanlon, Geoffrey W; Salvage, Jonathan P; Patel, Bhavik A; Barnes, Lara M; Jones, Brian V

2014-06-01

Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation. © 2014 The Authors. FEMS Microbiology Letters published by John Wiley & Sons Ltd on behalf of Federation of European Microbiological Societies.

Efficacy of a geriatric oral health CD as a learning tool.

PubMed

Teasdale, Thomas A; Shaikh, Mehtab

2006-12-01

To better prepare professionals to meet the needs of older patients, a self-instructional computer module on geriatric oral health was previously developed. A follow-up study reported here tested the efficacy of this educational tool for improving student knowledge of geriatric oral care. A convenience sampling procedure was used. Sample size calculation revealed that fifty-six subjects were required to meet clinical and statistical criteria. Paired t-test addressed our hypothesis that use of the educational tool is associated with improvement in knowledge. Fifty-eight first-year dental students and nine third-year medical students completed the pre-intervention test and were given the CD-based educational tool. After seven days, all participants completed the post-intervention test. Knowledge of geriatric oral health improved among the sixty-seven students included in this study (p=0.019). When stratified on the basis of viewing the CD-ROM, the subgroup of thirty-eight students who reported not actually reviewing the CD-ROM had no change in their knowledge scores, while the subgroup of twenty-nine students who reported reviewing the CD had a significant improvement in test scores (p<0.001). Use of a self-instructional e-learning tool in geriatric oral health is effective among those students who choose to employ such tools.

Preparation and Characterization of PETI-330/Multiwalled Carbon Nanotube Composites

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Working, Dennis C.; Delozier, Donavon M.; Criss, Jim M.; Siochi, Emilie J.; Connell, John W.

2005-01-01

As part of an ongoing effort to incorporate multi-functionality into advanced composites, blends of PETI-330 and multi-walled carbon nanotubes (MWCNTs) were prepared, characterized and fabricated into moldings. The PETI-330/MWCNT mixtures were prepared at concentrations ranging from 3 to 25 weight percent by dry mixing the components in a ball mill. The resulting powders were characterized for degree of mixing, thermal and rheological properties. Based on the characterization results, PETI-330/MWCNT samples were scaled up to approx. 300 g and used to fabricate moldings 10.2 cm x 15.2 cm x 0.32 cm thick. The moldings were fabricated by injecting the mixtures at 260-280 C into a stainless steel tool followed by curing for 1 h at 371 C. The tool was designed to impart high shear during the injection process in an attempt to achieve some alignment of the MWCNTs in the flow direction. Good quality moldings were obtained that were subsequently characterized for thermal, mechanical and electrical properties. The degree of dispersion and alignment of the MWCNTs were investigated using high-resolution scanning electron microscopy and Raman spectroscopy. The preparation and preliminary characterization of PETI-330/MWCNT composites will be discussed. Keywords: phenylethynyl terminated imides, high temperature polymers, nanocomposites,

NMR methods for metabolomics of mammalian cell culture bioreactors.

PubMed

Aranibar, Nelly; Reily, Michael D

2014-01-01

Metabolomics has become an important tool for measuring pools of small molecules in mammalian cell cultures expressing therapeutic proteins. NMR spectroscopy has played an important role, largely because it requires minimal sample preparation, does not require chromatographic separation, and is quantitative. The concentrations of large numbers of small molecules in the extracellular media or within the cells themselves can be measured directly on the culture supernatant and on the supernatant of the lysed cells, respectively, and correlated with endpoints such as titer, cell viability, or glycosylation patterns. The observed changes can be used to generate hypotheses by which these parameters can be optimized. This chapter focuses on the sample preparation, data acquisition, and analysis to get the most out of NMR metabolomics data from CHO cell cultures but could easily be extended to other in vitro culture systems.

Magic Angle Spinning NMR Metabolomics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhi Hu, Jian

Nuclear Magnetic Resonance (NMR) spectroscopy is a non-destructive, quantitative, reproducible, untargeted and unbiased method that requires no or minimal sample preparation, and is one of the leading analytical tools for metabonomics research [1-3]. The easy quantification and the no need of prior knowledge about compounds present in a sample associated with NMR are advantageous over other techniques [1,4]. 1H NMR is especially attractive because protons are present in virtually all metabolites and its NMR sensitivity is high, enabling the simultaneous identification and monitoring of a wide range of low molecular weight metabolites.

Challenges of biological sample preparation for SIMS imaging of elements and molecules at subcellular resolution

NASA Astrophysics Data System (ADS)

Chandra, Subhash

2008-12-01

Secondary ion mass spectrometry (SIMS) based imaging techniques capable of subcellular resolution characterization of elements and molecules are becoming valuable tools in many areas of biology and medicine. Due to high vacuum requirements of SIMS, the live cells cannot be analyzed directly in the instrument. The sample preparation, therefore, plays a critical role in preserving the native chemical composition for SIMS analysis. This work focuses on the evaluation of frozen-hydrated and frozen freeze-dried sample preparations for SIMS studies of cultured cells with a CAMECA IMS-3f dynamic SIMS ion microscope instrument capable of producing SIMS images with a spatial resolution of 500 nm. The sandwich freeze-fracture method was used for fracturing the cells. The complimentary fracture planes in the plasma membrane were characterized by field-emission secondary electron microscopy (FESEM) in the frozen-hydrated state. The cells fractured at the dorsal surface were used for SIMS analysis. The frozen-hydrated SIMS analysis of individual cells under dynamic primary ion beam (O 2+) revealed local secondary ion signal enhancements correlated with the water image signals of 19(H 3O) +. A preferential removal of water from the frozen cell matrix in the Z-axis was also observed. These complications render the frozen-hydrated sample type less desirable for subcellular dynamic SIMS studies. The freeze-drying of frozen-hydrated cells, either inside the instrument or externally in a freeze-drier, allowed SIMS imaging of subcellular chemical composition. Morphological evaluations of fractured freeze-dried cells with SEM and confocal laser scanning microscopy (CLSM) revealed well-preserved mitochondria, Golgi apparatus, and stress fibers. SIMS analysis of fractured freeze-dried cells revealed well-preserved chemical composition of even the most highly diffusible ions like K + and Na + in physiologically relevant concentrations. The high K-low Na signature in individual cells provided a rule-of-thumb criterion for the validation of sample preparation. The fractured freeze-dried cells allowed 3-D SIMS imaging and localization of 13C 15N labeled molecules and therapeutic drugs containing an elemental tag. Examples are shown to demonstrate that both diffusible elements and molecules are prone to artifact-induced relocation at subcellular scale if the sample preparation is compromised. The sample preparation is problem dependent and may vary widely between the diverse sample types of biological systems and the type of instrument used for SIMS analysis. The sample preparation, however, must be validated so that SIMS can be applied with confidence in biology and medicine.

40 CFR 51.5 - What tools are available to help prepare and report emissions data?

Code of Federal Regulations, 2014 CFR

2014-07-01

... PLANS Air Emissions Reporting Requirements General Information for Inventory Preparers § 51.5 What tools... 40 Protection of Environment 2 2014-07-01 2014-07-01 false What tools are available to help prepare and report emissions data? 51.5 Section 51.5 Protection of Environment ENVIRONMENTAL PROTECTION...

40 CFR 51.5 - What tools are available to help prepare and report emissions data?

Code of Federal Regulations, 2012 CFR

2012-07-01

... PLANS Air Emissions Reporting Requirements General Information for Inventory Preparers § 51.5 What tools... 40 Protection of Environment 2 2012-07-01 2012-07-01 false What tools are available to help prepare and report emissions data? 51.5 Section 51.5 Protection of Environment ENVIRONMENTAL PROTECTION...

40 CFR 51.5 - What tools are available to help prepare and report emissions data?

Code of Federal Regulations, 2013 CFR

2013-07-01

... PLANS Air Emissions Reporting Requirements General Information for Inventory Preparers § 51.5 What tools... 40 Protection of Environment 2 2013-07-01 2013-07-01 false What tools are available to help prepare and report emissions data? 51.5 Section 51.5 Protection of Environment ENVIRONMENTAL PROTECTION...

The diameter of nanotubes formed on Ti-6Al-4V alloy controls the adhesion and differentiation of Saos-2 cells

PubMed Central

Filova, Elena; Fojt, Jaroslav; Kryslova, Marketa; Moravec, Hynek; Joska, Ludek; Bacakova, Lucie

2015-01-01

Ti-6Al-4V-based nanotubes were prepared on a Ti-6Al-4V surface by anodic oxidation on 10 V, 20 V, and 30 V samples. The 10 V, 20 V, and 30 V samples and a control smooth Ti-6Al-4V sample were evaluated in terms of their chemical composition, diameter distribution, and cellular response. The surfaces of the 10 V, 20 V, and 30 V samples consisted of nanotubes of a relatively wide range of diameters that increased with the voltage. Saos-2 cells had a similar initial adhesion on all nanotube samples to the control Ti-6Al-4V sample, but it was lower than on glass. On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes. On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples. The final cellular densities on 10 V, 20 V, and 30 V samples were higher than on glass. Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties. PMID:26648719

An efficient and sensitive method for preparing cDNA libraries from scarce biological samples

PubMed Central

Sterling, Catherine H.; Veksler-Lublinsky, Isana; Ambros, Victor

2015-01-01

The preparation and high-throughput sequencing of cDNA libraries from samples of small RNA is a powerful tool to quantify known small RNAs (such as microRNAs) and to discover novel RNA species. Interest in identifying the small RNA repertoire present in tissues and in biofluids has grown substantially with the findings that small RNAs can serve as indicators of biological conditions and disease states. Here we describe a novel and straightforward method to clone cDNA libraries from small quantities of input RNA. This method permits the generation of cDNA libraries from sub-picogram quantities of RNA robustly, efficiently and reproducibly. We demonstrate that the method provides a significant improvement in sensitivity compared to previous cloning methods while maintaining reproducible identification of diverse small RNA species. This method should have widespread applications in a variety of contexts, including biomarker discovery from scarce samples of human tissue or body fluids. PMID:25056322

Acrylamide content in French fries prepared in households: A pilot study in Spanish homes.

PubMed

Mesias, Marta; Delgado-Andrade, Cristina; Holgado, Francisca; Morales, Francisco J

2018-09-15

An observational cross-sectional pilot study in 73 Spanish households was conducted to evaluate the impact of consumer practices on the formation of acrylamide during the preparation of French fries from fresh potatoes applying one stage frying. 45.2% of samples presented acrylamide concentrations above the benchmark level for French fries (500 µg/kg). 6.9% of samples exceeded 2000 µg/kg and the 95th percentile was 2028 µg/kg. The median and average values were significantly higher than the EFSA report for this food category, suggesting that the total exposure to acrylamide by the population could be underestimated. In this randomised scenario of cooking practices, the content of reducing sugar and asparagine did not explain the acrylamide levels. However, the chromatic parameter a ∗ of the fried potato was a powerful tool to classify the samples according to the acrylamide benchmark level regardless of the agronomical characteristics of the potato or the consumer practices. Copyright © 2018 Elsevier Ltd. All rights reserved.

Advances in ultrasensitive mass spectrometry of organic molecules.

PubMed

Kandiah, Mathivathani; Urban, Pawel L

2013-06-21

Ultrasensitive mass spectrometric analysis of organic molecules is important for various branches of chemistry, and other fields including physics, earth and environmental sciences, archaeology, biomedicine, and materials science. It finds applications--as an enabling tool--in systems biology, biological imaging, clinical analysis, and forensics. Although there are a number of technical obstacles associated with the analysis of samples by mass spectrometry at ultratrace level (for example analyte losses during sample preparation, insufficient sensitivity, ion suppression), several noteworthy developments have been made over the years. They include: sensitive ion sources, loss-free interfaces, ion optics components, efficient mass analyzers and detectors, as well as "smart" sample preparation strategies. Some of the mass spectrometric methods published to date can achieve sensitivity which is by several orders of magnitude higher than that of alternative approaches. Femto- and attomole level limits of detection are nowadays common, while zepto- and yoctomole level limits of detection have also been reported. We envision that the ultrasensitive mass spectrometric assays will soon contribute to new discoveries in bioscience and other areas.

Ultrasound: a subexploited tool for sample preparation in metabolomics.

PubMed

Luque de Castro, M D; Delgado-Povedano, M M

2014-01-02

Metabolomics, one of the most recently emerged "omics", has taken advantage of ultrasound (US) to improve sample preparation (SP) steps. The metabolomics-US assisted SP step binomial has experienced a dissimilar development that has depended on the area (vegetal or animal) and the SP step. Thus, vegetal metabolomics and US assisted leaching has received the greater attention (encompassing subdisciplines such as metallomics, xenometabolomics and, mainly, lipidomics), but also liquid-liquid extraction and (bio)chemical reactions in metabolomics have taken advantage of US energy. Also clinical and animal samples have benefited from US assisted SP in metabolomics studies but in a lesser extension. The main effects of US have been shortening of the time required for the given step, and/or increase of its efficiency or availability for automation; nevertheless, attention paid to potential degradation caused by US has been scant or nil. Achievements and weak points of the metabolomics-US assisted SP step binomial are discussed and possible solutions to the present shortcomings are exposed. Copyright © 2013 Elsevier B.V. All rights reserved.

Fingerprinting and characterization of anthocyanins in 94 colored wheat varieties and blue aleurone and purple pericarp wheat crosses.

PubMed

Krüger, Stephanie; Morlock, Gertrud E

2018-02-23

Colored wheat varieties and crosses were analyzed to figure out their anthocyanin profiles, and thus, their potential as health-related food. After method development, the obtained 94 anthocyanin fingerprints allowed the clear differentiation of the blue aleurone and purple pericarp genotypes as well as their breeding lines. The method was trimmed so that the complete analysis of the whole grain flour including sample preparation of up to 20 samples on one plate took less than 3 h (<9 min per sample) and total costs including sample preparation were <1.0 Euro/sample. Sample preparation of the complex wheat matrix was reduced to a minimum (only acidified methanol extraction of the ground whole wheat grain). Separation was well achieved on amino phases with a mixture of ethyl acetate, 2-butanone, water and formic acid. It was superior to the separation on either normal or reversed phases and more robust with regard to intrinsic pH variances of the sample extracts. Pattern recognition of anthocyanins was simply performed by visual detection (the image), a key feature of high-performance thin-layer chromatography. Wheat varieties and crosses with higher anthocyanin contents were easily selectable, and thus, successfully made out. Prominent anthocyanin zones were characterized by electrospray ionization mass spectrometry. Their sugar moiety was characterized via methanolysis and compared with the sugars available freely in the whole wheat grain. The developed profiling is a fast and efficient screening tool with option for quantification or identification on the same HPTLC plate. Copyright © 2018 Elsevier B.V. All rights reserved.

Experiences of using the GMP audit preparation tool in pharmaceutical contract manufacturer audits.

PubMed

Linna, Anu; Korhonen, Mirka; Airaksinen, Marja; Juppo, Anne Mari

2010-06-01

Use of external contractors is nowadays inevitable in the pharmaceutical industry. Therefore the amount of current good manufacturing practice audits has been increasing. During the audit, a large amount of items should be covered in a limited amount of time. Consequently, pharmaceutical companies should have systematic and effective ways to manage and prepare for the audits. This study is a continuation to the earlier study, where a tool for the preparation of cGMP audit was developed and its content was validated. The objective of this study was to evaluate the usefulness of the developed tool in audit preparation and during the actual cGMP audit. Three qualitative research methods were used in this study (observation, interviews, and opinion survey). First, the validity of the information given through the tool was examined by comparing the responses to the actual conditions observed during the contract manufacturer audits (n = 15). Additionally the opinions of the contract manufacturers of the tool were gathered (n = 10) and the auditors were interviewed (n = 2). The developed tool was proven to be useful in audit preparation phase from both the auditor's and the contract manufacturers' point of view. Furthermore, using the tool can also save some time when performing the audit. The results show that using the tool can give significant support in audit preparation phase and also during the actual audit.

Foodomics and Food Safety: Where We Are.

PubMed

Andjelković, Uroš; Šrajer Gajdošik, Martina; Gašo-Sokač, Dajana; Martinović, Tamara; Josić, Djuro

2017-09-01

The power of foodomics as a discipline that is now broadly used for quality assurance of food products and adulteration identification, as well as for determining the safety of food, is presented. Concerning sample preparation and application, maintenance of highly sophisticated instruments for both high-performance and high-throughput techniques, and analysis and data interpretation, special attention has to be paid to the development of skilled analysts. The obtained data shall be integrated under a strong bioinformatics environment. Modern mass spectrometry is an extremely powerful analytical tool since it can provide direct qualitative and quantitative information about a molecule of interest from only a minute amount of sample. Quality of this information is influenced by the sample preparation procedure, the type of mass spectrometer used and the analyst's skills. Technical advances are bringing new instruments of increased sensitivity, resolution and speed to the market. Other methods presented here give additional information and can be used as complementary tools to mass spectrometry or for validation of obtained results. Genomics and transcriptomics, as well as affinity-based methods, still have a broad use in food analysis. Serious drawbacks of some of them, especially the affinity-based methods, are the cross-reactivity between similar molecules and the influence of complex food matrices. However, these techniques can be used for pre-screening in order to reduce the large number of samples. Great progress has been made in the application of bioinformatics in foodomics. These developments enabled processing of large amounts of generated data for both identification and quantification, and for corresponding modeling.

Foodomics and Food Safety: Where We Are

PubMed Central

Andjelković, Uroš

2017-01-01

Summary The power of foodomics as a discipline that is now broadly used for quality assurance of food products and adulteration identification, as well as for determining the safety of food, is presented. Concerning sample preparation and application, maintenance of highly sophisticated instruments for both high-performance and high-throughput techniques, and analysis and data interpretation, special attention has to be paid to the development of skilled analysts. The obtained data shall be integrated under a strong bioinformatics environment. Modern mass spectrometry is an extremely powerful analytical tool since it can provide direct qualitative and quantitative information about a molecule of interest from only a minute amount of sample. Quality of this information is influenced by the sample preparation procedure, the type of mass spectrometer used and the analyst’s skills. Technical advances are bringing new instruments of increased sensitivity, resolution and speed to the market. Other methods presented here give additional information and can be used as complementary tools to mass spectrometry or for validation of obtained results. Genomics and transcriptomics, as well as affinity-based methods, still have a broad use in food analysis. Serious drawbacks of some of them, especially the affinity-based methods, are the cross-reactivity between similar molecules and the influence of complex food matrices. However, these techniques can be used for pre-screening in order to reduce the large number of samples. Great progress has been made in the application of bioinformatics in foodomics. These developments enabled processing of large amounts of generated data for both identification and quantification, and for corresponding modeling. PMID:29089845

A Multicenter Study To Evaluate the Performance of High-Throughput Sequencing for Virus Detection

PubMed Central

Ng, Siemon H. S.; Vandeputte, Olivier; Aljanahi, Aisha; Deyati, Avisek; Cassart, Jean-Pol; Charlebois, Robert L.; Taliaferro, Lanyn P.

2017-01-01

ABSTRACT The capability of high-throughput sequencing (HTS) for detection of known and unknown viruses makes it a powerful tool for broad microbial investigations, such as evaluation of novel cell substrates that may be used for the development of new biological products. However, like any new assay, regulatory applications of HTS need method standardization. Therefore, our three laboratories initiated a study to evaluate performance of HTS for potential detection of viral adventitious agents by spiking model viruses in different cellular matrices to mimic putative materials for manufacturing of biologics. Four model viruses were selected based upon different physical and biochemical properties and commercial availability: human respiratory syncytial virus (RSV), Epstein-Barr virus (EBV), feline leukemia virus (FeLV), and human reovirus (REO). Additionally, porcine circovirus (PCV) was tested by one laboratory. Independent samples were prepared for HTS by spiking intact viruses or extracted viral nucleic acids, singly or mixed, into different HeLa cell matrices (resuspended whole cells, cell lysate, or total cellular RNA). Data were obtained using different sequencing platforms (Roche 454, Illumina HiSeq1500 or HiSeq2500). Bioinformatic analyses were performed independently by each laboratory using available tools, pipelines, and databases. The results showed that comparable virus detection was obtained in the three laboratories regardless of sample processing, library preparation, sequencing platform, and bioinformatic analysis: between 0.1 and 3 viral genome copies per cell were detected for all of the model viruses used. This study highlights the potential for using HTS for sensitive detection of adventitious viruses in complex biological samples containing cellular background. IMPORTANCE Recent high-throughput sequencing (HTS) investigations have resulted in unexpected discoveries of known and novel viruses in a variety of sample types, including research materials, clinical materials, and biological products. Therefore, HTS can be a powerful tool for supplementing current methods for demonstrating the absence of adventitious or unwanted viruses in biological products, particularly when using a new cell line. However, HTS is a complex technology with different platforms, which needs standardization for evaluation of biologics. This collaborative study was undertaken to investigate detection of different virus types using two different HTS platforms. The results of the independently performed studies demonstrated a similar sensitivity of virus detection, regardless of the different sample preparation and processing procedures and bioinformatic analyses done in the three laboratories. Comparable HTS detection of different virus types supports future development of reference virus materials for standardization and validation of different HTS platforms. PMID:28932815

A Critical Review on Clinical Application of Separation Techniques for Selective Recognition of Uracil and 5-Fluorouracil.

PubMed

Pandey, Khushaboo; Dubey, Rama Shankar; Prasad, Bhim Bali

2016-03-01

The most important objectives that are frequently found in bio-analytical chemistry involve applying tools to relevant medical/biological problems and refining these applications. Developing a reliable sample preparation step, for the medical and biological fields is another primary objective in analytical chemistry, in order to extract and isolate the analytes of interest from complex biological matrices. Since, main inborn errors of metabolism (IEM) diagnosable through uracil analysis and the therapeutic monitoring of toxic 5-fluoruracil (an important anti-cancerous drug) in dihydropyrimidine dehydrogenase deficient patients, require an ultra-sensitive, reproducible, selective, and accurate analytical techniques for their measurements. Therefore, keeping in view, the diagnostic value of uracil and 5-fluoruracil measurements, this article refines several analytical techniques involved in selective recognition and quantification of uracil and 5-fluoruracil from biological and pharmaceutical samples. The prospective study revealed that implementation of molecularly imprinted polymer as a solid-phase material for sample preparation and preconcentration of uracil and 5-fluoruracil had proven to be effective as it could obviates problems related to tedious separation techniques, owing to protein binding and drastic interferences, from the complex matrices in real samples such as blood plasma, serum samples.

Validation of a score tool for measurement of histological severity in juvenile dermatomyositis and association with clinical severity of disease

PubMed Central

Varsani, Hemlata; Charman, Susan C; Li, Charles K; Marie, Suely K N; Amato, Anthony A; Banwell, Brenda; Bove, Kevin E; Corse, Andrea M; Emslie-Smith, Alison M; Jacques, Thomas S; Lundberg, Ingrid E; Minetti, Carlo; Nennesmo, Inger; Rushing, Elisabeth J; Sallum, Adriana M E; Sewry, Caroline; Pilkington, Clarissa A; Holton, Janice L; Wedderburn, Lucy R

2015-01-01

Objectives To study muscle biopsy tissue from patients with juvenile dermatomyositis (JDM) in order to test the reliability of a score tool designed to quantify the severity of histological abnormalities when applied to biceps humeri in addition to quadriceps femoris. Additionally, to evaluate whether elements of the tool correlate with clinical measures of disease severity. Methods 55 patients with JDM with muscle biopsy tissue and clinical data available were included. Biopsy samples (33 quadriceps, 22 biceps) were prepared and stained using standardised protocols. A Latin square design was used by the International Juvenile Dermatomyositis Biopsy Consensus Group to score cases using our previously published score tool. Reliability was assessed by intraclass correlation coefficient (ICC) and scorer agreement (α) by assessing variation in scorers’ ratings. Scores from the most reliable tool items correlated with clinical measures of disease activity at the time of biopsy. Results Inter- and intraobserver agreement was good or high for many tool items, including overall assessment of severity using a Visual Analogue Scale. The tool functioned equally well on biceps and quadriceps samples. A modified tool using the most reliable score items showed good correlation with measures of disease activity. Conclusions The JDM biopsy score tool has high inter- and intraobserver agreement and can be used on both biceps and quadriceps muscle tissue. Importantly, the modified tool correlates well with clinical measures of disease activity. We propose that standardised assessment of muscle biopsy tissue should be considered in diagnostic investigation and clinical trials in JDM. PMID:24064003

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement.

PubMed

Liu, Jun; Wilson, John; Davis, Claire; Peyton, Anthony

2017-11-07

This paper elaborates the sample preparation protocols required to obtain optimal domain patterns using the Bitter method, focusing on the extra steps compared to standard metallographic sample preparation procedures. The paper proposes a novel bespoke rig for dynamic domain imaging with in situ BH (magnetic hysteresis) measurements and elaborates the protocols for the sensor preparation and the use of the rig to ensure accurate BH measurement. The protocols for static and ordinary dynamic domain imaging (without in situ BH measurements) are also presented. The reported method takes advantage of the convenience and high sensitivity of the traditional Bitter method and enables in situ BH measurement without interrupting or interfering with the domain wall movement processes. This facilitates establishing a direct and quantitative link between the domain wall movement processes-microstructural feature interactions in ferritic steels with their BH loops. This method is anticipated to become a useful tool for the fundamental study of microstructure-magnetic property relationships in steels and to help interpret the electromagnetic sensor signals for non-destructive evaluation of steel microstructures.

Flat ion milling: a powerful tool for preparation of cross-sections of lead-silver alloys.

PubMed

Brodusch, Nicolas; Boisvert, Sophie; Gauvin, Raynald

2013-06-01

While conventional mechanical and chemical polishing results in stress, deformation and polishing particles embedded on the surface, flat milling with Ar+ ions erodes the material with no mechanical artefacts. This flat milling process is presented as an alternative method to prepare a Pb-Ag alloy cross-section for scanning electron microscopy. The resulting surface is free of scratches with very little to no stress induced, so that electron diffraction and channelling contrast are possible. The results have shown that energy dispersive spectrometer (EDS) mapping, electron channelling contrast imaging and electron backscatter diffraction can be conducted with only one sample preparation step. Electron diffraction patterns acquired at 5 keV possessed very good pattern quality, highlighting an excellent surface condition. An orientation map was acquired at 20 keV with an indexing rate of 90.1%. An EDS map was performed at 5 keV, and Pb-Ag precipitates of sizes lower than 100 nm were observed. However, the drawback of the method is the generation of a noticeable surface topography resulting from the interaction of the ion beam with a polycrystalline and biphasic sample.

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement

PubMed Central

Liu, Jun; Wilson, John; Davis, Claire; Peyton, Anthony

2017-01-01

This paper elaborates the sample preparation protocols required to obtain optimal domain patterns using the Bitter method, focusing on the extra steps compared to standard metallographic sample preparation procedures. The paper proposes a novel bespoke rig for dynamic domain imaging with in situ BH (magnetic hysteresis) measurements and elaborates the protocols for the sensor preparation and the use of the rig to ensure accurate BH measurement. The protocols for static and ordinary dynamic domain imaging (without in situ BH measurements) are also presented. The reported method takes advantage of the convenience and high sensitivity of the traditional Bitter method and enables in situ BH measurement without interrupting or interfering with the domain wall movement processes. This facilitates establishing a direct and quantitative link between the domain wall movement processes–microstructural feature interactions in ferritic steels with their BH loops. This method is anticipated to become a useful tool for the fundamental study of microstructure–magnetic property relationships in steels and to help interpret the electromagnetic sensor signals for non-destructive evaluation of steel microstructures. PMID:29155796

Analyzing Immunoglobulin Repertoires

PubMed Central

Chaudhary, Neha; Wesemann, Duane R.

2018-01-01

Somatic assembly of T cell receptor and B cell receptor (BCR) genes produces a vast diversity of lymphocyte antigen recognition capacity. The advent of efficient high-throughput sequencing of lymphocyte antigen receptor genes has recently generated unprecedented opportunities for exploration of adaptive immune responses. With these opportunities have come significant challenges in understanding the analysis techniques that most accurately reflect underlying biological phenomena. In this regard, sample preparation and sequence analysis techniques, which have largely been borrowed and adapted from other fields, continue to evolve. Here, we review current methods and challenges of library preparation, sequencing and statistical analysis of lymphocyte receptor repertoire studies. We discuss the general steps in the process of immune repertoire generation including sample preparation, platforms available for sequencing, processing of sequencing data, measurable features of the immune repertoire, and the statistical tools that can be used for analysis and interpretation of the data. Because BCR analysis harbors additional complexities, such as immunoglobulin (Ig) (i.e., antibody) gene somatic hypermutation and class switch recombination, the emphasis of this review is on Ig/BCR sequence analysis. PMID:29593723

Small-scale enzymatic digestion of glycoproteins and proteoglycans for analysis of oligosaccharides by LC-MS and FACE gel electrophoresis.

PubMed

Estrella, Ruby P; Whitelock, John M; Roubin, Rebecca H; Packer, Nicolle H; Karlsson, Niclas G

2009-01-01

Structural characterization of oligosaccharides from proteoglycans and other glycoproteins is greatly enhanced through the use of mass spectrometry and gel electrophoresis. Sample preparation for these sensitive techniques often requires enzymatic treatments to produce oligosaccharide sequences for subsequent analysis. This chapter describes several small-scale methods for in-gel, on-blot, and in-solution enzymatic digestions in preparation for graphitized carbon liquid chromatography-mass spectrometry (LC-MS) analysis, with specific applications indicated for glycosaminoglycans (GAGs) and N-linked oligosaccharides. In addition, accompanying procedures for oligosaccharide reduction by sodium borohydride, sample desalting via carbon microcolumn, desialylation by sialidase enzyme treatment, and small-scale oligosaccharide species fractionation are included. Fluorophore-assisted carbohydrate electrophoresis (FACE) is another useful method to isolate derivatized oligosaccharides. Overall, the modularity of these techniques provides ease and flexibility for use in conjunction with mass spectrometric and electrophoretic tools for glycomic research studies.

Crystallization of PTP Domains.

PubMed

Levy, Colin; Adams, James; Tabernero, Lydia

2016-01-01

Protein crystallography is the most powerful method to obtain atomic resolution information on the three-dimensional structure of proteins. An essential step towards determining the crystallographic structure of a protein is to produce good quality crystals from a concentrated sample of purified protein. These crystals are then used to obtain X-ray diffraction data necessary to determine the 3D structure by direct phasing or molecular replacement if the model of a homologous protein is available. Here, we describe the main approaches and techniques to obtain suitable crystals for X-ray diffraction. We include tools and guidance on how to evaluate and design the protein construct, how to prepare Se-methionine derivatized protein, how to assess the stability and quality of the sample, and how to crystallize and prepare crystals for diffraction experiments. While general strategies for protein crystallization are summarized, specific examples of the application of these strategies to the crystallization of PTP domains are discussed.

Recent development in software and automation tools for high-throughput discovery bioanalysis.

PubMed

Shou, Wilson Z; Zhang, Jun

2012-05-01

Bioanalysis with LC-MS/MS has been established as the method of choice for quantitative determination of drug candidates in biological matrices in drug discovery and development. The LC-MS/MS bioanalytical support for drug discovery, especially for early discovery, often requires high-throughput (HT) analysis of large numbers of samples (hundreds to thousands per day) generated from many structurally diverse compounds (tens to hundreds per day) with a very quick turnaround time, in order to provide important activity and liability data to move discovery projects forward. Another important consideration for discovery bioanalysis is its fit-for-purpose quality requirement depending on the particular experiments being conducted at this stage, and it is usually not as stringent as those required in bioanalysis supporting drug development. These aforementioned attributes of HT discovery bioanalysis made it an ideal candidate for using software and automation tools to eliminate manual steps, remove bottlenecks, improve efficiency and reduce turnaround time while maintaining adequate quality. In this article we will review various recent developments that facilitate automation of individual bioanalytical procedures, such as sample preparation, MS/MS method development, sample analysis and data review, as well as fully integrated software tools that manage the entire bioanalytical workflow in HT discovery bioanalysis. In addition, software tools supporting the emerging high-resolution accurate MS bioanalytical approach are also discussed.

DFAS Needs More Effective Controls Over Managing DoD Contractor Debt

DTIC Science & Technology

2011-07-15

Acrobat fil e onl y) to audc lev@dod ig.m il. Copies of management comments must have the actual signature of the authori zing official for yo ur...Attribute Testing Results 18 E. Sampling Methodology 20 Glossary of Technical Terms 21 Management...Bill of Collection number to each contractor debt as a debt management tool. CDS allows users to prepare a demand letter from the database, post

Understanding the Potential Content and Structure of an International Convention on the Human Rights of People with Disabilities: Sample Treaty Provisions Drawn from Existing International Instruments. A Reference Tool.

ERIC Educational Resources Information Center

Lord, Janet E.

This document is designed to prepare advocates in the international disability community for productive participation in the development of international conventions on the human rights of people with disabilities. Knowledge of the standard categories of international law provisions will help participants address issues related to the structure of…

Fourier Transform Infrared Spectroscopy as a Tool in Analysis of Proteus mirabilis Endotoxins.

PubMed

Żarnowiec, Paulina; Czerwonka, Grzegorz; Kaca, Wiesław

2017-01-01

Fourier transform infrared spectroscopy (FT-IR) was used to scan whole bacterial cells as well as lipopolysaccharides (LPSs, endotoxins) isolated from them. Proteus mirabilis cells, with chemically defined LPSs, served as a model for the ATR FT-IR method. The paper focuses on three steps of infrared spectroscopy: (1) sample preparation, (2) IR scanning, and (3) multivariate analysis of IR data (principal component analysis, PCA).

Detection of caffeine in tea, instant coffee, green tea beverage, and soft drink by direct analysis in real time (DART) source coupled to single-quadrupole mass spectrometry.

PubMed

Wang, Lei; Zhao, Pengyue; Zhang, Fengzu; Bai, Aijuan; Pan, Canping

2013-01-01

Ambient ionization direct analysis in real time (DART) coupled to single-quadrupole MS (DART-MS) was evaluated for rapid detection of caffeine in commercial samples without chromatographic separation or sample preparation. Four commercial samples were examined: tea, instant coffee, green tea beverage, and soft drink. The response-related parameters were optimized for the DART temperature and MS fragmentor. Under optimal conditions, the molecular ion (M+H)+ was the major ion for identification of caffeine. The results showed that DART-MS is a promising tool for the quick analysis of important marker molecules in commercial samples. Furthermore, this system has demonstrated significant potential for high sample throughput and real-time analysis.

Integration of Gas Chromatography Mass Spectrometry Methods for Differentiating Ricin Preparation Methods

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wunschel, David S.; Melville, Angela M.; Ehrhardt, Christopher J.

2012-05-17

The investigation of crimes involving chemical or biological agents is infrequent, but presents unique analytical challenges. The protein toxin ricin is encountered more frequently than other agents and is found in the seeds of the castor plant Ricinus communis. Typically, the toxin is extracted from castor seeds utilizing a variety of different recipes that result in varying purity of the toxin. Moreover, these various purification steps can also leave or differentially remove a variety of exogenous and endogenous residual components with the toxin that may indicate the type and number of purification steps involved. We have applied three gas chromatographicmore » - mass spectrometric (GC-MS) based analytical methods to measure the variation in seed carbohydrates and castor oil ricinoleic acid as well as the presence of solvents used for purification. These methods were applied to the same samples prepared using four previously identified toxin preparation methods starting from four varieties of castor seeds. The individual data sets for seed carbohydrate profiles, ricinoleic acid or acetone amount each provided information capable of differentiating different types of toxin preparations across seed types. However, the integration of the data sets using multivariate factor analysis provided a clear distinction of all samples based on the preparation method and independent of the seed source. In particular the abundance of mannose, arabinose, fucose, ricinoleic acid and acetone were shown to be important differentiating factors. These complementary tools provide a more confident determination of the method of toxin preparation.« less

Microstructural Study of Micron-Sized Craters Simulating Stardust Impacts in Aluminum 1100 Targets

NASA Technical Reports Server (NTRS)

Leroux, Hugues; Borg, Janet; Troadec, David; Djouadi, Zahia; Horz, Friedrich

2006-01-01

Various microscopic techniques were used to characterize experimental micro- craters in aluminium foils to prepare for the comprehensive analysis of the cometary and interstellar particle impacts in aluminium foils to be returned by the Stardust mission. First, SEM (Scanning Electron Microscopy) and EDS (Energy Dispersive X-ray Spectroscopy) were used to study the morphology of the impact craters and the bulk composition of the residues left by soda-lime glass impactors. A more detailed structural and compositional study of impactor remnants was then performed using TEM (Transmission Electron Microscopy), EDS, and electron diffraction methods. The TEM samples were prepared by Focused Ion Beam (FIB) methods. This technique proved to be especially valuable in studying impact crater residues and impact crater morphology. Finally, we also showed that InfraRed microscopy (IR) can be a quick and reliable tool for such investigations. The combination of all of these tools enables a complete microscopic characterization of the craters.

Evaluating the impact of a pre-rotation workshop on student preparation for clinical advanced pharmacy practice experiences.

PubMed

Medina, Melissa S; Stark, Jennifer E; Vesta, Kimi S; Lockhart, Staci M

2008-10-01

This pilot study was designed to evaluate the impact of a pre-rotation workshop (PRW) on pharmacy students' clinical skills and preparation for clinical Advanced Pharmacy Practice Experiences (APPE) involving direct patient care. Randomized controlled trial of an educational intervention with Institutional Review Board approval. PRW activities designed to simulate rotation activities around five competencies, patient charts, medication histories, SOAP notes, patient presentations, and professionalism. Endpoints were evaluated using clinical rotation preceptors' evaluation of performance and students' performance on objective structured clinical exams (OSCE). Eight fourth-year students and eight GPA matched controls (20% of the total class) were selected to voluntarily participate. The PRW demonstrated a positive impact on students' clinical skills and preparation for rotations by improving OSCE performance. However, no significant differences were found between groups when comparing preceptor evaluations of skills on rotations. These results are limited by the small sample size, potential OSCE "test-wiseness" effects, lack of OSCE evaluator blinding to study groups, potential case specificity effects due to the limited number of cases used on the OSCE and possible lack of sensitivity of the rotation evaluation tool to capture true differences among the experimental and control group participants. The PRW was successful at advancing students' clinical skills and preparation for rotations and may be considered as a tool to help bridge didactic to clinical experiences in the Pharm.D. curriculum.

Online friends, offline loved ones, and full-time media: young adult "mass personal" use of communication resources for informational and emotional support.

PubMed

Love, Brad; Donovan, Erin E

2014-06-01

As Web 2.0 technologies proliferate, patient education is changing dramatically. Information about prevention and survivorship arrives from a mix of sources. The present manuscript describes a study to shed light on how young adults (YAs) affected by cancer manage the digital world. Our investigation was guided by a research question asking how young adults affected by cancer engage in communication work in an environment of mass personal communication. The sample for this research consisted of 500 posts comprising 50 complete threads from an online support community for young adults affected by cancer. Threads were purposively sampled in a multi-stage process. Researchers used constant comparison to define themes, examining text in increments. Individuals harnessed assets of various communication tools for the purposes of message preparation and credibility checking. YAs demonstrated the multi-channel way they move between channels for different purposes, driven by preparation for future interactions. The result is a process that allows co-creation of knowledge in a trusted community. Findings indicate that completing communication work through multiple channels in a deliberate and savvy way is normal for YAs, particularly for message preparation and credibility checking. The multidirectional nature of digital tools plays an important role for YAs, as interactive resources appear to be the first or second stop for information after key events in the cancer trajectory. Results from this study are important as guidance to help manage the volume and depth of information common to the cancer experience in the Web 2.0 world.

Optimizing Frozen Sample Preparation for Laser Microdissection: Assessment of CryoJane Tape-Transfer System®

PubMed Central

Golubeva, Yelena G.; Smith, Roberta M.; Sternberg, Lawrence R.

2013-01-01

Laser microdissection is an invaluable tool in medical research that facilitates collecting specific cell populations for molecular analysis. Diversity of research targets (e.g., cancerous and precancerous lesions in clinical and animal research, cell pellets, rodent embryos, etc.) and varied scientific objectives, however, present challenges toward establishing standard laser microdissection protocols. Sample preparation is crucial for quality RNA, DNA and protein retrieval, where it often determines the feasibility of a laser microdissection project. The majority of microdissection studies in clinical and animal model research are conducted on frozen tissues containing native nucleic acids, unmodified by fixation. However, the variable morphological quality of frozen sections from tissues containing fat, collagen or delicate cell structures can limit or prevent successful harvest of the desired cell population via laser dissection. The CryoJane Tape-Transfer System®, a commercial device that improves cryosectioning outcomes on glass slides has been reported superior for slide preparation and isolation of high quality osteocyte RNA (frozen bone) during laser dissection. Considering the reported advantages of CryoJane for laser dissection on glass slides, we asked whether the system could also work with the plastic membrane slides used by UV laser based microdissection instruments, as these are better suited for collection of larger target areas. In an attempt to optimize laser microdissection slide preparation for tissues of different RNA stability and cryosectioning difficulty, we evaluated the CryoJane system for use with both glass (laser capture microdissection) and membrane (laser cutting microdissection) slides. We have established a sample preparation protocol for glass and membrane slides including manual coating of membrane slides with CryoJane solutions, cryosectioning, slide staining and dissection procedure, lysis and RNA extraction that facilitated efficient dissection and high quality RNA retrieval from CryoJane preparations. CryoJane technology therefore has the potential to facilitate standardization of laser microdissection slide preparation from frozen tissues. PMID:23805281

Chemometric tool for identification of iron-gall inks by use of visible-near infrared fibre optic reflection spectroscopy.

PubMed

Gál, Lukáš; Čeppan, Michal; Reháková, Milena; Dvonka, Vladimír; Tarajčáková, Jarmila; Hanus, Jozef

2013-11-01

A method has been developed for identification of corrosive iron-gall inks in historical drawings and documents. The method is based on target-factor analysis of visible-near infrared fibre optic reflection spectra (VIS-NIR FORS). A set of reference spectra was obtained from model samples of laboratory-prepared inks covering a wide range of mixing ratios of basic ink components deposited on substrates and artificially aged. As criteria for correspondence of a studied spectrum with a reference spectrum, the apparent error in target (AET) and the empirical function SPOIL according to Malinowski were used. The capability of the proposed tool to distinguish corrosive iron-gall inks from bistre and sepia inks was evaluated by use of a set of control samples of bistre, sepia, and iron-gall inks. Examples are presented of analysis of historical drawings from the 15th and 16th centuries and written documents from the 19th century. The results of analysis based on the tool were confirmed by XRF analysis and colorimetric spot analysis.

Comparing paraffined and deparaffinized breast cancer tissue samples and an analysis of Raman spectroscopy and infrared methods

NASA Astrophysics Data System (ADS)

Depciuch, J.; Kaznowska, E.; Szmuc, K.; Zawlik, I.; Cholewa, M.; Heraud, P.; Cebulski, J.

2016-05-01

Breast cancer makes up a quarter of all cancer in women, which is why research into new diagnostic methods and sample preparations need to be developed at an accelerated pace. Researchers are looking for diagnostic tools to detect when an individual has cancer cells and use that information to see what measurements and approaches can be used to take further diagnostic steps. The most common method of sample preparation is the imbibing of tumor tissue in paraffin, which can produce a background for spectroscopic measurements in the range of 500-3500 cm-1. In this study we demonstrated that proper preparation of paraffin-embedded specimens and the measurement methodology can eliminate paraffin vibration, as was done in the work Depciuch et al. 2015. Thanks to this spectroscopic technique there may become a reliable and accurate method of diagnosing breast cancer based on the evidence found from the prepared samples. The study compared the results obtained through Raman spectroscopy and FTIR (Fourier Transform Infrared) measurements of healthy and cancerous breast tissues that were either embedded in paraffin or deparaffinized. The resulting spectrum and accurate analysis led to the conclusion that the appropriate measurement of the background and the elimination of peaks from the paraffin had the greatest impact on the reliability of results. Furthermore, after the accurate, detailed studies FTIR and Raman spectroscopy on samples of breast tissue that were deparaffinized or embedded in paraffin, including a complete analysis of the peak after transformation Kramers-Kröning (KK), it was found that sample preparation did not affect the result obtained by measuring the reflectance in the mid-infrared range, and that this only had a minimal effect relating to the intensity obtained by the measurement of the Raman peak. Only in special cases, when Raman spectroscopic methods are used for research to find the peculiarities of the spectra, are deparaffinization recommended, in order to attain more detailed results that could be crucial in understanding the process of carcinogenesis.

Nitrogen Cycle Evaluation (NiCE) Chip for the Simultaneous Analysis of Multiple N-Cycle Associated Genes.

PubMed

Oshiki, Mamoru; Segawa, Takahiro; Ishii, Satoshi

2018-02-02

Various microorganisms play key roles in the Nitrogen (N) cycle. Quantitative PCR (qPCR) and PCR-amplicon sequencing of the N cycle functional genes allow us to analyze the abundance and diversity of microbes responsible in the N transforming reactions in various environmental samples. However, analysis of multiple target genes can be cumbersome and expensive. PCR-independent analysis, such as metagenomics and metatranscriptomics, is useful but expensive especially when we analyze multiple samples and try to detect N cycle functional genes present at relatively low abundance. Here, we present the application of microfluidic qPCR chip technology to simultaneously quantify and prepare amplicon sequence libraries for multiple N cycle functional genes as well as taxon-specific 16S rRNA gene markers for many samples. This approach, named as N cycle evaluation (NiCE) chip, was evaluated by using DNA from pure and artificially mixed bacterial cultures and by comparing the results with those obtained by conventional qPCR and amplicon sequencing methods. Quantitative results obtained by the NiCE chip were comparable to those obtained by conventional qPCR. In addition, the NiCE chip was successfully applied to examine abundance and diversity of N cycle functional genes in wastewater samples. Although non-specific amplification was detected on the NiCE chip, this could be overcome by optimizing the primer sequences in the future. As the NiCE chip can provide high-throughput format to quantify and prepare sequence libraries for multiple N cycle functional genes, this tool should advance our ability to explore N cycling in various samples. Importance. We report a novel approach, namely Nitrogen Cycle Evaluation (NiCE) chip by using microfluidic qPCR chip technology. By sequencing the amplicons recovered from the NiCE chip, we can assess diversities of the N cycle functional genes. The NiCE chip technology is applicable to analyze the temporal dynamics of the N cycle gene transcriptions in wastewater treatment bioreactors. The NiCE chip can provide high-throughput format to quantify and prepare sequence libraries for multiple N cycle functional genes. While there is a room for future improvement, this tool should significantly advance our ability to explore the N cycle in various environmental samples. Copyright © 2018 American Society for Microbiology.

Rapid production of hollow SS316 profiles by extrusion based additive manufacturing

NASA Astrophysics Data System (ADS)

Rane, Kedarnath; Cataldo, Salvatore; Parenti, Paolo; Sbaglia, Luca; Mussi, Valerio; Annoni, Massimiliano; Giberti, Hermes; Strano, Matteo

2018-05-01

Complex shaped stainless steel tubes are often required for special purpose biomedical equipment. Nevertheless, traditional manufacturing technologies, such as extrusion, lack the ability to compete in a market of customized complex components because of associated expenses towards tooling and extrusion presses. To rapid manufacture few of such components with low cost and high precision, a new Extrusion based Additive Manufacturing (EAM) process, is proposed in this paper, and as an example, short stainless steel 316L complex shaped and sectioned tubes were prepared by EAM. Several sample parts were produced using this process; the dimensional stability, surface roughness and chemical composition of sintered samples were investigated to prove process competence. The results indicate that feedstock with a 316L particle content of 92.5 wt. % can be prepared with a sigma blade mixing, whose rheological behavior is fit for EAM. The green samples have sufficient strength to handle them for subsequent treatments. The sintered samples considerably shrunk to designed dimensions and have a homogeneous microstructure to impart mechanical strength. Whereas, maintaining comparable dimensional accuracy and chemical composition which are required for biomedical equipment still need iterations, a kinematic correction and modification in debinding cycle was proposed.

Microextraction techniques combined with capillary electrophoresis in bioanalysis.

PubMed

Kohler, Isabelle; Schappler, Julie; Rudaz, Serge

2013-01-01

Over the past two decades, many environmentally sustainable sample-preparation techniques have been proposed, with the objective of reducing the use of toxic organic solvents or substituting these with environmentally friendly alternatives. Microextraction techniques (MEs), in which only a small amount of organic solvent is used, have several advantages, including reduced sample volume, analysis time, and operating costs. Thus, MEs are well adapted in bioanalysis, in which sample preparation is mandatory because of the complexity of a sample that is available in small quantities (mL or even μL only). Capillary electrophoresis (CE) is a powerful and efficient separation technique in which no organic solvents are required for analysis. Combination of CE with MEs is regarded as a very attractive environmentally sustainable analytical tool, and numerous applications have been reported over the last few decades for bioanalysis of low-molecular-weight compounds or for peptide analysis. In this paper we review the use of MEs combined with CE in bioanalysis. The review is divided into two sections: liquid and solid-based MEs. A brief practical and theoretical description of each ME is given, and the techniques are illustrated by relevant applications.

Polysialylated N-Glycans Identified in Human Serum Through Combined Developments in Sample Preparation, Separations and Electrospray ionization-mass spectrometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kronewitter, Scott R.; Marginean, Ioan; Cox, Jonathan T.

The N-glycan diversity of human serum glycoproteins, i.e. the human blood serum N-glycome, is complex due to the range of glycan structures potentially synthesizable by human glycosylation enzymes. The reported glycome, however, is limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. In this report, several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to provide an improved view of glycan diversity. Sample preparation improvements include acidified, microwave-accelerated, PNGase F N-glycan release, and sodium borohydride reduction were optimized to improvemore » quantitative yields and conserve the number of glycoforms detected. Two-stage desalting (during solid phase extraction and on the analytical column) increased the sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. On-line separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient which provides additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) has been utilized. When method improvements are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described1 these technologies demonstrate the ability to significantly extend glycan detection sensitivity and provide expanded glycan coverage. We demonstrate application of these advances in the context of the human serum glycome, and for which our initial observations include detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans.« less

Enhanced electrochemical performance of amorphous carbon nanotube-manganese-di-oxide-poly-pyrrole ternary nanohybrid

NASA Astrophysics Data System (ADS)

Pahari, D.; Das, N. S.; Das, B.; Howli, P.; Chattopadhyay, K. K.; Banerjee, D.

2017-12-01

Amorphous carbon nanotubes (a-CNTs) manganese di oxide (MnO2)-poly pyrrole (PPy) ternary nanocomposites have been synthesized by a simple chemical route. The as prepared samples have been characterized with different characterization tools that include field emission scanning and high resolution transmission electron microscopy, Raman, Fourier transformed infrared as well as UV-Vis spectroscopy. The electrochemical performance of all the as prepared pure and hybrid samples have been studied in detail. It has been seen that the ternary hybrid shows efficient electrochemical performance with high value of specific capacitance with good stability even up to 2000 cycles. The superior performance of the hybrid samples can be attributed to the strong synergistic effect between the components resulting electron shuttling along PPy main chains and inter-chain raising built-in continuous conductive network. The ternary composite approach offers an effective solution to enhance the device performance of metal-oxide based supercapacitors for long cycling applications. These studies can well speculate the existence of another supercapacitor hybrid for the use in environment friendly electrode and thus a pollution free nature.

Application of a Stir Bar Sorptive Extraction sample preparation method with HPLC for soil fungal biomass determination in soils from a detrital manipulation study.

PubMed

Beni, Áron; Lajtha, Kate; Kozma, János; Fekete, István

2017-05-01

Ergosterol is a sterol found ubiquitously in cell membranes of filamentous fungi. Although concentrations in different fungal species span the range of 2.6 to 42μg/mL of dry mass, many studies have shown a strong correlation between soil ergosterol content and fungal biomass. The analysis of ergosterol in soil therefore could be an effective tool for monitoring changes in fungal biomass under different environmental conditions. Stir Bar Sorptive Extraction (SBSE) is a new sample preparation method to extract and concentrate organic analytes from liquid samples. SBSE was here demonstrated to be a simple, fast, and cost effective method for the quantitative analysis of ergosterol from field-collected soils. Using this method we observed that soil ergosterol as a measure of fungal biomass proved to be a sensitive indicator of soil microbial dynamics that were altered by changes in plant detrital inputs to soils in a long-term field experiment. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantifying cannabis: A field study of marijuana quantity estimation.

PubMed

Prince, Mark A; Conner, Bradley T; Pearson, Matthew R

2018-06-01

The assessment of marijuana use quantity poses unique challenges. These challenges have limited research efforts on quantity assessments. However, quantity estimates are critical to detecting associations between marijuana use and outcomes. We examined accuracy of marijuana users' estimations of quantities of marijuana they prepared to ingest and predictors of both how much was prepared for a single dose and the degree of (in)accuracy of participants' estimates. We recruited a sample of 128 regular-to-heavy marijuana users for a field study wherein they prepared and estimated quantities of marijuana flower in a joint or a bowl as well as marijuana concentrate using a dab tool. The vast majority of participants overestimated the quantity of marijuana that they used in their preparations. We failed to find robust predictors of estimation accuracy. Self-reported quantity estimates are inaccurate, which has implications for studying the link between quantity and marijuana use outcomes. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples.

PubMed

Lantz, P G; Abu al-Soud, W; Knutsson, R; Hahn-Hägerdal, B; Rådström, P

2000-01-01

Since its introduction in the mid-80s, polymerase chain reaction (PCR) technology has been recognised as a rapid, sensitive and specific molecular diagnostic tool for the analysis of micro-organisms in clinical, environmental and food samples. Although this technique can be extremely effective with pure solutions of nucleic acids, it's sensitivity may be reduced dramatically when applied directly to biological samples. This review describes PCR technology as a microbial detection method, PCR inhibitors in biological samples and various sample preparation techniques that can be used to facilitate PCR detection, by either separating the micro-organisms from PCR inhibitors and/or by concentrating the micro-organisms to detectable concentrations. Parts of this review are updated and based on a doctoral thesis by Lantz [1] and on a review discussing methods to overcome PCR inhibition in foods [2].

Anisotropic propagation imaging of elastic waves in oriented columnar thin films

NASA Astrophysics Data System (ADS)

Coffy, E.; Dodane, G.; Euphrasie, S.; Mosset, A.; Vairac, P.; Martin, N.; Baida, H.; Rampnoux, J. M.; Dilhaire, S.

2017-12-01

We report on the observation of strongly anisotropic surface acoustic wave propagation on nanostructured thin films. Two kinds of tungsten samples were prepared by sputtering on a silicon substrate: a conventional thin film with columns normal to the substrate surface, and an oriented columnar architecture using the glancing angle deposition (GLAD) process. Pseudo-Rayleigh waves (PRWs) were imaged as a function of time in x and y directions for both films thanks to a femtosecond heterodyne pump-probe setup. A strong anisotropic propagation as well as a high velocity reduction of the PRWs were exhibited for the GLAD sample. For the wavevector k/2π  =  3  ×  105 m-1 the measured group velocities v x and v y equal 2220 m s-1 for the sample prepared with conventional sputtering, whereas a strong anisotropy appears (v x   =  1600 m s-1 and v y   =  870 m s-1) for the sample prepared with the GLAD process. Using the finite element method, the anisotropy is related to the structural anisotropy of the thin film’s architecture. The drop of PRWs group velocities is mainly assigned to the porous microstructure, especially favored by atomic shadowing effects which appear during the growth of the inclined columns. Such GLAD thin films constitute a new tool for the control of the propagation of surface elastic waves and for the design of new devices with useful properties.

Conversion and Retrievability of Hard Copy and Digital Documents on Optical Disks

DTIC Science & Technology

1992-03-01

53 B. CURRENT THESIS PREPARATION TOOLS . ....... 54 1. Thesis Preparation using G-Thesis ..... 55 2. Thesis Preparation using Framemaker ...School mainframe. • Computer Science department students can use a software package called Framemaker , available on Sun work stations in their...by most thesis typists and students. For this reason, the discussion of thesis preparation tools will be limited to; G-thesis, Framemaker and

[Progress in the application of laser ablation ICP-MS to surface microanalysis in material science].

PubMed

Zhang, Yong; Jia, Yun-hai; Chen, Ji-wen; Shen, Xue-jing; Liu, Ying; Zhao, Leiz; Li, Dong-ling; Hang, Peng-cheng; Zhao, Zhen; Fan, Wan-lun; Wang, Hai-zhou

2014-08-01

In the present paper, apparatus and theory of surface analysis is introduced, and the progress in the application of laser ablation ICP-MS to microanalysis in ferrous, nonferrous and semiconductor field is reviewed in detail. Compared with traditional surface analytical tools, such as SEM/EDS (scanning electron microscopy/energy dispersive spectrum), EPMA (electron probe microanalysis analysis), AES (auger energy spectrum), etc. the advantage is little or no sample preparation, adjustable spatial resolution according to analytical demand, multi-element analysis and high sensitivity. It is now a powerful complementary method to traditional surface analytical tool. With the development of LA-ICP-MS technology maturing, more and more analytical workers will use this powerful tool in the future, and LA-ICP-MS will be a super star in elemental analysis field just like LIBS (Laser-induced breakdown spectroscopy).

Evaluation of a gas chromatography method for azelaic acid determination in selected biological samples

PubMed Central

Garelnabi, Mahdi; Litvinov, Dmitry; Parthasarathy, Sampath

2010-01-01

Background: Azelaic acid (AzA) is the best known dicarboxilic acid to have pharmaceutical benefits and clinical applications and also to be associated with some diseases pathophysiology. Materials and Methods: We extracted and methylesterified AzA and determined its concentration in human plasma obtained from healthy individuals and also in mice fed AzA containing diet for three months. Results: AzA was detected in Gas Chromatography (GC) and confirmed by Liquid chromatography mass spectrometry (LCMS), and gas chromatography mass spectrometry (GCMC). Our results have shown that AzA can be determined efficiently in selected biological samples by GC method with 1nM limit of detection (LoD) and the limit of quantification (LoQ); was established at 50nM. Analytical Sensitivity as assayed by hexane demonstrated an analytical sensitivity at 0.050nM. The method has demonstrated 8-10% CV batch repeatability across the sample types and 13-18.9% CV for the Within-Lab Precision analysis. The method has shown that AzA can efficiently be recovered from various sample preparation including liver tissue homogenate (95%) and human plasma (97%). Conclusions: Because of its simplicity and lower limit of quantification, the present method provides a useful tool for determining AzA in various biological sample preparations. PMID:22558586

Evaluation of a gas chromatography method for azelaic acid determination in selected biological samples.

PubMed

Garelnabi, Mahdi; Litvinov, Dmitry; Parthasarathy, Sampath

2010-09-01

Azelaic acid (AzA) is the best known dicarboxilic acid to have pharmaceutical benefits and clinical applications and also to be associated with some diseases pathophysiology. We extracted and methylesterified AzA and determined its concentration in human plasma obtained from healthy individuals and also in mice fed AzA containing diet for three months. AzA was detected in Gas Chromatography (GC) and confirmed by Liquid chromatography mass spectrometry (LCMS), and gas chromatography mass spectrometry (GCMC). Our results have shown that AzA can be determined efficiently in selected biological samples by GC method with 1nM limit of detection (LoD) and the limit of quantification (LoQ); was established at 50nM. Analytical Sensitivity as assayed by hexane demonstrated an analytical sensitivity at 0.050nM. The method has demonstrated 8-10% CV batch repeatability across the sample types and 13-18.9% CV for the Within-Lab Precision analysis. The method has shown that AzA can efficiently be recovered from various sample preparation including liver tissue homogenate (95%) and human plasma (97%). Because of its simplicity and lower limit of quantification, the present method provides a useful tool for determining AzA in various biological sample preparations.

Sex Differences in Object Manipulation in Wild Immature Chimpanzees (Pan troglodytes schweinfurthii) and Bonobos (Pan paniscus): Preparation for Tool Use?

PubMed

Koops, Kathelijne; Furuichi, Takeshi; Hashimoto, Chie; van Schaik, Carel P

2015-01-01

Sex differences in immatures predict behavioural differences in adulthood in many mammal species. Because most studies have focused on sex differences in social interactions, little is known about possible sex differences in 'preparation' for adult life with regards to tool use skills. We investigated sex and age differences in object manipulation in immature apes. Chimpanzees use a variety of tools across numerous contexts, whereas bonobos use few tools and none in foraging. In both species, a female bias in adult tool use has been reported. We studied object manipulation in immature chimpanzees at Kalinzu (Uganda) and bonobos at Wamba (Democratic Republic of Congo). We tested predictions of the 'preparation for tool use' hypothesis. We confirmed that chimpanzees showed higher rates and more diverse types of object manipulation than bonobos. Against expectation, male chimpanzees showed higher object manipulation rates than females, whereas in bonobos no sex difference was found. However, object manipulation by male chimpanzees was play-dominated, whereas manipulation types of female chimpanzees were more diverse (e.g., bite, break, carry). Manipulation by young immatures of both species was similarly dominated by play, but only in chimpanzees did it become more diverse with age. Moreover, in chimpanzees, object types became more tool-like (i.e., sticks) with age, further suggesting preparation for tool use in adulthood. The male bias in object manipulation in immature chimpanzees, along with the late onset of tool-like object manipulation, indicates that not all (early) object manipulation (i.e., object play) in immatures prepares for subsistence tool use. Instead, given the similarity with gender differences in human children, object play may also function in motor skill practice for male-specific behaviours (e.g., dominance displays). In conclusion, even though immature behaviours almost certainly reflect preparation for adult roles, more detailed future work is needed to disentangle possible functions of object manipulation during development.

Integration of gas chromatography mass spectrometry methods for differentiating ricin preparation methods.

PubMed

Wunschel, David S; Melville, Angela M; Ehrhardt, Christopher J; Colburn, Heather A; Victry, Kristin D; Antolick, Kathryn C; Wahl, Jon H; Wahl, Karen L

2012-05-07

The investigation of crimes involving chemical or biological agents is infrequent, but presents unique analytical challenges. The protein toxin ricin is encountered more frequently than other agents and is found in the seeds of Ricinus communis, commonly known as the castor plant. Typically, the toxin is extracted from castor seeds utilizing a variety of different recipes that result in varying purity of the toxin. Moreover, these various purification steps can also leave or differentially remove a variety of exogenous and endogenous residual components with the toxin that may indicate the type and number of purification steps involved. We have applied three gas chromatography-mass spectrometry (GC-MS) based analytical methods to measure the variation in seed carbohydrates and castor oil ricinoleic acid, as well as the presence of solvents used for purification. These methods were applied to the same samples prepared using four previously identified toxin preparation methods, starting from four varieties of castor seeds. The individual data sets for seed carbohydrate profiles, ricinoleic acid, or acetone amount each provided information capable of differentiating different types of toxin preparations across seed types. However, the integration of the data sets using multivariate factor analysis provided a clear distinction of all samples based on the preparation method, independent of the seed source. In particular, the abundance of mannose, arabinose, fucose, ricinoleic acid, and acetone were shown to be important differentiating factors. These complementary tools provide a more confident determination of the method of toxin preparation than would be possible using a single analytical method.

Using populations of human and microbial genomes for organism detection in metagenomes

DOE PAGES

Ames, Sasha K.; Gardner, Shea N.; Marti, Jose Manuel; ...

2015-04-29

Identifying causative disease agents in human patients from shotgun metagenomic sequencing (SMS) presents a powerful tool to apply when other targeted diagnostics fail. Numerous technical challenges remain, however, before SMS can move beyond the role of research tool. Accurately separating the known and unknown organism content remains difficult, particularly when SMS is applied as a last resort. The true amount of human DNA that remains in a sample after screening against the human reference genome and filtering nonbiological components left from library preparation has previously been underreported. In this study, we create the most comprehensive collection of microbial and reference-freemore » human genetic variation available in a database optimized for efficient metagenomic search by extracting sequences from GenBank and the 1000 Genomes Project. The results reveal new human sequences found in individual Human Microbiome Project (HMP) samples. Individual samples contain up to 95% human sequence, and 4% of the individual HMP samples contain 10% or more human reads. In conclusion, left unidentified, human reads can complicate and slow down further analysis and lead to inaccurately labeled microbial taxa and ultimately lead to privacy concerns as more human genome data is collected.« less

Using populations of human and microbial genomes for organism detection in metagenomes.

PubMed

Ames, Sasha K; Gardner, Shea N; Marti, Jose Manuel; Slezak, Tom R; Gokhale, Maya B; Allen, Jonathan E

2015-07-01

Identifying causative disease agents in human patients from shotgun metagenomic sequencing (SMS) presents a powerful tool to apply when other targeted diagnostics fail. Numerous technical challenges remain, however, before SMS can move beyond the role of research tool. Accurately separating the known and unknown organism content remains difficult, particularly when SMS is applied as a last resort. The true amount of human DNA that remains in a sample after screening against the human reference genome and filtering nonbiological components left from library preparation has previously been underreported. In this study, we create the most comprehensive collection of microbial and reference-free human genetic variation available in a database optimized for efficient metagenomic search by extracting sequences from GenBank and the 1000 Genomes Project. The results reveal new human sequences found in individual Human Microbiome Project (HMP) samples. Individual samples contain up to 95% human sequence, and 4% of the individual HMP samples contain 10% or more human reads. Left unidentified, human reads can complicate and slow down further analysis and lead to inaccurately labeled microbial taxa and ultimately lead to privacy concerns as more human genome data is collected. © 2015 Ames et al.; Published by Cold Spring Harbor Laboratory Press.

Using populations of human and microbial genomes for organism detection in metagenomes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ames, Sasha K.; Gardner, Shea N.; Marti, Jose Manuel

Identifying causative disease agents in human patients from shotgun metagenomic sequencing (SMS) presents a powerful tool to apply when other targeted diagnostics fail. Numerous technical challenges remain, however, before SMS can move beyond the role of research tool. Accurately separating the known and unknown organism content remains difficult, particularly when SMS is applied as a last resort. The true amount of human DNA that remains in a sample after screening against the human reference genome and filtering nonbiological components left from library preparation has previously been underreported. In this study, we create the most comprehensive collection of microbial and reference-freemore » human genetic variation available in a database optimized for efficient metagenomic search by extracting sequences from GenBank and the 1000 Genomes Project. The results reveal new human sequences found in individual Human Microbiome Project (HMP) samples. Individual samples contain up to 95% human sequence, and 4% of the individual HMP samples contain 10% or more human reads. In conclusion, left unidentified, human reads can complicate and slow down further analysis and lead to inaccurately labeled microbial taxa and ultimately lead to privacy concerns as more human genome data is collected.« less

Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples

PubMed Central

Quick, Josh; Grubaugh, Nathan D; Pullan, Steven T; Claro, Ingra M; Smith, Andrew D; Gangavarapu, Karthik; Oliveira, Glenn; Robles-Sikisaka, Refugio; Rogers, Thomas F; Beutler, Nathan A; Burton, Dennis R; Lewis-Ximenez, Lia Laura; de Jesus, Jaqueline Goes; Giovanetti, Marta; Hill, Sarah; Black, Allison; Bedford, Trevor; Carroll, Miles W; Nunes, Marcio; Alcantara, Luiz Carlos; Sabino, Ester C; Baylis, Sally A; Faria, Nuno; Loose, Matthew; Simpson, Jared T; Pybus, Oliver G; Andersen, Kristian G; Loman, Nicholas J

2018-01-01

Genome sequencing has become a powerful tool for studying emerging infectious diseases; however, genome sequencing directly from clinical samples without isolation remains challenging for viruses such as Zika, where metagenomic sequencing methods may generate insufficient numbers of viral reads. Here we present a protocol for generating coding-sequence complete genomes comprising an online primer design tool, a novel multiplex PCR enrichment protocol, optimised library preparation methods for the portable MinION sequencer (Oxford Nanopore Technologies) and the Illumina range of instruments, and a bioinformatics pipeline for generating consensus sequences. The MinION protocol does not require an internet connection for analysis, making it suitable for field applications with limited connectivity. Our method relies on multiplex PCR for targeted enrichment of viral genomes from samples containing as few as 50 genome copies per reaction. Viral consensus sequences can be achieved starting with clinical samples in 1-2 days following a simple laboratory workflow. This method has been successfully used by several groups studying Zika virus evolution and is facilitating an understanding of the spread of the virus in the Americas. PMID:28538739

Apes produce tools for future use.

PubMed

Bräuer, Juliane; Call, Josep

2015-03-01

There is now growing evidence that some animal species are able to plan for the future. For example great apes save and exchange tools for future use. Here we raise the question whether chimpanzees, orangutans, and bonobos would produce tools for future use. Subjects only had access to a baited apparatus for a limited duration and therefore should use the time preceding this access to create the appropriate tools in order to get the rewards. The apes were tested in three conditions depending on the need for pre-prepared tools. Either eight tools, one tool or no tools were needed to retrieve the reward. The apes prepared tools in advance for future use and they produced them mainly in conditions when they were really needed. The fact that apes were able to solve this new task indicates that their planning skills are flexible. However, for the condition in which eight tools were needed, apes produced less than two tools per trial in advance. However, they used their chance to produce additional tools in the tool use phase-thus often obtaining most of the reward from the apparatus. Increased pressure to prepare more tools in advance did not have an effect on their performance. © 2014 Wiley Periodicals, Inc.

Advances in Software Tools for Pre-processing and Post-processing of Overset Grid Computations

NASA Technical Reports Server (NTRS)

Chan, William M.

2004-01-01

Recent developments in three pieces of software for performing pre-processing and post-processing work on numerical computations using overset grids are presented. The first is the OVERGRID graphical interface which provides a unified environment for the visualization, manipulation, generation and diagnostics of geometry and grids. Modules are also available for automatic boundary conditions detection, flow solver input preparation, multiple component dynamics input preparation and dynamics animation, simple solution viewing for moving components, and debris trajectory analysis input preparation. The second is a grid generation script library that enables rapid creation of grid generation scripts. A sample of recent applications will be described. The third is the OVERPLOT graphical interface for displaying and analyzing history files generated by the flow solver. Data displayed include residuals, component forces and moments, number of supersonic and reverse flow points, and various dynamics parameters.

Preparation of herpes simplex virus-infected primary neurons for transmission electron microscopy.

PubMed

Miranda-Saksena, Monica; Boadle, Ross; Cunningham, Anthony L

2014-01-01

Transmission electron microscopy (TEM) provides the resolution necessary to identify both viruses and subcellular components of cells infected with many types of viruses, including herpes simplex virus. Recognized as a powerful tool in both diagnostic and research-based virology laboratories, TEM has made possible the identification of new viruses and has contributed to the elucidation of virus life cycle and virus-host cell interaction. Whilst there are many sample preparation techniques for TEM, conventional processing using chemical fixation and resin embedding remains a useful technique, available in virtually all EM laboratories, for studying virus/cell ultrastructure. In this chapter, we describe the preparation of herpes simplex virus-infected primary neurons, grown on plastic cover slips, to allow sectioning of neurons and axons in their growth plane. This technique allows TEM examination of cell bodies, axons, growth cones, and varicosities, providing powerful insights into virus-cell interaction.

Stability of dry coated solid dosage forms.

PubMed

Kablitz, Caroline Désirée; Urbanetz, Nora Anne

2009-01-01

The dry coating process was evaluated in terms of storage stability investigating drug release and agglomeration tendency of the different coated oral dosage forms; hydroxypropyl methylcellulose acetate succinate (HPMCAS) was used with triethylcitrate (TEC) as plasticizer and acetylated monoglyceride (Myvacet) as wetting agent. Talc or colloidal silicon dioxide (Aerosil) was used as anti-tacking agents. In contrast to coating formulations consisting of HPMCAS and Myvacet all formulations containing TEC showed enteric resistance and no agglomeration tendency after preparation. After storage at 10% RH +/- 5% enteric resistance is increased slightly. This increase is more pronounced at 60% RH +/- 5%. The formulations without anti-tacking agents showed higher drug releases after 12 and 24 months due to the damage of the film's integrity during sample preparation caused by the high tackiness of the film. Tackiness is not affected by storing if samples are stored at low relative humidity. At high relative humidity tackiness increases upon storage especially for formulations without anti-tacking agents. The sieving results of the agglomeration measurements after storage can be confirmed by ring shear measurements performed immediately after preparation and approved to be a tool, which is able to predict the agglomeration during storage.

Treatment Cost Analysis Tool (TCAT) for Estimating Costs of Outpatient Treatment Services

PubMed Central

Flynn, Patrick M.; Broome, Kirk M.; Beaston-Blaakman, Aaron; Knight, Danica K.; Horgan, Constance M.; Shepard, Donald S.

2009-01-01

A Microsoft® Excel-based workbook designed for research analysts to use in a national study was retooled for treatment program directors and financial officers to allocate, analyze, and estimate outpatient treatment costs in the U.S. This instrument can also be used as a planning and management tool to optimize resources and forecast the impact of future changes in staffing, client flow, program design, and other resources. The Treatment Cost Analysis Tool (TCAT) automatically provides feedback and generates summaries and charts using comparative data from a national sample of non-methadone outpatient providers. TCAT is being used by program staff to capture and allocate both economic and accounting costs, and outpatient service costs are reported for a sample of 70 programs. Costs for an episode of treatment in regular, intensive, and mixed types of outpatient treatment types were $882, $1,310, and $1,381 respectively (based on 20% trimmed means and 2006 dollars). An hour of counseling cost $64 in regular, $85 intensive, and $86 mixed. Group counseling hourly costs per client were $8, $11, and $10 respectively for regular, intensive, and mixed. Future directions include use of a web-based interview version, much like some of the commercially available tax preparation software tools, and extensions for use in other modalities of treatment. PMID:19004576

Treatment Cost Analysis Tool (TCAT) for estimating costs of outpatient treatment services.

PubMed

Flynn, Patrick M; Broome, Kirk M; Beaston-Blaakman, Aaron; Knight, Danica K; Horgan, Constance M; Shepard, Donald S

2009-02-01

A Microsoft Excel-based workbook designed for research analysts to use in a national study was retooled for treatment program directors and financial officers to allocate, analyze, and estimate outpatient treatment costs in the U.S. This instrument can also be used as a planning and management tool to optimize resources and forecast the impact of future changes in staffing, client flow, program design, and other resources. The Treatment Cost Analysis Tool (TCAT) automatically provides feedback and generates summaries and charts using comparative data from a national sample of non-methadone outpatient providers. TCAT is being used by program staff to capture and allocate both economic and accounting costs, and outpatient service costs are reported for a sample of 70 programs. Costs for an episode of treatment in regular, intensive, and mixed types of outpatient treatment were $882, $1310, and $1381 respectively (based on 20% trimmed means and 2006 dollars). An hour of counseling cost $64 in regular, $85 intensive, and $86 mixed. Group counseling hourly costs per client were $8, $11, and $10 respectively for regular, intensive, and mixed. Future directions include use of a web-based interview version, much like some of the commercially available tax preparation software tools, and extensions for use in other modalities of treatment.

Determination of renewable energy yield from mixed waste material from the use of novel image analysis methods.

PubMed

Wagland, S T; Dudley, R; Naftaly, M; Longhurst, P J

2013-11-01

Two novel techniques are presented in this study which together aim to provide a system able to determine the renewable energy potential of mixed waste materials. An image analysis tool was applied to two waste samples prepared using known quantities of source-segregated recyclable materials. The technique was used to determine the composition of the wastes, where through the use of waste component properties the biogenic content of the samples was calculated. The percentage renewable energy determined by image analysis for each sample was accurate to within 5% of the actual values calculated. Microwave-based multiple-point imaging (AutoHarvest) was used to demonstrate the ability of such a technique to determine the moisture content of mixed samples. This proof-of-concept experiment was shown to produce moisture measurement accurate to within 10%. Overall, the image analysis tool was able to determine the renewable energy potential of the mixed samples, and the AutoHarvest should enable the net calorific value calculations through the provision of moisture content measurements. The proposed system is suitable for combustion facilities, and enables the operator to understand the renewable energy potential of the waste prior to combustion. Copyright © 2013 Elsevier Ltd. All rights reserved.

Video Games as a Training Tool to Prepare the Next Generation of Cyber Warriors

DTIC Science & Technology

2014-10-01

2. REPORT TYPE N/A 3. DATES COVERED - 4 . TITLE AND SUBTITLE Video Games as a Training Tool to Prepare the Next Generation of Cyber Warriors...CYBERSECURITY WORKFORCE SHORTAGE .......................................................................... 3 4 1.1 GREATER CYBERSECURITY EDUCATION IS... 4 6 2.1 HOW VIDEO GAMES CAN BE EFFECTIVE LEARNING TOOLS

Application of Microextraction Techniques Including SPME and MESI to the Thermal Degradation of Polymers: A Review.

PubMed

Kaykhaii, Massoud; Linford, Matthew R

2017-03-04

Here, we discuss the newly developed micro and solventless sample preparation techniques SPME (Solid Phase Microextraction) and MESI (Membrane Extraction with a Sorbent Interface) as applied to the qualitative and quantitative analysis of thermal oxidative degradation products of polymers and their stabilizers. The coupling of these systems to analytical instruments is also described. Our comprehensive literature search revealed that there is no previously published review article on this topic. It is shown that these extraction techniques are valuable sample preparation tools for identifying complex series of degradation products in polymers. In general, the number of products identified by traditional headspace (HS-GC-MS) is much lower than with SPME-GC-MS. MESI is particularly well suited for the detection of non-polar compounds, therefore number of products identified by this technique is not also to the same degree of SPME. Its main advantage, however, is its ability of (semi-) continuous monitoring, but it is more expensive and not yet commercialized.

"Soft"or "hard" ionisation? Investigation of metastable gas temperature effect on direct analysis in real-time analysis of Voriconazole.

PubMed

Lapthorn, Cris; Pullen, Frank

2009-01-01

The performance of the direct analysis in real-time (DART) technique was evaluated across a range of metastable gas temperatures for a pharmaceutical compound, Voriconazole, in order to investigate the effect of metastable gas temperature on molecular ion intensity and fragmentation. The DART source has been used to analyse a range of analytes and from a range of matrices including drugs in solid tablet form and preparations, active ingredients in ointment, naturally occurring plant alkaloids, flavours and fragrances, from thin layer chromatography (TLC) plates, melting point tubes and biological matrices including hair, urine and blood. The advantages of this technique include rapid analysis time (as little as 5 s), a reduction in sample preparation requirements, elimination of mobile phase requirement and analysis of samples not typically amenable to atmospheric pressure ionisation (API) techniques. This technology has therefore been proposed as an everyday tool for identification of components in crude organic reaction mixtures.

RNA-Seq for Bacterial Gene Expression.

PubMed

Poulsen, Line Dahl; Vinther, Jeppe

2018-06-01

RNA sequencing (RNA-seq) has become the preferred method for global quantification of bacterial gene expression. With the continued improvements in sequencing technology and data analysis tools, the most labor-intensive and expensive part of an RNA-seq experiment is the preparation of sequencing libraries, which is also essential for the quality of the data obtained. Here, we present a straightforward and inexpensive basic protocol for preparation of strand-specific RNA-seq libraries from bacterial RNA as well as a computational pipeline for the data analysis of sequencing reads. The protocol is based on the Illumina platform and allows easy multiplexing of samples and the removal of sequencing reads that are PCR duplicates. © 2018 by John Wiley & Sons, Inc. © 2018 John Wiley & Sons, Inc.

Fabrication and Characterization of High Temperature Resin/Carbon Nanofiber Composites

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Working, Dennis C.; Criss, Jim M.; Siochi, Emilie J.; Conell, John W.

2005-01-01

As part of ongoing efforts to develop multifunctional advanced composites, blends of PETI-330 and carbon nanofibers (CNF) were prepared and characterized. Dry mixing techniques were employed and the effect of CNF loading level on melt viscosity was determined. The resulting powders were characterized for degree of mixing, thermal and rheological properties. Based on the characterization results, samples containing 30 and 40 wt% CNF were scaled up to approx.300 g and used to fabricate moldings 10.2 cm x 15.2 cm x 0.32 cm thick. The moldings were fabricated by injecting the mixtures at 260-280 C into a stainless steel tool followed by curing for 1 h at 371 C. The tool was designed to impart high shear during the injection process in an attempt to achieve some alignment of CNFs in the flow direction. Moldings were obtained that were subsequently characterized for thermal, mechanical and electrical properties. The degree of dispersion and alignment of CNFs were investigated using high-resolution scanning electron microscopy. The preparation and preliminary characterization of PETI-330/CNF composites will be discussed.

Probing the underlying physics of ejecta production from shocked Sn samples

NASA Astrophysics Data System (ADS)

Zellner, M. B.; McNeil, W. Vogan; Hammerberg, J. E.; Hixson, R. S.; Obst, A. W.; Olson, R. T.; Payton, J. R.; Rigg, P. A.; Routley, N.; Stevens, G. D.; Turley, W. D.; Veeser, L.; Buttler, W. T.

2008-06-01

This effort investigates the underlying physics of ejecta production for high explosive (HE) shocked Sn surfaces prepared with finishes typical to those roughened by tool marks left from machining processes. To investigate the physical mechanisms of ejecta production, we compiled and re-examined ejecta data from two experimental campaigns [W. S. Vogan et al., J. Appl. Phys. 98, 113508 (1998); M. B. Zellner et al., ibid. 102, 013522 (2007)] to form a self-consistent data set spanning a large parameter space. In the first campaign, ejecta created upon shock release at the back side of HE shocked Sn samples were characterized for samples with varying surface finishes but at similar shock-breakout pressures PSB. In the second campaign, ejecta were characterized for HE shocked Sn samples with a constant surface finish but at varying PSB.

doGlycans-Tools for Preparing Carbohydrate Structures for Atomistic Simulations of Glycoproteins, Glycolipids, and Carbohydrate Polymers for GROMACS.

PubMed

Danne, Reinis; Poojari, Chetan; Martinez-Seara, Hector; Rissanen, Sami; Lolicato, Fabio; Róg, Tomasz; Vattulainen, Ilpo

2017-10-23

Carbohydrates constitute a structurally and functionally diverse group of biological molecules and macromolecules. In cells they are involved in, e.g., energy storage, signaling, and cell-cell recognition. All of these phenomena take place in atomistic scales, thus atomistic simulation would be the method of choice to explore how carbohydrates function. However, the progress in the field is limited by the lack of appropriate tools for preparing carbohydrate structures and related topology files for the simulation models. Here we present tools that fill this gap. Applications where the tools discussed in this paper are particularly useful include, among others, the preparation of structures for glycolipids, nanocellulose, and glycans linked to glycoproteins. The molecular structures and simulation files generated by the tools are compatible with GROMACS.

Fusion Bead Procedure for Nuclear Forensics Employing Synthetic Enstatite to Dissolve Uraniferous and Other Challenging Materials Prior to Laser Ablation Inductively Coupled Plasma Mass Spectrometry.

PubMed

Reading, David G; Croudace, Ian W; Warwick, Phillip E

2017-06-06

There is an increasing demand for rapid and effective analytical tools to support nuclear forensic investigations of seized or suspect materials. Some methods are simply adapted from other scientific disciplines and can effectively be used to rapidly prepare complex materials for subsequent analysis. A novel sample fusion method is developed, tested, and validated to produce homogeneous, flux-free glass beads of geochemical reference materials (GRMs), uranium ores, and uranium ore concentrates (UOC) prior to the analysis of 14 rare earth elements (REE) via laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The novelty of the procedure is the production of glass beads using 9 parts high purity synthetic enstatite (MgSiO 3 ) as the glass former with 1 part of sample (sample mass ∼1.5 mg). The beads are rapidly prepared (∼10 min overall time) by fusing the blended mixture on an iridium strip resistance heater in an argon-purged chamber. Many elements can be measured in the glass bead, but the rare earth group in particular is a valuable series in nuclear forensic studies and is well-determined using LA-ICP-MS. The REE data obtained from the GRMs, presented as chondrite normalized patterns, are in very good agreement with consensus patterns. The UOCs have comparable patterns to solution ICP-MS methods and published data. The attractions of the current development are its conservation of sample, speed of preparation, and suitability for microbeam analysis, all of which are favorable for nuclear forensics practitioners and geochemists requiring REE patterns from scarce or valuable samples.

The DISAM Journal of International Security Assistance Management. Volume 24, Number 4, Summer 2002

DTIC Science & Technology

2002-01-01

to the SAFR-hosted Holiday Party where they prepare favorite dishes to be sampled and savored. While they are in the U.S., they take advantage of the...tool facilitates the generation of a multitude of reports providing insight into trends and the analysis of business processes and case management...nations, Canada, Japan, Australia, and New Zealand. A listing of countries located in regions defined for the purpose of this analysis –Asia, Near East

Computer program to assess impact of fatigue and fracture criteria on weight and cost of transport aircraft

NASA Technical Reports Server (NTRS)

Tanner, C. J.; Kruse, G. S.; Oman, B. H.

1975-01-01

A preliminary design analysis tool for rapidly performing trade-off studies involving fatigue, fracture, static strength, weight, and cost is presented. Analysis subprograms were developed for fatigue life, crack growth life, and residual strength; and linked to a structural synthesis module which in turn was integrated into a computer program. The part definition module of a cost and weight analysis program was expanded to be compatible with the upgraded structural synthesis capability. The resultant vehicle design and evaluation program is named VDEP-2. It is an accurate and useful tool for estimating purposes at the preliminary design stage of airframe development. A sample case along with an explanation of program applications and input preparation is presented.

Monitoring Interfacial Lipid Oxidation in Oil-in-Water Emulsions Using Spatially Resolved Optical Techniques.

PubMed

Banerjee, Chiranjib; Westberg, Michael; Breitenbach, Thomas; Bregnhøj, Mikkel; Ogilby, Peter R

2017-06-06

The oxidation of lipids is an important phenomenon with ramifications for disciplines that range from food science to cell biology. The development and characterization of tools and techniques to monitor lipid oxidation are thus relevant. Of particular significance in this regard are tools that facilitate the study of oxidations at interfaces in heterogeneous samples (e.g., oil-in-water emulsions, cell membranes). In this article, we establish a proof-of-principle for methods to initiate and then monitor such oxidations with high spatial resolution. The experiments were performed using oil-in-water emulsions of polyunsaturated fatty acids (PUFAs) prepared from cod liver oil. We produced singlet oxygen at a point near the oil-water interface of a given PUFA droplet in a spatially localized two-photon photosensitized process. We then followed the oxidation reactions initiated by this process with the fluorescence-based imaging technique of structured illumination microscopy (SIM). We conclude that the approach reported herein has attributes well-suited to the study of lipid oxidation in heterogeneous samples.

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maddi, Balakrishna; Panisko, Ellen; Albrecht, Karl

Two-dimensional gas chromatography coupled with time of flight mass spectrometry is a powerful tool for identifying and quantifying components in complex mixtures. It has been used to analyze gasoline, jet fuel, diesel, bio-diesel and organic fraction of bio-crude/bio-oil. In these experiments, the first dimension of separation was non-polar, followed by a polar separation. Aqueous fractions of bio-crude and other aqueous samples have been examined with similar column combinations. However, sample preparation techniques such as derivatization, solvent extraction, and solid-phase extraction were necessary prior to analysis. In this study, aqueous fraction obtained from hydrothermal liquefaction of algae was characterized by two-dimensionalmore » gas chromatography coupled with time of flight mass spectrometry without prior sample preparation techniques using a polar separation in the first dimension followed by a non-polar separation. Two-dimensional plots from this analysis were compared with those obtained from the more traditional column combination. Results from qualitative characterization aqueous fractions of algal bio-crude are discussed in detail. The advantages of using a polar separation followed by a non-polar separation for characterization of organics in aqueous samples by two-dimensional gas chromatography coupled with time of flight mass spectrometry are highlighted.« less

Surface morphology, optical, and electrochromic properties of nanostructured nickel ferrite (NiFe2O4) prepared by sol-gel method: effects of Ni/Fe molar ratios

NASA Astrophysics Data System (ADS)

Bazhan, Z.; Ghodsi, F. E.; Mazloom, J.

2016-05-01

Nanostructured nickel ferrite (NF) was prepared by the sol-gel method and calcined at 500 °C for 2 h. The effect of Ni/Fe molar ratios (0, 10, 30, 50 %) on structural, morphological, compositional, optical, and magnetic properties of samples was investigated using analytical tools. XRD patterns indicated the presence of hematite phase in the pure and 10 % NF samples. The samples of 30 and 50 % Ni/Fe molar ratios showed the formation of nickel ferrite structure. Using AFM images, power spectrum density analysis were performed for Ni/Fe with different molar ratio. Also the effect of thickness on morphology of 30 % sample was studied. The fractal dimension increases by increasing the Ni/Fe molar ratio. Optical parameters were evaluated by theoretical approach, and compositional dependence of these parameters was discussed comprehensively. Band gap narrowing was observed in nickel ferrite thin films by increasing the nickel contents from 10 to 50 %. Magnetic analysis revealed that increasing nickel content improved the saturation magnetization. Electrochemical measurements indicated that NF thin films have higher total charge density rather than Fe2O3 thin films and the ion storage capacitance of NF thin films increased by increasing the Ni/Fe content.

Label-free virus detection using silicon photonic microring resonators

PubMed Central

McClellan, Melinda S.; Domier, Leslie L; Bailey, Ryan C.

2013-01-01

Viruses represent a continual threat to humans through a number of mechanisms, which include disease, bioterrorism, and destruction of both plant and animal food resources. Many contemporary techniques used for the detection of viruses and viral infections suffer from limitations such as the need for extensive sample preparation or the lengthy window between infection and measurable immune response, for serological methods. In order to develop a method that is fast, cost-effective, and features reduced sample preparation compared to many other virus detection methods, we report the application of silicon photonic microring resonators for the direct, label-free detection of intact viruses in both purified samples as well as in a complex, real-world analytical matrix. As a model system, we demonstrate the quantitative detection of Bean pod mottle virus, a pathogen of great agricultural importance, with a limit of detection of 10 ng/mL. By simply grinding a small amount of leaf sample in buffer with a mortar and pestle, infected leaves can be identified over a healthy control with a total analysis time of less than 45 min. Given the inherent scalability and multiplexing capability of the semiconductor-based technology, we feel that silicon photonic microring resonators are well-positioned as a promising analytical tool for a number of viral detection applications. PMID:22138465

Development of a Planet Tool for an interactive School Atlas as eBook

NASA Astrophysics Data System (ADS)

Wondrak, Stephan

2018-05-01

The present thesis describes the development of a planet tool for an interactive school atlas using an eBook format. Especially the technical and cartographical capabilities of the open standard ePUB 3 are evaluated. An eBook application with interactive and dynamic 2-dimensional visualizations is developed especially to show whether the re-al-world dimensions and distances in the solar system can be mapped in a cartographical correct and for students easy understandable manner. In the first part of the work, the requirements of the planet tool are evaluated in co-operation with experts. The open standards PDF and ePUB 3 are investigated with regard to the requirements for the development of the planet tool. Another chapter describes in detail all significant steps of the development process for a prototype of the planet tool. A graphic file originally created for print production is prepared and enhanced with interactive features to generate one of the eBook pages. This serves to show a potential workflow for the generation of eBook pages in a cross-media atlas production. All sample pages of the prototype show different layouts and contain the entire spectrum of interactive features and multimedia content of modern eBooks. The sample pages are presented and discussed in an own chapter. The results of the present work aim at answering the question concerning the suitability of the open standard ePUB 3 for the development of a multimedia eBook for high school education.

Wiki Tools in the Preparation and Support of e-Learning Courses

ERIC Educational Resources Information Center

Jancarik, Antonin; Jancarikova, Katerina

2010-01-01

Wiki tools, which became known mainly thanks to the Wikipedia encyclopedia, represent quite a new phenomenon on the Internet. The work presented here deals with three areas connected to a possible use of wiki tools for the preparation of an e-learning course. To what extent does Wikipedia.com contain terms necessary for scientific lectures at the…

Combination of nano-material enrichment and dead-end filtration for uniform and rapid sample preparation in matrix-assisted laser desorption/ionization mass spectrometry.

PubMed

Wu, Zengnan; Khan, Mashooq; Mao, Sifeng; Lin, Ling; Lin, Jin-Ming

2018-05-01

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a fast analysis tool for the detection of a wide range of analytes. However, heterogeneous distribution of matrix/analyte cocrystal, variation in signal intensity and poor experimental reproducibility at different locations of the same spot means difficulty in quantitative analysis. In this work, carbon nanotubes (CNTs) were employed as adsorbent for analyte cum matrix on a conductive porous membrane as a novel mass target plate. The sample pretreatment step was achieved by enrichment and dead-end filtration and dried by a solid-liquid separation. This approach enables the homogeneous distribution of analyte in the matrix, good shot-to-shot reproducibility in signals and quantitative detection of peptide and protein at different concentrations with correlation coefficient (R 2 ) of 0.9920 and 0.9909, respectively. The simple preparation of sample in a short time, uniform distribution of analyte, easy quantitative detection, and high reproducibility makes this technique useful and may diversify the application of MALDI-MS for quantitative detection of a variety of proteins. Copyright © 2018 Elsevier B.V. All rights reserved.

Using a portable ion mobility spectrometer to screen dietary supplements for sibutramine.

PubMed

Dunn, Jamie D; Gryniewicz-Ruzicka, Connie M; Kauffman, John F; Westenberger, Benjamin J; Buhse, Lucinda F

2011-02-20

In response to recent incidents of undeclared sibutramine, an appetite suppressant found in dietary supplements, we developed a method to detect sibutramine using hand-held ion mobility spectrometers with an analysis time of 15 s. Ion mobility spectrometry is a high-throughput and sensitive technique that has been used for illicit drug, explosive, volatile organic compound and chemical warfare detection. We evaluated a hand-held ion mobility spectrometer as a tool for the analysis of supplement extracts containing sibutramine. The overall instrumental limit of detection of five portable ion mobility spectrometers was 2 ng of sibutramine HCl. When sample extractions containing 30 ng/μl or greater of sibutramine were analyzed, saturation of the ionization chamber of the spectrometer occurred and the instrument required more than three cleaning cycles to remove the drug. Hence, supplement samples suspected of containing sibutramine should be prepared at concentrations of 2-20 ng/μl. To obtain this target concentration range for products containing unknown amounts of sibutramine, we provided a simple sample preparation procedure, allowing the U.S. Food and Drug Administration or other agencies to screen products using the portable ion mobility spectrometer. Published by Elsevier B.V.

Development and Testing of the Church Environment Audit Tool.

PubMed

Kaczynski, Andrew T; Jake-Schoffman, Danielle E; Peters, Nathan A; Dunn, Caroline G; Wilcox, Sara; Forthofer, Melinda

2018-05-01

In this paper, we describe development and reliability testing of a novel tool to evaluate the physical environment of faith-based settings pertaining to opportunities for physical activity (PA) and healthy eating (HE). Tool development was a multistage process including a review of similar tools, stakeholder review, expert feedback, and pilot testing. Final tool sections included indoor opportunities for PA, outdoor opportunities for PA, food preparation equipment, kitchen type, food for purchase, beverages for purchase, and media. Two independent audits were completed at 54 churches. Interrater reliability (IRR) was determined with Kappa and percent agreement. Of 218 items, 102 were assessed for IRR and 116 could not be assessed because they were not present at enough churches. Percent agreement for all 102 items was over 80%. For 42 items, the sample was too homogeneous to assess Kappa. Forty-six of the remaining items had Kappas greater than 0.60 (25 items 0.80-1.00; 21 items 0.60-0.79), indicating substantial to almost perfect agreement. The tool proved reliable and efficient for assessing church environments and identifying potential intervention points. Future work can focus on applications within faith-based partnerships to understand how church environments influence diverse health outcomes.

Preparation and Certification of Two New Bulk Welding Fume Reference Materials for Use in Laboratories Undertaking Analysis of Occupational Hygiene Samples

PubMed Central

Butler, Owen; Musgrove, Darren; Stacey, Peter

2014-01-01

Workers can be exposed to fume, arising from welding activities, which contain toxic metals and metalloids. Occupational hygienists need to assess and ultimately minimize such exposure risks. The monitoring of the concentration of particles in workplace air is one assessment approach whereby fume, from representative welding activities, is sampled onto a filter and returned to a laboratory for analysis. Inductively coupled plasma-atomic emission spectrometry and inductively coupled plasma-mass spectrometry are generally employed as instrumental techniques of choice for the analysis of such filter samples. An inherent difficulty, however, with inductively coupled plasma-based analytical techniques is that they typically require a sample to be presented for analysis in the form of a solution. The efficiency of the required dissolution step relies heavily upon the skill and experience of the analyst involved. A useful tool in assessing the efficacy of this dissolution step would be the availability and subsequent analysis of welding fume reference materials with stated elemental concentrations and matrices that match as closely as possible the matrix composition of welding fume samples submitted to laboratories for analysis. This article describes work undertaken at the Health and Safety Laboratory to prepare and certify two new bulk welding fume reference materials that can be routinely used by analysts to assess the performance of the digestion procedures they employ in their laboratories. PMID:24499055

Preparation and certification of two new bulk welding fume reference materials for use in laboratories undertaking analysis of occupational hygiene samples.

PubMed

Butler, Owen; Musgrove, Darren; Stacey, Peter

2014-01-01

Workers can be exposed to fume, arising from welding activities, which contain toxic metals and metalloids. Occupational hygienists need to assess and ultimately minimize such exposure risks. The monitoring of the concentration of particles in workplace air is one assessment approach whereby fume, from representative welding activities, is sampled onto a filter and returned to a laboratory for analysis. Inductively coupled plasma-atomic emission spectrometry and inductively coupled plasma-mass spectrometry are generally employed as instrumental techniques of choice for the analysis of such filter samples. An inherent difficulty, however, with inductively coupled plasma-based analytical techniques is that they typically require a sample to be presented for analysis in the form of a solution. The efficiency of the required dissolution step relies heavily upon the skill and experience of the analyst involved. A useful tool in assessing the efficacy of this dissolution step would be the availability and subsequent analysis of welding fume reference materials with stated elemental concentrations and matrices that match as closely as possible the matrix composition of welding fume samples submitted to laboratories for analysis. This article describes work undertaken at the Health and Safety Laboratory to prepare and certify two new bulk welding fume reference materials that can be routinely used by analysts to assess the performance of the digestion procedures they employ in their laboratories.

RNA sample preparation applied to gene expression profiling for the horse biological passport.

PubMed

Bailly-Chouriberry, Ludovic; Baudoin, Florent; Cormant, Florence; Glavieux, Yohan; Loup, Benoit; Garcia, Patrice; Popot, Marie-Agnès; Bonnaire, Yves

2017-09-01

The improvement of doping control is an ongoing race. Techniques to fight doping are usually based on the direct detection of drugs or their metabolites by analytical methods such as chromatography hyphenated to mass spectrometry after ad hoc sample preparation. Nowadays, omic methods constitute an attractive development and advances have been achieved particularly by application of molecular biology tools for detection of anabolic androgenic steroids (AAS), erythropoiesis-stimulating agent (ESA), or to control human growth hormone misuses. These interesting results across different animal species have suggested that modification of gene expression offers promising new methods of improving the window of detection of banned substances by targeting their effects on blood cell gene expression. In this context, the present study describes the possibility of using a modified version of the dedicated Human IVD (in vitro Diagnostics) PAXgene® Blood RNA Kit for horse gene expression analysis in blood collected on PAXgene® tubes applied to the horse biological passport. The commercial kit was only approved for human blood samples and has required an optimization of specific technical requirements for equine blood samples. Improvements and recommendations were achieved for sample collection, storage and RNA extraction procedure. Following these developments, RNA yield and quality were demonstrated to be suitable for downstream gene expression analysis by qPCR techniques. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

Quantification of Parvovirus B19 DNA Using COBAS AmpliPrep Automated Sample Preparation and LightCycler Real-Time PCR

PubMed Central

Schorling, Stefan; Schalasta, Gunnar; Enders, Gisela; Zauke, Michael

2004-01-01

The COBAS AmpliPrep instrument (Roche Diagnostics GmbH, D-68305 Mannheim, Germany) automates the entire sample preparation process of nucleic acid isolation from serum or plasma for polymerase chain reaction analysis. We report the analytical performance of the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics) using nucleic acids isolated with the COBAS AmpliPrep instrument. Nucleic acids were extracted using the Total Nucleic Acid Isolation Kit (Roche Diagnostics) and amplified with the LightCycler Parvovirus B19 Quantification Kit. The kit combination processes 72 samples per 8-hour shift. The lower detection limit is 234 IU/ml at a 95% hit-rate, linear range approximately 104-1010 IU/ml, and overall precision 16 to 40%. Relative sensitivity and specificity in routine samples from pregnant women are 100% and 93%, respectively. Identification of a persistent parvovirus B19-infected individual by the polymerase chain reaction among 51 anti-parvovirus B19 IgM-negative samples underlines the importance of additional nucleic acid testing in pregnancy and its superiority to serology in identifying the risk of parvovirus B19 transmission via blood or blood products. Combination of the Total Nucleic Acid Isolation Kit on the COBAS AmpliPrep instrument with the LightCycler Parvovirus B19 Quantification Kit provides a reliable and time-saving tool for sensitive and accurate detection of parvovirus B19 DNA. PMID:14736825

New tool for getting data on the field for paleoclimate and paleoceanography data based on isotope 13C and 18O measurements

NASA Astrophysics Data System (ADS)

Mandic, M.; Stöbener, N.; Smajgl, D.

2017-12-01

For many decades different instrumental methods involving generations of the isotope ratio mass spectrometers with different periphery units for sample preparation, have provided scientifically required high precision, and high throughput of samples for varies application - from geological and hydrological to food and forensic. With this work we introduce automated measurement of δ13C and δ18O from solid carbonate samples, DIC and δ18O of water. We have demonstrated usage of a Thermo Scientific™ Delta Ray™ IRIS with URI Connect on certified reference materials and confirmed the high achievable accuracy and a precision better then <0.1‰ for both δ13C and δ18O, in the laboratory or the field with same precision and throughput of samples. With equilibration method for determination of δ18O in water samples, which we present in this work, achieved repeatability and accuracy are 0.12‰ and 0.68‰ respectively, which fulfill requirements of regulatory methods. The preparation of the samples for carbonate and DIC analysis on the Delta Ray IRIS with URI Connect is similar to the previously mentioned Gas Bench II methods. Samples are put into vials and phosphoric acid is added. The resulting sample-acid chemical reaction releases CO2 gas, which is then introduced into the Delta Ray IRIS via the Variable Volume. Three international standards of carbonate materials (NBS-18, NBS-19 and IAEA-CO-1) were analyzed. NBS-18 and NBS-19 were used as standards for calibration, and IAEA-CO-1 was treated as unknown. For water sample analysis equilibration method with 1% of CO2 in dry air was used. Test measurements and conformation of precision and accuracy of method determination δ18O in water samples were done with three lab standards, namely ANST, OCEAN 2 and HBW. All laboratory standards were previously calibrated with international reference material VSMOW2 and SLAP2 to assure accuracy of the isotopic values. The Principle of Identical Treatment was applied in sample and standard preparation, in measurement procedure, as well as in the evaluation of the results.

doGlycans–Tools for Preparing Carbohydrate Structures for Atomistic Simulations of Glycoproteins, Glycolipids, and Carbohydrate Polymers for GROMACS

PubMed Central

2017-01-01

Carbohydrates constitute a structurally and functionally diverse group of biological molecules and macromolecules. In cells they are involved in, e.g., energy storage, signaling, and cell–cell recognition. All of these phenomena take place in atomistic scales, thus atomistic simulation would be the method of choice to explore how carbohydrates function. However, the progress in the field is limited by the lack of appropriate tools for preparing carbohydrate structures and related topology files for the simulation models. Here we present tools that fill this gap. Applications where the tools discussed in this paper are particularly useful include, among others, the preparation of structures for glycolipids, nanocellulose, and glycans linked to glycoproteins. The molecular structures and simulation files generated by the tools are compatible with GROMACS. PMID:28906114

Measured reflectance suppressed by thin-film interference of crude oil smeared on glass - as on vitrinite in coal or petroliferous rocks

USGS Publications Warehouse

Bostick, Neely

2011-01-01

The tool of measuring "vitrinite reflectance" under a microscope has great value in petroleum exploration and coal utilization, and the reflectance is a simple number, such as 1.4% Ro, with some slight variations depending on technique. Sample collection, preparation and measurement are simple and many sedimentary rocks yield vitrinite. However, the reported number can lead one astray if its origin and quality are not fully understood. I analyze here just one factor, "smear" of crude oil on the polished surface (from the sample), which may reduce reflectance because of thin-film interference. Some other causes of error are listed in an addendum to this note.

Sex Differences in Object Manipulation in Wild Immature Chimpanzees (Pan troglodytes schweinfurthii) and Bonobos (Pan paniscus): Preparation for Tool Use?

PubMed Central

Koops, Kathelijne; Furuichi, Takeshi; Hashimoto, Chie; van Schaik, Carel P.

2015-01-01

Sex differences in immatures predict behavioural differences in adulthood in many mammal species. Because most studies have focused on sex differences in social interactions, little is known about possible sex differences in ‘preparation’ for adult life with regards to tool use skills. We investigated sex and age differences in object manipulation in immature apes. Chimpanzees use a variety of tools across numerous contexts, whereas bonobos use few tools and none in foraging. In both species, a female bias in adult tool use has been reported. We studied object manipulation in immature chimpanzees at Kalinzu (Uganda) and bonobos at Wamba (Democratic Republic of Congo). We tested predictions of the ‘preparation for tool use’ hypothesis. We confirmed that chimpanzees showed higher rates and more diverse types of object manipulation than bonobos. Against expectation, male chimpanzees showed higher object manipulation rates than females, whereas in bonobos no sex difference was found. However, object manipulation by male chimpanzees was play-dominated, whereas manipulation types of female chimpanzees were more diverse (e.g., bite, break, carry). Manipulation by young immatures of both species was similarly dominated by play, but only in chimpanzees did it become more diverse with age. Moreover, in chimpanzees, object types became more tool-like (i.e., sticks) with age, further suggesting preparation for tool use in adulthood. The male bias in object manipulation in immature chimpanzees, along with the late onset of tool-like object manipulation, indicates that not all (early) object manipulation (i.e., object play) in immatures prepares for subsistence tool use. Instead, given the similarity with gender differences in human children, object play may also function in motor skill practice for male-specific behaviours (e.g., dominance displays). In conclusion, even though immature behaviours almost certainly reflect preparation for adult roles, more detailed future work is needed to disentangle possible functions of object manipulation during development. PMID:26444011

Signature-Discovery Approach for Sample Matching of a Nerve-Agent Precursor using Liquid Chromatography–Mass Spectrometry, XCMS, and Chemometrics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fraga, Carlos G.; Clowers, Brian H.; Moore, Ronald J.

2010-05-15

This report demonstrates the use of bioinformatic and chemometric tools on liquid chromatography mass spectrometry (LC-MS) data for the discovery of ultra-trace forensic signatures for sample matching of various stocks of the nerve-agent precursor known as methylphosphonic dichloride (dichlor). The use of the bioinformatic tool known as XCMS was used to comprehensively search and find candidate LC-MS peaks in a known set of dichlor samples. These candidate peaks were down selected to a group of 34 impurity peaks. Hierarchal cluster analysis and factor analysis demonstrated the potential of these 34 impurities peaks for matching samples based on their stock source.more » Only one pair of dichlor stocks was not differentiated from one another. An acceptable chemometric approach for sample matching was determined to be variance scaling and signal averaging of normalized duplicate impurity profiles prior to classification by k-nearest neighbors. Using this approach, a test set of dichlor samples were all correctly matched to their source stock. The sample preparation and LC-MS method permitted the detection of dichlor impurities presumably in the parts-per-trillion (w/w). The detection of a common impurity in all dichlor stocks that were synthesized over a 14-year period and by different manufacturers was an unexpected discovery. Our described signature-discovery approach should be useful in the development of a forensic capability to help in criminal investigations following chemical attacks.« less

KS-Detect – Validation of Solar Thermal PCR for the Diagnosis of Kaposi’s Sarcoma Using Pseudo-Biopsy Samples

PubMed Central

Snodgrass, Ryan; Gardner, Andrea; Jiang, Li; Fu, Cheng; Cesarman, Ethel; Erickson, David

2016-01-01

Resource-limited settings present unique engineering challenges for medical diagnostics. Diagnosis is often needed for those unable to reach central healthcare systems, making portability and independence from traditional energy infrastructure essential device parameters. In 2014, our group presented a microfluidic device that performed a solar-powered variant of the polymerase chain reaction, which we called solar thermal PCR. In this work, we expand on our previous effort by presenting an integrated, portable, solar thermal PCR system targeted towards the diagnosis of Kaposi’s sarcoma. We call this system KS-Detect, and we now report the system’s performance as a diagnostic tool using pseudo-biopsy samples made from varying concentrations of human lymphoma cell lines positive for the KS herpesvirus (KSHV). KS-Detect achieved 83% sensitivity and 70% specificity at high (≥10%) KSHV+ cell concentrations when diagnosing pseudo-biopsy samples by smartphone image. Using histology, we confirm that our prepared pseudo-biopsies contain similar KSHV+ cell concentrations as human biopsies positive for KS. Through our testing of samples derived from human cell lines, we validate KS-Detect as a viable, portable KS diagnostic tool, and we identify critical engineering considerations for future solar-thermal PCR devices. PMID:26799834

Determination of Flavonoids in Wine by High Performance Liquid Chromatography

NASA Astrophysics Data System (ADS)

da Queija, Celeste; Queirós, M. A.; Rodrigues, Ligia M.

2001-02-01

The experiment presented is an application of HPLC to the analysis of flavonoids in wines, designed for students of instrumental methods. It is done in two successive 4-hour laboratory sessions. While the hydrolysis of the wines is in progress, the students prepare the calibration curves with standard solutions of flavonoids and calculate the regression lines and correlation coefficients. During the second session they analyze the hydrolyzed wine samples and calculate the concentrations of the flavonoids using the calibration curves obtained earlier. This laboratory work is very attractive to students because they deal with a common daily product whose components are reported to have preventive and therapeutic effects. Furthermore, students can execute preparative work and apply a more elaborate technique that is nowadays an indispensable tool in instrumental analysis.

Detection of human papillomavirus DNA in urine. A review of the literature.

PubMed

Vorsters, A; Micalessi, I; Bilcke, J; Ieven, M; Bogers, J; Van Damme, P

2012-05-01

The detection of human papillomavirus (HPV) DNA in urine, a specimen easily obtained by a non-invasive self-sampling method, has been the subject of a considerable number of studies. This review provides an overview of 41 published studies; assesses how different methods and settings may contribute to the sometimes contradictory outcomes; and discusses the potential relevance of using urine samples in vaccine trials, disease surveillance, epidemiological studies, and specific settings of cervical cancer screening. Urine sampling, storage conditions, sample preparation, DNA extraction, and DNA amplification may all have an important impact on HPV DNA detection and the form of viral DNA that is detected. Possible trends in HPV DNA prevalence in urine could be inferred from the presence of risk factors or the diagnosis of cervical lesions. HPV DNA detection in urine is feasible and may become a useful tool but necessitates further improvement and standardization.

Histology as a Valid Tool To Differentiate Fresh from Frozen-Thawed Marinated Fish.

PubMed

Meistro, Serena; Pezzolato, Marzia; Muscolino, Daniele; Giarratana, Filippo; Baioni, Elisa; Panebianco, Antonio; Bozzetta, Elena

2016-08-01

European Commission Regulation (EU) 1276/2011 requires that fishery products intended for raw consumption be frozen at -20°C for not less than 24 h or at -35°C for at least 15 h in order to kill viable parasites other than trematodes. But because marinating processes are not always effective in destroying nematode larvae, raw marinated fish preparations should be frozen before consumption. This study evaluated the performance of a standardized histological method to distinguish between fresh and frozen-thawed raw marinated fish. Sixty anchovy (Engraulis encrasicolus) fillets were sampled: 30 were marinated at +4°C for 24 h, and 30 were frozen at -20°C for 24 h before being marinated for 24 h. All 60 samples were fixed in formalin, processed for paraffin embedding, cut, and stained with hematoxylin and eosin. The slide preparations were examined microscopically by three independent histopathologists and classified as frozen-thawed or negative according to standard operating procedure criteria in use at our laboratory. Performance evaluation of the method showed 100% sensitivity (95% confidence interval [CI], 88.4 to 100%) and 100% specificity (95% CI, 88.4 to 100%), and the interrater agreement (Cohen's kappa) was 1 (95% CI, 0.85 to 1). Histology proved a valid and reliable tool to distinguish fresh from frozen-thawed marinated fish. It can be applied to deliver safe raw fishery products to consumers in order to minimize the risk of anisakidosis.

Automation of laboratory testing for infectious diseases using the polymerase chain reaction-- our past, our present, our future.

PubMed

Jungkind, D

2001-01-01

While it is an extremely powerful and versatile assay method, polymerase chain reaction (PCR) can be a labor-intensive process. Since the advent of commercial test kits from Roche and the semi-automated microwell Amplicor system, PCR has become an increasingly useful and widespread clinical tool. However, more widespread acceptance of molecular testing will depend upon automation that allows molecular assays to enter the routine clinical laboratory. The forces driving the need for automated PCR are the requirements for diagnosis and treatment of chronic viral diseases, economic pressures to develop more automated and less expensive test procedures similar to those in the clinical chemistry laboratories, and a shortage in many areas of qualified laboratory personnel trained in the types of manual procedures used in past decades. The automated Roche COBAS AMPLICOR system has automated the amplification and detection process. Specimen preparation remains the most labor-intensive part of the PCR testing process, accounting for the majority of the hands-on-time in most of the assays. A new automated specimen preparation system, the COBAS AmpliPrep, was evaluated. The system automatically releases the target nucleic acid, captures the target with specific oligonucleotide probes, which become attached to magnetic beads via a biotin-streptavidin binding reaction. Once attached to the beads, the target is purified and concentrated automatically. Results of 298 qualitative and 57 quantitative samples representing a wide range of virus concentrations analyzed after the COBAS AmpliPrep and manual specimen preparation methods, showed that there was no significant difference in qualitative or quantitative hepatitis C virus (HCV) assay performance, respectively. The AmpliPrep instrument decreased the time required to prepare serum or plasma samples for HCV PCR to under 1 min per sample. This was a decrease of 76% compared to the manual specimen preparation method. Systems that can analyze more samples with higher throughput and that can answer more questions about the nature of the microbes that we can presently only detect and quantitate will be needed in the future.

Robot based deposition of WC-Co HVOF coatings on HSS cutting tools as a substitution for solid cemented carbide cutting tools

NASA Astrophysics Data System (ADS)

Tillmann, W.; Schaak, C.; Biermann, D.; Aßmuth, R.; Goeke, S.

2017-03-01

Cemented carbide (hard metal) cutting tools are the first choice to machine hard materials or to conduct high performance cutting processes. Main advantages of cemented carbide cutting tools are their high wear resistance (hardness) and good high temperature strength. In contrast, cemented carbide cutting tools are characterized by a low toughness and generate higher production costs, especially due to limited resources. Usually, cemented carbide cutting tools are produced by means of powder metallurgical processes. Compared to conventional manufacturing routes, these processes are more expensive and only a limited number of geometries can be realized. Furthermore, post-processing and preparing the cutting edges in order to achieve high performance tools is often required. In the present paper, an alternative method to substitute solid cemented carbide cutting tools is presented. Cutting tools made of conventional high speed steels (HSS) were coated with thick WC-Co (88/12) layers by means of thermal spraying (HVOF). The challenge is to obtain a dense, homogenous, and near-net-shape coating on the flanks and the cutting edge. For this purpose, different coating strategies were realized using an industrial robot. The coating properties were subsequently investigated. After this initial step, the surfaces of the cutting tools were ground and selected cutting edges were prepared by means of wet abrasive jet machining to achieve a smooth and round micro shape. Machining tests were conducted with these coated, ground and prepared cutting tools. The occurring wear phenomena were analyzed and compared to conventional HSS cutting tools. Overall, the results of the experiments proved that the coating withstands mechanical stresses during machining. In the conducted experiments, the coated cutting tools showed less wear than conventional HSS cutting tools. With respect to the initial wear resistance, additional benefits can be obtained by preparing the cutting edge by means of wet abrasive jet machining.

FT-Raman and chemometric tools for rapid determination of quality parameters in milk powder: Classification of samples for the presence of lactose and fraud detection by addition of maltodextrin.

PubMed

Rodrigues Júnior, Paulo Henrique; de Sá Oliveira, Kamila; de Almeida, Carlos Eduardo Rocha; De Oliveira, Luiz Fernando Cappa; Stephani, Rodrigo; Pinto, Michele da Silva; de Carvalho, Antônio Fernandes; Perrone, Ítalo Tuler

2016-04-01

FT-Raman spectroscopy has been explored as a quick screening method to evaluate the presence of lactose and identify milk powder samples adulterated with maltodextrin (2.5-50% w/w). Raman measurements can easily differentiate samples of milk powder, without the need for sample preparation, while traditional quality control methods, including high performance liquid chromatography, are cumbersome and slow. FT-Raman spectra were obtained from samples of whole lactose and low-lactose milk powder, both without and with addition of maltodextrin. Differences were observed between the spectra involved in identifying samples with low lactose content, as well as adulterated samples. Exploratory data analysis using Raman spectroscopy and multivariate analysis was also developed to classify samples with PCA and PLS-DA. The PLS-DA models obtained allowed to correctly classify all samples. These results demonstrate the utility of FT-Raman spectroscopy in combination with chemometrics to infer about the quality of milk powder. Copyright © 2015 Elsevier Ltd. All rights reserved.

Implementing self sustained quality control procedures in a clinical laboratory.

PubMed

Khatri, Roshan; K C, Sanjay; Shrestha, Prabodh; Sinha, J N

2013-01-01

Quality control is an essential component in every clinical laboratory which maintains the excellence of laboratory standards, supplementing to proper disease diagnosis, patient care and resulting in overall strengthening of health care system. Numerous quality control schemes are available, with combinations of procedures, most of which are tedious, time consuming and can be "too technical" whereas commercially available quality control materials can be expensive especially for laboratories in developing nations like Nepal. Here, we present a procedure performed at our centre with self prepared control serum and use of simple statistical tools for quality assurance. The pooled serum was prepared as per guidelines for preparation of stabilized liquid quality control serum from human sera. Internal Quality Assessment was performed on this sample, on a daily basis which included measurement of 12 routine biochemical parameters. The results were plotted on Levey-Jennings charts and analysed with quality control rules, for a period of one month. The mean levels of biochemical analytes in self prepared control serum were within normal physiological range. This serum was evaluated every day along with patients' samples. The results obtained were plotted on control charts and analysed using common quality control rules to identify possible systematic and random errors. Immediate mitigation measures were taken and the dispatch of erroneous reports was avoided. In this study we try to highlight on a simple internal quality control procedure which can be performed by laboratories, with minimum technology, expenditure, and expertise and improve reliability and validity of the test reports.

Optimization of the β-Elimination/Michael Addition Chemistry on Reversed-Phase Supports for Mass Spectrometry Analysis of O-Linked Protein Modifications

PubMed Central

Nika, Heinz; Nieves, Edward; Hawke, David H.; Angeletti, Ruth Hogue

2013-01-01

We previously adapted the β-elimination/Michael addition chemistry to solid-phase derivatization on reversed-phase supports, and demonstrated the utility of this reaction format to prepare phosphoseryl peptides in unfractionated protein digests for mass spectrometric identification and facile phosphorylation-site determination. Here, we have expanded the use of this technique to β-N-acetylglucosamine peptides, modified at serine/threonine, phosphothreonyl peptides, and phosphoseryl/phosphothreonyl peptides, followed in sequence by proline. The consecutive β-elimination with Michael addition was adapted to optimize the solid-phase reaction conditions for throughput and completeness of derivatization. The analyte remained intact during derivatization and was recovered efficiently from the silica-based, reversed-phase support with minimal sample loss. The general use of the solid-phase approach for enzymatic dephosphorylation was demonstrated with phosphoseryl and phosphothreonyl peptides and was used as an orthogonal method to confirm the identity of phosphopeptides in proteolytic mixtures. The solid-phase approach proved highly suitable to prepare substrates from low-level amounts of protein digests for phosphorylation-site determination by chemical-targeted proteolysis. The solid-phase protocol provides for a simple, robust, and efficient tool to prepare samples for phosphopeptide identification in MALDI mass maps of unfractionated protein digests, using standard equipment available in most biological laboratories. The use of a solid-phase analytical platform is expected to be readily expanded to prepare digest from O-glycosylated- and O-sulfonated proteins for mass spectrometry-based structural characterization. PMID:23997661

Effect of preparation methods and doping on the structural and tunable emissions of CdS

NASA Astrophysics Data System (ADS)

Mohamed, Mohamed Bakr; Abdel-Kader, M. H.; Alhazime, Ali A.; Almarashi, Jamal Q. M.

2018-03-01

Fe, Mn and Mg doped CdS samples were prepared by thermolysis method in air and under flow of nitrogen. Structural, compositional and optical properties of the prepared samples were investigated using x-ray powder diffraction (XRD), scanning electron microscope (SEM/EDS mapping), Fourier transform infrared red (FTIR), UV-vis absorption and photoluminescence (PL) spectroscopes. Rietveld refinement of x-ray data showed that all the undoped and doped CdS samples prepared in air and under flow of nitrogen have both cubic and hexagonal structures. The percentages of hexagonal and cubic phases for all prepared samples were determined. The crystallite size increased for CdS prepared under flow of N2 compared with the sample prepared in air. The energy gap of all the samples was calculated using UV data. The intensity of PL emission changed according to the method of preparation and the kind of doping elements. PL emission revealed a blue shift for CdS prepared in air compared with CdS prepared under flow of nitrogen; also all doped samples showed a red shift of PL spectra compared with undoped samples. Undoped and doped CdS with Fe and Mg samples emitted violet and blue sub-spectra. Mn doped CdS prepared in air revealed violet, blue and yellow sub-spectra, while the sample prepared under flow of N2 emitted violet, blue and green sub-spectra.

Thermo-Mechanical Effect on Poly Crystalline Boron Nitride Tool Life During Friction Stir Welding (Dwell Period)

NASA Astrophysics Data System (ADS)

Almoussawi, M.; Smith, A. J.

2018-05-01

Poly Crystalline Boron Nitride (PCBN) tool wear during the friction stir welding of high melting alloys is an obstacle to commercialize the process. This work simulates the friction stir welding process and tool wear during the plunge/dwell period of 14.8 mm EH46 thick plate steel. The Computational Fluid Dynamic (CFD) model was used for simulation and the wear of the tool is estimated from temperatures and shear stress profile on the tool surface. Two sets of tool rotational speeds were applied including 120 and 200 RPM. Seven plunge/dwell samples were prepared using PCBN FSW tool, six thermocouples were also embedded around each plunge/dwell case in order to record the temperatures during the welding process. Infinite focus microscopy technique was used to create macrographs for each case. The CFD result has been shown that a shear layer around the tool shoulder and probe-side denoted as thermo-mechanical affected zone (TMAZ) was formed and its size increase with tool rotational speed increase. Maximum peak temperature was also found to increase with tool rotational speed increase. PCBN tool wear under shoulder was found to increase with tool rotational speed increase as a result of tool's binder softening after reaching to a peak temperature exceeds 1250 °C. Tool wear also found to increase at probe-side bottom as a result of high shear stress associated with the decrease in the tool rotational speed. The amount of BN particles revealed by SEM in the TMAZ were compared with the CFD model.

Adhesive bonded structural repair. II - Surface preparation procedures, tools, equipment and facilities

NASA Astrophysics Data System (ADS)

Wegman, Raymond F.; Tullos, Thomas R.

1993-10-01

A development status report is presented on the surface preparation procedures, tools, equipment, and facilities used in adhesively-bonded repair of aerospace and similar high-performance structures. These methods extend to both metallic and polymeric surfaces. Attention is given to the phos-anodize containment system, paint removal processes, tools for cutting composite prepreg and fabric materials, autoclaves, curing ovens, vacuum bagging, and controlled atmospheres.

Apparatus for preparing cornea material for tabbed (sutureless) transplantation

DOEpatents

Collins, Joseph Patrick

1997-01-01

A tool and a method for preparing a donor material used in sutureless corneal transplants uses a first cutting portion to prepare a donor blank having tabbed portions extending outwardly radially. A second cutting portion is used to cut the central portion of the blank. The tool is used as a guide member for the second cutting portion. In one embodiment the tool has slits laterally defined therethrough which allow the tabbed portions of the donor material to be thinned to a desired thickness using a scalpel. In an another embodiment the second cutting portion is a round trephine which is used to simultaneously trim each of the tabbed portions.

Exploring Operational Test and Evaluation of Unmanned Aircraft Systems: A Qualitative Case Study

NASA Astrophysics Data System (ADS)

Saliceti, Jose A.

The purpose of this qualitative case study was to explore and identify strategies that may potentially remedy operational test and evaluation procedures used to evaluate Unmanned Aircraft Systems (UAS) technology. The sample for analysis consisted of organizations testing and evaluating UASs (e.g., U.S. Air Force, U.S. Navy, U.S. Army, U.S. Marine Corps, U.S. Coast Guard, and Customs Border Protection). A purposeful sampling technique was used to select 15 subject matter experts in the field of operational test and evaluation of UASs. A questionnaire was provided to participants to construct a descriptive and robust research. Analysis of responses revealed themes related to each research question. Findings revealed operational testers utilized requirements documents to extrapolate measures for testing UAS technology and develop critical operational issues. The requirements documents were (a) developed without the contribution of stakeholders and operational testers, (b) developed with vague or unrealistic measures, and (c) developed without a systematic method to derive requirements from mission tasks. Four approaches are recommended to develop testable operational requirements and assist operational testers: (a) use a mission task analysis tool to derive requirements for mission essential tasks for the system, (b) exercise collaboration among stakeholders and testers to ensure testable operational requirements based on mission tasks, (c) ensure testable measures are used in requirements documents, and (d) create a repository list of critical operational issues by mission areas. The preparation of operational test and evaluation processes for UAS technology is not uniform across testers. The processes in place are not standardized, thus test plan preparation and reporting are different among participants. A standard method to prepare and report UAS technology should be used when preparing and reporting on UAS technology. Using a systematic process, such as mission-based test design, resonated among participants as an analytical method to link UAS mission tasks and measures of performance to the capabilities of the system under test when developing operational test plans. Further research should examine system engineering designs for system requirements traceability matrix of mission tasks and subtasks while using an analysis tool that adequately evaluates UASs with an acceptable level of confidence in the results.

Developing a tool for the preparation of GMP audit of pharmaceutical contract manufacturer.

PubMed

Linna, Anu; Korhonen, Mirka; Mannermaa, Jukka-Pekka; Airaksinen, Marja; Juppo, Anne Mari

2008-06-01

Outsourcing is rapidly growing in the pharmaceutical industry. When the manufacturing activities are outsourced, control of the product's quality has to be maintained. One way to confirm contract manufacturer's GMP (Good Manufacturing Practice) compliance is auditing. Audits can be supported for instance by using GMP questionnaires. The objective of this study was to develop a tool for the audit preparation of pharmaceutical contract manufacturers and to validate its contents by using Delphi method. At this phase of the study the tool was developed for non-sterile finished product contract manufacturers. A modified Delphi method was used with expert panel consisting of 14 experts from pharmaceutical industry, authorities and university. The content validity of the developed tool was assessed by a Delphi questionnaire round. The response rate in Delphi questionnaire round was 86%. The tool consisted of 103 quality items, from which 90 (87%) achieved the pre-defined agreement rate level (75%). Thirteen quality items which did not achieve the pre-defined agreement rate were excluded from the tool. The expert panel suggested only minor changes to the tool. The results show that the content validity of the developed audit preparation tool was good.

Artificial Urine for Teaching Urinalysis Concepts and Diagnosis of Urinary Tract Infection in the Medical Microbiology Laboratory.

PubMed

Khan, Latifa B; Read, Hannah M; Ritchie, Stephen R; Proft, Thomas

2017-01-01

Dipstick urinalysis is an informative, quick, cost-effective and non-invasive diagnostic tool that is useful in clinical practice for the diagnosis of urinary tract infections (UTIs), kidney diseases, and diabetes. We used dipstick urinalysis as a hands-on microbiology laboratory exercise to reinforce student learning about UTIs with a particular focus on cystitis, which is a common bacterial infection. To avoid exposure to potentially contaminated human urine samples, we prepared artificial urine using easily acquired and affordable ingredients, which allowed less-experienced students to perform urinalysis without the risk of exposure to pathogenic organisms and ensured reliable availability of the urine samples. This practical class taught medical students how to use urinalysis data in conjunction with medical history to diagnose diseases from urine samples and to determine a treatment plan for clinical scenarios.

Attempt to assess the infiltration of enamel made with experimental preparation using a scanning electron microscope.

PubMed

Skucha-Nowak, Małgorzata

2015-01-01

The resin infiltration technique, a minimally invasive method, involves the saturation, strengthening, and stabilization of demineralized enamel by a mixture of polymer resins without the need to use rotary tools or the risk of losing healthy tooth structures. To design and synthesize an experimental infiltrant with potential bacteriostatic properties.To compare the depth of infiltration of the designed experimental preparation with the infiltrant available in the market using a scanning electron microscope. Composition of the experimental infiltrant was established after analysis of 1H NMR spectra of the commercially available compounds that can penetrate pores of demineralized enamel. As the infiltrant should have bacteriostatic features by definition, an addition of 1% of monomer containing metronidazole was made. Thirty extracted human teeth were soaked in an acidic solution, which was to provide appropriate conditions for demineralization of enamel. Afterward, each tooth was divided along the coronal-root axis into two zones. One zone had experimental preparation applied to it (the test group), while the other had commercially available Icon (the control group). The teeth were dissected along the long axis and described above underwent initial observation with use of a Hitachi S-4200 scanning electron microscope. It was found that all samples contained only oxygen and carbon, regardless of the concentration of additions introduced into them. The occurrence of carbon is partially because it is a component of the preparation in question and partially because of sputtering of the sample with it. Hydrogen is also a component of the preparation, as a result of its phase composition; however, it cannot be detected by the EDS method. SEM, in combination with X-ray microanalysis, does not allow one to explicitly assess the depth of penetration of infiltration preparations into enamel.In order to assess the depth of penetration of infiltration preparations with use of X-ray microanalysis, it is recommended to introduce a contrast agent that is approved for use in dental materials, such as ytterbium III fluoride.

The Science of and Advanced Technology for Cost-Effective Manufacture of High Precision Engineering Products. Volume 4. Thermal Effects on the Accuracy of Numerically Controlled Machine Tools.

DTIC Science & Technology

1985-10-01

83K0385 FINAL REPORT D Vol. 4 00 THERMAL EFFECTS ON THE ACCURACY OF LD NUME" 1ICALLY CONTROLLED MACHINE TOOLS PREPARED BY I Raghunath Venugopal and M...OF NUMERICALLY CONTROLLED MACHINE TOOLS 12 PERSONAL AJ’HOR(S) Venunorial, Raghunath and M. M. Barash 13a TYPE OF REPORT 13b TIME COVERED 14 DATE OF...TOOLS Prepared by Raghunath Venugopal and M. M. Barash Accesion For Unannounced 0 Justification ........................................... October 1085

Effect of Processing on Silk-Based Biomaterials: Reproducibility and Biocompatibility

PubMed Central

Wray, Lindsay S.; Hu, Xiao; Gallego, Jabier; Georgakoudi, Irene; Omenetto, Fiorenzo G.; Schmidt, Daniel; Kaplan, David L.

2012-01-01

Silk fibroin has been successfully used as a biomaterial for tissue regeneration. In order to prepare silk fibroin biomaterials for human implantation a series of processing steps are required to purify the protein. Degumming to remove inflammatory sericin is a crucial step related to biocompatibility and variability in the material. Detailed characterization of silk fibroin degumming is reported. The degumming conditions significantly affected cell viability on the silk fibroin material and the ability to form three-dimensional porous scaffolds from the silk fibroin, but did not affect macrophage activation or β-sheet content in the materials formed. Methods are also provided to determine the content of residual sericin in silk fibroin solutions and to assess changes in silk fibroin molecular weight. Amino acid composition analysis was used to detect sericin residuals in silk solutions with a detection limit between 1.0% and 10% wt/wt, while fluorescence spectroscopy was used to reproducibly distinguish between silk samples with different molecular weights. Both methods are simple and require minimal sample volume, providing useful quality control tools for silk fibroin preparation processes. PMID:21695778

Liquid-based cytology and cell block immunocytochemistry in veterinary medicine: comparison with standard cytology for the evaluation of canine lymphoid samples.

PubMed

Fernandes, N C C A; Guerra, J M; Réssio, R A; Wasques, D G; Etlinger-Colonelli, D; Lorente, S; Nogueira, E; Dagli, M L Z

2016-08-01

Liquid-based Cytology (LBC) consists of immediate wet cell fixation with automated slide preparation. We applied LBC, cell block (CB) and immunocytochemistry to diagnose canine lymphoma and compare results with conventional cytology. Samples from enlarged lymph nodes of 18 dogs were collected and fixed in preservative solution for automated slide preparation (LBC), CB inclusion and immunophenotyping. Two CB techniques were tested: fixed sediment method (FSM) and agar method (AM). Anti-CD79a, anti-Pax5, anti-CD3 and anti-Ki67 were used in immunocytochemistry. LBC smears showed better nuclear and nucleolar definition, without cell superposition, but presented smaller cell size and worse cytoplasmic definition. FSM showed consistent cellular groups and were employed for immunocytochemistry, while AM CBs presented sparse groups of lymphocytes, with compromised analysis. Anti-Pax-5 allowed B-cell identification, both in reactive and neoplastic lymph nodes. Our preliminary report suggests that LBC and FSM together may be promising tools to improve lymphoma diagnosis through fine-needle aspiration. © 2015 John Wiley & Sons Ltd.

Preparing and Analyzing Iced Airfoils

NASA Technical Reports Server (NTRS)

Vickerman, Mary B.; Baez, Marivell; Braun, Donald C.; Cotton, Barbara J.; Choo, Yung K.; Coroneos, Rula M.; Pennline, James A.; Hackenberg, Anthony W.; Schilling, Herbert W.; Slater, John W.;

Comparison of conventional twist drill protocol and piezosurgery for implant insertion: an ex vivo study on different bone types.

PubMed

Sagheb, Keyvan; Kumar, Vinay V; Azaripour, Adriano; Walter, Christian; Al-Nawas, Bilal; Kämmerer, Peer W

2017-02-01

The aim of this ex vivo study was to compare implant insertion procedures using piezosurgery and conventional drilling in different qualities of bone. Implant bed preparation time, generated heat, and primary implant stability were analyzed. Fresh ex vivo porcine bone block samples (cancellous, mixed, and cortical bone) were obtained. The bone quality was quantified by ultrasound transmission velocity (UTV). Each bone sample received three implants of the same diameter using each of the techniques of piezosurgery and conventional twist drills. Time for preparation was taken and the temperature while performing the osteotomy was measured using infrared spectroscopy. The primary implant stability after osteotomy was measured using resonance frequency analysis (RFA) and extrusion torque (ET). ANOVA with post hoc Tukey test was carried out to compare the values for the three different groups. The UTV values strongly correlated with the density of the bone samples. There was a significant increase in time (threefold, P < 0.05 [302 s vs. 122 s in cortical bone]) but no difference in the temperature for the piezo group (~37°C in cortical bone). Regardless of the osteotomy technique, there was a statistically significant increase in RFA and ET values in implants inserted in cancellous bone (RFA: piezo 77, drill 76; ET: piezo 22, drill 21), mixed bone (RFA: piezo 85, drill 86; ET: piezo 105, drill 61), and cortical bone (RFA: piezo 90, drill 87; ET piezo 184, drill 79) samples, respectively (P < 0.05). In between the different osteotomy groups, there was no difference in the RFA values but significant higher ET values in mixed/cortical bone samples in favor for the piezosurgery group. Piezosurgery and conventional implant bed drilling procedure do have similar mechanical outcomes regarding primary stability with high RFA values, but the preparation does need more time for piezosurgery group, so that piezosurgery might be a valuable tool in only very specific cases for implant bed preparation. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

7 CFR 27.21 - Preparation of samples of cotton.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 2 2012-01-01 2012-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this section...

7 CFR 27.21 - Preparation of samples of cotton.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this section...

7 CFR 27.21 - Preparation of samples of cotton.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this section...

Evaluation of visible and near-infrared spectroscopy as a tool for assessing fiber fineness during mechanical preparation of dew-retted flax.

PubMed

Sharma, H S S; Reinard, N

2004-12-01

Flax fiber must be mechanically prepared to improve fineness and homogeneity of the sliver before chemical processing and wet-spinning. The changes in fiber characteristics are monitored by an airflow method, which is labor intensive and requires 90 minutes to process one sample. This investigation was carried out to develop robust visible and near-infrared calibrations that can be used as a rapid tool for quality assessment of input fibers and changes in fineness at the doubling (blending), first, second, third, and fourth drawing frames, and at the roving stage. The partial least squares (PLS) and principal component regression (PCR) methods were employed to generate models from different segments of the spectra (400-1100, 1100-1700, 1100-2498, 1700-2498, and 400-2498 nm) and a calibration set consisting of 462 samples obtained from the six processing stages. The calibrations were successfully validated with an independent set of 97 samples, and standard errors of prediction of 2.32 and 2.62 dtex were achieved with the best PLS (400-2498 nm) and PCR (1100-2498 nm) models, respectively. An optimized PLS model of the visible-near-infrared (vis-NIR) spectra explained 97% of the variation (R(2) = 0.97) in the sample set with a standard error of calibration (SEC) of 2.45 dtex and a standard error of cross-validation (SECV) of 2.51 dtex R(2) = 0.96). The mean error of the reference airflow method was 1.56 dtex, which is more accurate than the NIR calibration. The improvement in fiber fineness of the validation set obtained from the six production lines was predicted with an error range of -6.47 to +7.19 dtex for input fibers, -1.44 to +5.77 dtex for blended fibers at the doubling, and -4.72 to +3.59 dtex at the drawing frame stages. This level of precision is adequate for wet-spinners to monitor fiber fineness of input fibers and during the preparation of fibers. The advantage of visNIR spectroscopy is the potential capability of the technique to assess fineness and other important quality characteristics of a fiber sample simultaneously in less than 30 minutes; the disadvantages are the expensive instrumentation and the expertise required for operating the instrument compared to the reference method. These factors need to be considered by the industry before installing an off-line NIR system for predicting quality parameters of input materials and changes in fiber characteristics during mechanical processing.

Visualizing Morphological Changes of Abscission Zone Cells in Arabidopsis by Scanning Electron Microscope.

PubMed

Shi, Chun-Lin; Butenko, Melinka A

2018-01-01

Scanning electron microscope (SEM) is a type of electron microscope which produces detailed images of surface structures. It has been widely used in plants and animals to study cellular structures. Here, we describe a detailed protocol to prepare samples of floral abscission zones (AZs) for SEM, as well as further image analysis. We show that it is a powerful tool to detect morphologic changes at the cellular level during the course of abscission in wild-type plants and to establish the details of phenotypic alteration in abscission mutants.

STS-87 Day 14 Highlights

NASA Technical Reports Server (NTRS)

1997-01-01

On this fourteenth day of the STS-87 mission, the flight crew, Cmdr. Kevin R. Kregel, Pilot Steven W. Lindsey, Mission Specialists Winston E. Scott, Kalpana Chawla, and Takao Doi, and Payload Specialist Leonid K. Kadenyuk focus on completion of hands-on sample processing in the microgravity glovebox facility. They also prepare the spacesuits and tools that will be used for the EVA by Scott and Doi. The crew take time out from their schedule to discuss the mission with reporters from the U.S., Japan and the Ukraine during the traditional in-flight news conference.

Optical Fabrication and Measurement AXAF and CIRS

NASA Technical Reports Server (NTRS)

Engelhaupt, Darell

1997-01-01

This paper presents a final report on Optical Fabrication and Measurement AXAF (Advanced X-Ray Astrophysics Facility) and CIRS (Composite Infrared Spectrometer) from July 12, 1994 to August 16, 1996.. This paper includes specific tasks to be performed. The tasks are as follows: 1) Preparation and Characterization of Zerodur Glass Samples; 2) Develop and Fabricate AXAF and CIRS Metrology Tooling; 3) Update AXAF Technical Data Base; and 4) Perform Fabrication Related Metrology Tasks for CIRS. This paper also includes final activities from the July, 1996 report to August 1996.

Assessing and calibrating the ATR-FTIR approach as a carbonate rock characterization tool

NASA Astrophysics Data System (ADS)

Henry, Delano G.; Watson, Jonathan S.; John, Cédric M.

2017-01-01

ATR-FTIR (attenuated total reflectance Fourier transform infrared) spectroscopy can be used as a rapid and economical tool for qualitative identification of carbonates, calcium sulphates, oxides and silicates, as well as quantitatively estimating the concentration of minerals. Over 200 powdered samples with known concentrations of two, three, four and five phase mixtures were made, then a suite of calibration curves were derived that can be used to quantify the minerals. The calibration curves in this study have an R2 that range from 0.93-0.99, a RMSE (root mean square error) of 1-5 wt.% and a maximum error of 3-10 wt.%. The calibration curves were used on 35 geological samples that have previously been studied using XRD (X-ray diffraction). The identification of the minerals using ATR-FTIR is comparable with XRD and the quantitative results have a RMSD (root mean square deviation) of 14% and 12% for calcite and dolomite respectively when compared to XRD results. ATR-FTIR is a rapid technique (identification and quantification takes < 5 min) that involves virtually no cost if the machine is available. It is a common tool in most analytical laboratories, but it also has the potential to be deployed on a rig for real-time data acquisition of the mineralogy of cores and rock chips at the surface as there is no need for special sample preparation, rapid data collection and easy analysis.

A tool for the estimation of the distribution of landslide area in R

NASA Astrophysics Data System (ADS)

Rossi, M.; Cardinali, M.; Fiorucci, F.; Marchesini, I.; Mondini, A. C.; Santangelo, M.; Ghosh, S.; Riguer, D. E. L.; Lahousse, T.; Chang, K. T.; Guzzetti, F.

2012-04-01

We have developed a tool in R (the free software environment for statistical computing, http://www.r-project.org/) to estimate the probability density and the frequency density of landslide area. The tool implements parametric and non-parametric approaches to the estimation of the probability density and the frequency density of landslide area, including: (i) Histogram Density Estimation (HDE), (ii) Kernel Density Estimation (KDE), and (iii) Maximum Likelihood Estimation (MLE). The tool is available as a standard Open Geospatial Consortium (OGC) Web Processing Service (WPS), and is accessible through the web using different GIS software clients. We tested the tool to compare Double Pareto and Inverse Gamma models for the probability density of landslide area in different geological, morphological and climatological settings, and to compare landslides shown in inventory maps prepared using different mapping techniques, including (i) field mapping, (ii) visual interpretation of monoscopic and stereoscopic aerial photographs, (iii) visual interpretation of monoscopic and stereoscopic VHR satellite images and (iv) semi-automatic detection and mapping from VHR satellite images. Results show that both models are applicable in different geomorphological settings. In most cases the two models provided very similar results. Non-parametric estimation methods (i.e., HDE and KDE) provided reasonable results for all the tested landslide datasets. For some of the datasets, MLE failed to provide a result, for convergence problems. The two tested models (Double Pareto and Inverse Gamma) resulted in very similar results for large and very large datasets (> 150 samples). Differences in the modeling results were observed for small datasets affected by systematic biases. A distinct rollover was observed in all analyzed landslide datasets, except for a few datasets obtained from landslide inventories prepared through field mapping or by semi-automatic mapping from VHR satellite imagery. The tool can also be used to evaluate the probability density and the frequency density of landslide volume.

Targeted Observation of ELL Instruction as a Tool in the Preparation of School Leaders

ERIC Educational Resources Information Center

Baecher, Laura; Knoll, Marcia; Patti, Janet

2016-01-01

Preparing school administrators to promote effective instruction of English language learners (ELLs) is an important dimension of today's educational leadership programs, requiring innovative program activities. This study explores school leadership candidates' use of an observation tool targeted to ELL instruction that incorporated guided video…

In vitro synthesis and characterisation of three fenoterol sulfoconjugates detected in fenoterol post-administration urine samples.

PubMed

Orlovius, A K; Guddat, S; Gütschow, M; Thevis, M; Schänzer, W

2013-11-01

Fenoterol, a fast-acting β2-adrenergic agonist, is used in the therapy of obstructive pulmonary diseases and for the inhibition of premature labour obstetrics. Doping control for β2-agonists, which are prohibited in sports by the World Anti-Doping Agency, is commonly performed by liquid chromatography/mass spectrometry after hydrolysis of phase II metabolites. The continuing development of analytical procedures has led to direct injection of urine samples without sample preparation becoming a viable tool. For the detection of substances without sample preparation, including hydrolysis, detailed information of the phase II metabolism of the substances is essential. In this study, human S9 fractions of different tissues and two recombinant sulfotransferases were investigated for their potential to form fenoterol sulfoconjugates, which were characterised in detail. Two mono-sulfoconjugates and one bis-sulfoconjugate were synthesised and their structures confirmed by liquid chromatography–high-resolution/high-accuracy mass spectrometry. All of the metabolites were identified as esterified phenolic compounds. Excretion studies with orally and inhalatively administered fenoterol proved the occurrence of the sulfoconjugates in vivo. Inhalatively administered fenoterol resulted in the detection of the two monosulfoconjugates in low amounts in urine due to the lower inhalation dose of fenoterol compared to the oral dose. After oral uptake of fenoterol, the two mono-sulfoconjugates and a fenoterol bis-sulfoconjugate were detected in urine. This is the first report of the bis-sulfoconjugate.

Label-free virus detection using silicon photonic microring resonators.

PubMed

McClellan, Melinda S; Domier, Leslie L; Bailey, Ryan C

2012-01-15

Viruses represent a continual threat to humans through a number of mechanisms, which include disease, bioterrorism, and destruction of both plant and animal food resources. Many contemporary techniques used for the detection of viruses and viral infections suffer from limitations such as the need for extensive sample preparation or the lengthy window between infection and measurable immune response, for serological methods. In order to develop a method that is fast, cost-effective, and features reduced sample preparation compared to many other virus detection methods, we report the application of silicon photonic microring resonators for the direct, label-free detection of intact viruses in both purified samples as well as in a complex, real-world analytical matrix. As a model system, we demonstrate the quantitative detection of Bean pod mottle virus, a pathogen of great agricultural importance, with a limit of detection of 10 ng/mL. By simply grinding a small amount of leaf sample in buffer with a mortar and pestle, infected leaves can be identified over a healthy control with a total analysis time of less than 45 min. Given the inherent scalability and multiplexing capability of the semiconductor-based technology, we feel that silicon photonic microring resonators are well-positioned as a promising analytical tool for a number of viral detection applications. Copyright © 2011 Elsevier B.V. All rights reserved.

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

PubMed Central

Maddi, Balakrishna; Panisko, Ellen; Albrecht, Karl; Howe, Daniel

2016-01-01

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in complex mixtures. It is often used to analyze gasoline, jet fuel, diesel, bio-diesel and the organic fraction of bio-crude/bio-oil. In most of those analyses, the first dimension of separation is non-polar, followed by a polar separation. The aqueous fractions of bio-crude and other aqueous samples from biofuels production have been examined with similar column combinations. However, sample preparation techniques such as derivatization, solvent extraction, and solid-phase extraction were necessaryprior to analysis. In this study, aqueous fractions obtained from the hydrothermal liquefaction of algae were characterized by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry without prior sample preparation techniques using a polar separation in the first dimension followed by a non-polar separation in the second. Two-dimensional plots from this analysis were compared with those obtained from the more traditional column configuration. Results from qualitative characterization of the aqueous fractions of algal bio-crude are discussed in detail. The advantages of using a polar separation followed by a non-polar separation for characterization of organics in aqueous samples by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry are highlighted. PMID:27022829

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry.

PubMed

Maddi, Balakrishna; Panisko, Ellen; Albrecht, Karl; Howe, Daniel

2016-03-06

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in complex mixtures. It is often used to analyze gasoline, jet fuel, diesel, bio-diesel and the organic fraction of bio-crude/bio-oil. In most of those analyses, the first dimension of separation is non-polar, followed by a polar separation. The aqueous fractions of bio-crude and other aqueous samples from biofuels production have been examined with similar column combinations. However, sample preparation techniques such as derivatization, solvent extraction, and solid-phase extraction were necessary prior to analysis. In this study, aqueous fractions obtained from the hydrothermal liquefaction of algae were characterized by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry without prior sample preparation techniques using a polar separation in the first dimension followed by a non-polar separation in the second. Two-dimensional plots from this analysis were compared with those obtained from the more traditional column configuration. Results from qualitative characterization of the aqueous fractions of algal bio-crude are discussed in detail. The advantages of using a polar separation followed by a non-polar separation for characterization of organics in aqueous samples by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry are highlighted.

Preparation of longitudinal sections of hair samples for the analysis of cocaine by MALDI-MS/MS and TOF-SIMS imaging.

PubMed

Flinders, Bryn; Cuypers, Eva; Zeijlemaker, Hans; Tytgat, Jan; Heeren, Ron M A

2015-10-01

Matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) for the analysis of intact hair is a powerful tool for the detection of drugs of abuse in toxicology and forensic applications. Here we present a quick, easy, and reproducible method of preparing longitudinal sections of single hairs. This method improves the accessibility of chemicals embedded in the hair matrix for molecular imaging with mass spectrometry. The images obtained from a single, sectioned hair sample show molecular distributions in the exposed medulla, cortex, and a portion of the cuticle observed as a narrow layer surrounding the cortex. Using MALDI-MS/MS imaging, the distribution of cocaine was observed throughout five longitudinally sectioned drug-user hair samples. The images showed the distribution of the product ion at m/z 182, derived from the precursor ion of cocaine at m/z 304. MetA-SIMS images of longitudinally sectioned hair samples showed a more detailed distribution of cocaine at m/z 304, benzoylecgonine the major metabolite of cocaine at m/z 290 and other drugs such as methadone which was observed at m/z 310. Chronological information of drug intake can be obtained more sensitively. The chronological detail is in hours rather than months, which is of great interest in clinical as well as forensic applications. Copyright © 2015 John Wiley & Sons, Ltd.

Surface-enhanced Raman scattering spectroscopy characterization and identification of foodborne bacteria

NASA Astrophysics Data System (ADS)

Liu, Yongliang; Chen, Yud-Ren; Nou, Xiangwu; Chao, Kaunglin

2007-09-01

Rapid and routine identification of foodborne bacteria are considerably important, because of bio- / agro- terrorism threats, public health concerns, and economic loss. Conventional, PCR, and immunoassay methods for the detection of bacteria are generally time-consuming, chemical reagent necessary and multi-step procedures. Fast microbial detection requires minimal sample preparation, permits the routine analysis of large numbers of samples with negligible reagent costs, and is easy to operate. Therefore, we have developed silver colloidal nanoparticle based surface-enhanced Raman scattering (SERS) spectroscopy as a potential tool for the rapid and routine detection of E. coli and L. monocytogenes. This study presents the further results of our examination on S. typhimonium, one of the most commonly outbreak bacteria, for the characteristic bands and subsequent identification.

Nano-plasmonic exosome diagnostics

PubMed Central

Im, Hyungsoon; Shao, Huilin; Weissleder, Ralph; Castro, Cesar M.; Lee, Hakho

2015-01-01

Exosomes have emerged as a promising biomarker. These vesicles abound in biofluids and harbor molecular constituents from their parent cells, thereby offering a minimally-invasive avenue for molecular analyses. Despite such clinical potential, routine exosomal analysis, particularly the protein assay, remains challenging, due to requirements for large sample volumes and extensive processing. We have been developing miniaturized systems to facilitate clinical exosome studies. These systems can be categorized into two components: microfluidics for sample preparation and analytical tools for protein analyses. In this report, we review a new assay platform, nano-plasmonic exosome (nPLEX), in which sensing is based on surface plasmon resonance to achieve label-free exosome detection. Looking forward, we also discuss some potential challenges and improvements in exosome studies. PMID:25936957

Graphic products used in the evaluation of traditional and emerging remote sensing technologies for the detection of fugitive contamination at selected superfund hazardous waste sites

USGS Publications Warehouse

Slonecker, E. Terrence; Fisher, Gary B.

2011-01-01

This report presents the overhead imagery and field sampling results used to prepare U.S. Geological Survey Open-File Report 2011-1050, 'Evaluation of Traditional and Emerging Remote Sensing Technologies for the Detection of Fugitive Contamination at Selected Superfund Hazardous Waste Sites'. These graphic products were used in the evaluation of remote sensing technology in postclosure monitoring of hazardous waste sites and represent an ongoing research effort. Soil sampling results presented here were accomplished with field portable x-ray fluoresence (XRF) technology and are used as screening tools only representing the current conditions of metals and other contaminants at selected Superfund hazardous waste sites.

Implementation of a digital preparation validation tool in dental skills laboratory training.

PubMed

Kozarovska, A; Larsson, C

2018-05-01

To describe the implementation of a digital tool for preparation validation and evaluate it as an aid in students' self-assessment. Students at the final semester of skills laboratory training were asked to use a digital preparation validation tool (PVT) when performing two different tasks; preparation of crowns for teeth 11 and 21. The students were divided into two groups. Group A self-assessed and scanned all three attempts at 21 ("prep-and-scan"). Group B self-assessed all attempts chose the best one and scanned it ("best-of-three"). The situation was reversed for 11. The students assessed five parameters of the preparation and marked them as approved (A) or failed (F). These marks were compared with the information from the PVT. The students also completed a questionnaire. Each question was rated from 1 to 5. Teachers' opinions were collected at staff meetings throughout the project. Most students in the "prep-and-scan" groups showed an increase in agreement between their self-assessment and the information from the PVT, whereas students in the "best-of-three" groups showed lower levels of agreement. All students rated the PVT positively. Most strongly agreed that the tool was helpful in developing skills (mean 4.15), easy to use (mean 4.23) and that it added benefits in comparison to existing assessment tools (mean 4.05). They did not however, fully agree that the tool is time efficient (mean 2.55), and they did not consider it a substitute for verbal teacher feedback. Teachers' feedback suggested advantages of the tool in the form of ease of use, visual aid and increasing interest and motivation during skills laboratory training however, they did not notice a reduction in need of verbal feedback. Within the limitations of the study, our conclusion is that a digital PVT may be a valuable adjunct to other assessment tools in skills laboratory training. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Enhanced bacterial affinity of PVDF membrane: its application as improved sea water sampling tool for environmental monitoring.

PubMed

Kumar, Sweta Binod; Sharnagat, Preeti; Manna, Paramita; Bhattacharya, Amit; Haldar, Soumya

2017-02-01

Isolation of diversified bacteria from seawater is a major challenge in the field of environmental microbiology. In the present study, an attempt has been made to select specific membrane with improved property of attaching diversified bacteria. Initially, different concentrations (15, 18, and 20% W/W) of polysulfone (PSF) were used to check their affinity for the attachment of selected gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria. Among these, 20% W/W PSF showed maximum attachment. Therefore, membrane prepared with other materials such as polyvinylidene fluoride (PVDF) and polyether sulfone (PES) were used with the same concentration (20% W/W) to check their improved bacterial attachment property. Comparative study of bacterial attachment on three different membranes revealed that PVDF possessed the highest affinity towards both the groups of bacteria. This property was confirmed by different analytical methods viz. contact angle, atomic force microscopy, zeta potential, and flux study and further validated with seawater samples collected from seven sites of western coast and Lakshadweep island of India, using Biolog EcoPlate™. All the samples showed that bacterial richness and diversity was high in PVDF membrane in comparison to surrounding seawater samples. Interestingly, affinity for more diversified bacteria was reported to be higher in water sample with less turbidity and low bacteria load. This finding can facilitate the development of PVDF (20% W/W) membrane as a simple, cheap, and less labor intensive environmental sampling tool for the isolation of diversified bacteria from seawater sample wih different physiochemical properties. Graphical abstract ᅟ.

A Gravity-Driven Microfluidic Particle Sorting Device with Hydrodynamic Separation Amplification

PubMed Central

Huh, Dongeun; Bahng, Joong Hwan; Ling, Yibo; Wei, Hsien-Hung; Kripfgans, Oliver D.; Fowlkes, J. Brian; Grotberg, James B.; Takayama, Shuichi

2008-01-01

This paper describes a simple microfluidic sorting system that can perform size-profiling and continuous mass-dependent separation of particles through combined use of gravity (1g) and hydrodynamic flows capable of rapidly amplifying sedimentation-based separation between particles. Operation of the device relies on two microfluidic transport processes: i) initial hydrodynamic focusing of particles in a microchannel oriented parallel to gravity, ii) subsequent sample separation where positional difference between particles with different mass generated by sedimentation is further amplified by hydrodynamic flows whose streamlines gradually widen out due to the geometry of a widening microchannel oriented perpendicular to gravity. The microfluidic sorting device was fabricated in poly(dimethylsiloxane) (PDMS), and hydrodynamic flows in microchannels were driven by gravity without using external pumps. We conducted theoretical and experimental studies on fluid dynamic characteristics of laminar flows in widening microchannels and hydrodynamic amplification of particle separation. Direct trajectory monitoring, collection, and post-analysis of separated particles were performed using polystyrene microbeads with different sizes to demonstrate rapid (< 1 min) and high-purity (> 99.9 %) separation. Finally, we demonstrated biomedical applications of our system by isolating small-sized (diameter < 6 μm) perfluorocarbon liquid droplets from polydisperse droplet emulsions, which is crucial in preparing contrast agents for safe, reliable ultrasound medical imaging, tracers for magnetic resonance imaging, or transpulmonary droplets used in ultrasound-based occlusion therapy for cancer treatment. Our method enables straightforward, rapid real-time size-monitoring and continuous separation of particles in simple stand-alone microfabricated devices without the need for bulky and complex external power sources. We believe that this system will provide a useful tool o separate colloids and particles for various analytical and preparative applications, and may hold 3 potential for separation of cells or development of diagnostic tools requiring point-of-care sample preparation or testing. PMID:17297936

Tools for T-RFLP data analysis using Excel.

PubMed

Fredriksson, Nils Johan; Hermansson, Malte; Wilén, Britt-Marie

2014-11-08

Terminal restriction fragment length polymorphism (T-RFLP) analysis is a DNA-fingerprinting method that can be used for comparisons of the microbial community composition in a large number of samples. There is no consensus on how T-RFLP data should be treated and analyzed before comparisons between samples are made, and several different approaches have been proposed in the literature. The analysis of T-RFLP data can be cumbersome and time-consuming, and for large datasets manual data analysis is not feasible. The currently available tools for automated T-RFLP analysis, although valuable, offer little flexibility, and few, if any, options regarding what methods to use. To enable comparisons and combinations of different data treatment methods an analysis template and an extensive collection of macros for T-RFLP data analysis using Microsoft Excel were developed. The Tools for T-RFLP data analysis template provides procedures for the analysis of large T-RFLP datasets including application of a noise baseline threshold and setting of the analysis range, normalization and alignment of replicate profiles, generation of consensus profiles, normalization and alignment of consensus profiles and final analysis of the samples including calculation of association coefficients and diversity index. The procedures are designed so that in all analysis steps, from the initial preparation of the data to the final comparison of the samples, there are various different options available. The parameters regarding analysis range, noise baseline, T-RF alignment and generation of consensus profiles are all given by the user and several different methods are available for normalization of the T-RF profiles. In each step, the user can also choose to base the calculations on either peak height data or peak area data. The Tools for T-RFLP data analysis template enables an objective and flexible analysis of large T-RFLP datasets in a widely used spreadsheet application.

Apparatus for preparing cornea material for tabbed (sutureless) transplantation

DOEpatents

Collins, J.P.

1997-07-22

A tool and a method for preparing a donor material used in sutureless corneal transplants uses a first cutting portion to prepare a donor blank having tabbed portions extending outwardly radially. A second cutting portion is used to cut the central portion of the blank. The tool is used as a guide member for the second cutting portion. In one embodiment the tool has slits laterally defined therethrough which allow the tabbed portions of the donor material to be thinned to a desired thickness using a scalpel. In an another embodiment the second cutting portion is a round trephine which is used to simultaneously trim each of the tabbed portions. 26 figs.

Nutritional deficiency during colonoscopy preparation: the forgotten iatrogeny.

PubMed

Nunes, Gonçalo; Barata, Ana Teresa; Santos, Carla Adriana; Patita, Marta; Fonseca, Jorge

2018-05-01

bowel preparation for colonoscopy induces a semi-fasting state, with a potential negative impact on fragile patients. The present study aims to quantify nutritional deficiency during colonoscopy preparation. this was an observational and cross-sectional study. A convenience sample was obtained that included adults that underwent colonoscopy after bowel preparation with Klean-Prep® according to the center protocol. Anthropometric evaluation was performed and nutritional deficiency was calculated via the quantification of energy and protein intake during the 48 hours prior to the examination which was compared with the individuals' needs. The association between nutritional deficiency with the quality of bowel preparation, age and status (hospitalized/ambulatory) was evaluated. the study included 131 patients aged 21-91 years (mean 63.6 ± 13.2 years); 73 cases were male. Malnutrition reached 67.2% using specific anthropometric tools. A median preparation quality of six points was found when the Boston Bowel Preparation Scale was considered. The mean intake 48 hours prior to the procedure was 1,795 kcal and 100 g of protein. A daily energy intake of less than 50% of the individual needs was observed in 88 patients and less than 25% in 29 cases. The mean energy and protein deficiency were 59% (p < 0.01) and 45% (p < 0.01), and there was no correlation with preparation quality (p > 0.05). Nutritional defiency is similar in hospitalized and ambulatory patients (p > 0.05), but higher in older individuals (p = 0.04). nutritional deficiency during colonoscopy preparation was significant, more so in older patients, and there was no correlation with the quality of bowel preparation. We conclude that bowel preparation regimens should be reformulated with an improved nutritional intake and the inclusion of nutritional supplements without residues.

Transfer of ochratoxin A from raw black tea to tea infusions prepared according to the Turkish tradition.

PubMed

Toman, Jakub; Malir, Frantisek; Ostry, Vladimir; Kilic, Mehmet Akif; Roubal, Tomas; Grosse, Yann; Pfohl-Leszkowicz, Annie

2018-01-01

Ochratoxin A (OTA) is a natural contaminant of food including tea with multiple toxic effects, which poses a threat to human health. In terms of lifestyle, the Turkish population is a frequent visitor of tearooms, and the traditional Turkish tea preparation is one of the most popular ways of preparing tea infusion. The aim of this study was to investigate OTA transfer from raw black tea to the tea infusion prepared according to the Turkish tradition. A high-performance liquid chromatography method with a limit of quantification of 0.35 ng g -1 was used for OTA determination. The OTA amount in raw black teas from Turkey ranged from ≤0.35 ng g -1 up to 56.7 ng g -1 . An homogenised sample of black tea naturally contaminated with 55.0 ng g -1 was used to prepare infusions. The OTA transfer from the black tea to the infusion was found to be 41.5% ± 7%. These data are important for the realisation of a 'Total Diet study' (TDS). The TDS can be a complementary tool to estimate the population dietary exposure to OTA across the entire diet by analysing main foods prepared 'as consumed' (tea infusions) and not 'as purchased' (raw tea). © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Three-phase molecularly imprinted sol-gel based hollow fiber liquid-phase microextraction combined with liquid chromatography-tandem mass spectrometry for enrichment and selective determination of a tentative lung cancer biomarker.

PubMed

Moein, Mohammad Mahdi; Javanbakht, Mehran; Karimi, Mohammad; Akbari-Adergani, Behrouz; Abdel-Rehim, Mohamed

2015-07-15

In the present study, the modification of a polysulfone hollow fiber membrane with in situ molecularly imprinted sol-gel process (as a novel and one-step method) was prepared and investigated. 3-(propylmethacrylate)trimethoxysilane (3PMTMOS) as an inorganic precursor was used for preparation of molecularly imprinted sol-gel. The modified molecularly imprinted sol-gel hollow fiber membrane (MSHM) was used for the liquid-phase microextraction (LPME) of hippuric acid (HA) in human plasma and urine samples. MSHM as a selective, robust, and durable tool was used for at least 50 extractions without significant decrease in the extraction efficiency. The non-molecularly imprinted sol-gel hollow fiber membrane (NSHM) as blank hollow fiber membrane was prepared by the same process, only without HA. To achieve the best condition, influential parameters on the extraction efficiency were thoroughly investigated. The capability of this robust, green, and simple method for extraction of HA was successfully accomplished with LC/MS/MS. The limits of detection (LOD) and quantification (LOQ) in human plasma and urine samples were 0.3 and 1.0nmolL(-1), respectively. The standard calibration curves were obtained within the concentration range 1-2000nmolL(-1) for HA in human plasma and urine. The coefficients of determination (r(2)) were ≥0.998. The obtained data exhibited recoveries were higher than 89% for the extraction of HA in human plasma and urine samples. Copyright © 2015 Elsevier B.V. All rights reserved.

Metabolomic analysis using porcine skin: a pilot study of analytical techniques.

PubMed

Wu, Julie; Fiehn, Oliver; Armstrong, April W

2014-06-15

Metabolic byproducts serve as indicators of the chemical processes and can provide valuable information on pathogenesis by measuring the amplified output. Standardized techniques for metabolome extraction of skin samples serve as a critical foundation to this field but have not been developed. We sought to determine the optimal cell lysage techniques for skin sample preparation and to compare GC-TOF-MS and UHPLC-QTOF-MS for metabolomic analysis. Using porcine skin samples, we pulverized the skin via various combinations of mechanical techniques for cell lysage. After extraction, the samples were subjected to GC-TOF-MS and/or UHPLC-QTOF-MS. Signal intensities from GC-TOF-MS analysis showed that ultrasonication (2.7x107) was most effective for cell lysage when compared to mortar-and-pestle (2.6x107), ball mill followed by ultrasonication (1.6x107), mortar-and-pestle followed by ultrasonication (1.4x107), and homogenization (trial 1: 8.4x106; trial 2: 1.6x107). Due to the similar signal intensities, ultrasonication and mortar-and-pestle were applied to additional samples and subjected to GC-TOF-MS and UHPLC-QTOF-MS. Ultrasonication yielded greater signal intensities than mortar-and-pestle for 92% of detected metabolites following GC-TOF-MS and for 68% of detected metabolites following UHPLC-QTOF-MS. Overall, ultrasonication is the preferred method for efficient cell lysage of skin tissue for both metabolomic platforms. With standardized sample preparation, metabolomic analysis of skin can serve as a powerful tool in elucidating underlying biological processes in dermatological conditions.

7A projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals.

PubMed

Norville, Julie E; Kelly, Deborah F; Knight, Thomas F; Belcher, Angela M; Walz, Thomas

2007-12-01

Two-dimensional crystallization on lipid monolayers is a versatile tool to obtain structural information of proteins by electron microscopy. An inherent problem with this approach is to prepare samples in a way that preserves the crystalline order of the protein array and produces specimens that are sufficiently flat for high-resolution data collection at high tilt angles. As a test specimen to optimize the preparation of lipid monolayer crystals for electron microscopy imaging, we used the S-layer protein sbpA, a protein with potential for designing arrays of both biological and inorganic materials with engineered properties for a variety of nanotechnology applications. Sugar embedding is currently considered the best method to prepare two-dimensional crystals of membrane proteins reconstituted into lipid bilayers. We found that using a loop to transfer lipid monolayer crystals to an electron microscopy grid followed by embedding in trehalose and quick-freezing in liquid ethane also yielded the highest resolution images for sbpA lipid monolayer crystals. Using images of specimens prepared in this way we could calculate a projection map of sbpA at 7A resolution, one of the highest resolution projection structures obtained with lipid monolayer crystals to date.

Developing Technology Teachers: Questioning the Industrial Tool Use Model

ERIC Educational Resources Information Center

Hansen, John W.; Lovedahl, Gerald G.

2004-01-01

In this article, the author questions the role of technology teacher preparation programs that are based on an "industrial tool use" model to develop technology education teachers. It is the position of the authors that the manner by which technology education teachers are prepared may need revision and that technology teacher educators…

Educate at Penn State: Preparing Beginning Teachers with Powerful Digital Tools

ERIC Educational Resources Information Center

Murray, Orrin T.; Zembal-Saul, Carla

2008-01-01

University based teacher education programs are slowly beginning to catch up to other professional programs that use modern digital tools to prepare students to enter professional fields. This discussion looks at how one teacher education program reached the conclusion that students and faculty would use notebook computers. Frequently referred to…

Improved Dielectric Properties via Mechano-Chemical Activation in Ba0.80Pb0.20TiO3 Ceramics

NASA Astrophysics Data System (ADS)

Kumar, Parveen; Rani, Renu; Singh, Sangeeta; Juneja, J. K.; Prakash, Chandra; Raina, K. K.

2011-12-01

The present report is about the preparation and dielectric properties of commonly used Ba0.80Pb0.20TiO3 (BPT) ferroelectric ceramic via Mechano-Chemical Activation (MCA). Results were compared by the BPT sample prepared by conventional solid state method. The BPT sample prepared via MCA technique was found to have decreased tetragonality, dielectric constant value (ɛRT = 450 and ɛmax = 6170) approximately double the value for sample prepared by conventional method (ɛRT = 260 and ɛmax = 3275). Also, the sample prepared by MCA was found to be less frequency dependent. Thus, the BPT sample prepared via MCA is more suitable for capacitor applications requiring lesser frequency dependency than the conventionally prepared BPT sample.

Raman spectroscopy as a tool to understand Kerogen production potential

NASA Astrophysics Data System (ADS)

Khatibi, S.; Ostadhassan, M.; Mohammed, R. A.; Alexeyev, A.

2017-12-01

A lot attention has given to unconventional reservoirs specifically oil shale in North America during the last decades. Understanding Kerogen properties in terms of maturity and production potential are crucial for unconventional reservoir. Since, the amount of hydrocarbon generation is a function of kerogen type and content in the formation, and the magnitude and duration in which heat and pressure were applied. This study presents a non-destructive and fast method to determine Kerogen properties in terms of Rock-Eval parameters by means of Raman Spectroscopy. Samples were gathered from upper and lower Bakken formation, with different maturities at different depth. Raman spectroscopy as a powerful nondestructive analytical tool for molecular reconstruction was employed to find Raman spectra of different samples. In the next step, Rock-Eval was performed for each sample and different measurements were made. Then in an original approach, correlation between Rock-Eval parameters with Raman Spectroscopy results was established to fully understand how kerogen productivity potentials can be reflected on the Raman response. Results showed, maturity related parameters (RO, Tmax), S1 (already generated oil in the rock), S2 (potential hydrocarbon) and OSI (oil saturation index as indication of potential oil flow zones) can be correlated to band separation, D band intensity, G band intensity and G/D intensity, respectively. Proposed method provide a fast nondestructive method to evaluate Kerogen quality even at field without any special sample preparation.

Environmental swabs as a tool in norovirus outbreak investigation, including outbreaks on cruise ships.

PubMed

Boxman, Ingeborg L A; Dijkman, Remco; te Loeke, Nathalie A J M; Hägele, Geke; Tilburg, Jeroen J H C; Vennema, Harry; Koopmans, Marion

2009-01-01

In this study, we investigated whether environmental swabs can be used to demonstrate the presence of norovirus in outbreak settings. First, a procedure was set up based on viral RNA extraction using guanidium isothiocyanate buffer and binding of nucleic acids to silica. Subsequently, environmental swabs were taken at 23 Dutch restaurants and four cruise ships involved in outbreaks of gastroenteritis. Outbreaks were selected based on clinical symptoms consistent with viral gastroenteritis and time between consumption of suspected food and onset of clinical symptoms (>12 h). Norovirus RNA was demonstrated by real-time reverse transcriptase PCR in 51 of 86 (59%) clinical specimens from 12 of 14 outbreaks (86%), in 13 of 90 (14%) food specimens from 4 of 18 outbreaks (22%), and in 48 of 119 (40%) swab specimens taken from 14 of 27 outbreaks (52%). Positive swab samples agreed with positive clinical samples in seven outbreaks, showing identical sequences. Furthermore, norovirus was detected on swabs taken from kitchen and bathroom surfaces in five outbreaks in which no clinical samples were collected and two outbreaks with negative fecal samples. The detection rate was highest for outbreaks associated with catered meals and lowest for restaurant-associated outbreaks. The use of environmental swabs may be a useful tool in addition to testing of food and clinical specimens, particularlywhen viral RNA is detected on surfaces used for food preparation.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Herrmann, A M; Ritz, K; Nunan, N

Soils are structurally heterogeneous across a wide range of spatio-temporal scales. Consequently, external environmental conditions do not have a uniform effect throughout the soil, resulting in a large diversity of micro-habitats. It has been suggested that soil function can be studied without explicit consideration of such fine detail, but recent research has indicated that the micro-scale distribution of organisms may be of importance for a mechanistic understanding of many soil functions. Due to a lack of techniques with adequate sensitivity for data collection at appropriate scales, the question 'How important are various soil processes acting at different scales for ecologicalmore » function?' is challenging to answer. The nano-scale secondary ion mass spectrometer (NanoSIMS) represents the latest generation of ion microprobes which link high-resolution microscopy with isotopic analysis. The main advantage of NanoSIMS over other secondary ion mass spectrometers is the ability to operate at high mass resolution, whilst maintaining both excellent signal transmission and spatial resolution ({approx}50 nm). NanoSIMS has been used previously in studies focusing on presolar materials from meteorites, in material science, biology, geology and mineralogy. Recently, the potential of NanoSIMS as a new tool in the study of biophysical interfaces in soils has been demonstrated. This paper describes the principles of NanoSIMS and discusses the potential of this tool to contribute to the field of biogeochemistry and soil ecology. Practical considerations (sample size and preparation, simultaneous collection of isotopes, mass resolution, isobaric interference and quantification of the isotopes of interest) are discussed. Adequate sample preparation avoiding biases in the interpretation of NanoSIMS data due to artifacts and identification of regions-of interest are of most concerns in using NanoSIMS as a new tool in biogeochemistry and soil ecology. Finally, we review the areas of research most likely to benefit from the high resolving power attainable with this new approach.« less

Stable-Carbon Isotope Ratios for Sourcing the Nerve-Agent Precursor Methylphosphonic Dichloride and Its Products

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moran, James J.; Fraga, Carlos G.; Nims, Megan K.

The ability to connect a chemical threat agent to a specific batch of a synthetic precursor can provide a fingerprint to contribute to effective forensic investigations. Stable isotope analysis can leverage intrinsic, natural isotopic variability within the molecules of a threat agent to unlock embedded chemical fingerprints in the material. Methylphosphonic dichloride (DC) is a chemical precursor to the nerve agent sarin. DC is converted to methylphosphonic difluoride (DF) as part of the sarin synthesis process. We used a suite of commercially available DC stocks to both evaluate the potential for δ 13C analysis to be used as a fingerprintingmore » tool in sarin-related investigations and to develop sample preparation techniques (using chemical hydrolysis) that can simplify isotopic analysis of DC and its synthetic products. We demonstrate that natural isotopic variability in DC results in at least three distinct, isotope-resolved clusters within the thirteen stocks we analyzed. Isotopic variability in the carbon feedstock (i.e., methanol) used for DC synthesis is likely inherited by the DC samples we measured. Here, we demonstrate that the hydrolysis of DC and DF to methylphosphonic acid (MPA) can be used as a preparative step for isotopic analysis because the reaction does not impart a significant isotopic fractionation. MPA is more chemically stable, less toxic, and easier to handle than DC or DF. Further, the hydrolysis method we demonstrated can be applied to a suite of other precursors or to sarin itself, thereby providing a potentially valuable forensic tool.« less

Stable-Carbon Isotope Ratios for Sourcing the Nerve-Agent Precursor Methylphosphonic Dichloride and Its Products

DOE PAGES

Moran, James J.; Fraga, Carlos G.; Nims, Megan K.

2018-04-01

The ability to connect a chemical threat agent to a specific batch of a synthetic precursor can provide a fingerprint to contribute to effective forensic investigations. Stable isotope analysis can leverage intrinsic, natural isotopic variability within the molecules of a threat agent to unlock embedded chemical fingerprints in the material. Methylphosphonic dichloride (DC) is a chemical precursor to the nerve agent sarin. DC is converted to methylphosphonic difluoride (DF) as part of the sarin synthesis process. We used a suite of commercially available DC stocks to both evaluate the potential for δ 13C analysis to be used as a fingerprintingmore » tool in sarin-related investigations and to develop sample preparation techniques (using chemical hydrolysis) that can simplify isotopic analysis of DC and its synthetic products. We demonstrate that natural isotopic variability in DC results in at least three distinct, isotope-resolved clusters within the thirteen stocks we analyzed. Isotopic variability in the carbon feedstock (i.e., methanol) used for DC synthesis is likely inherited by the DC samples we measured. Here, we demonstrate that the hydrolysis of DC and DF to methylphosphonic acid (MPA) can be used as a preparative step for isotopic analysis because the reaction does not impart a significant isotopic fractionation. MPA is more chemically stable, less toxic, and easier to handle than DC or DF. Further, the hydrolysis method we demonstrated can be applied to a suite of other precursors or to sarin itself, thereby providing a potentially valuable forensic tool.« less

Stable-carbon isotope ratios for sourcing the nerve-agent precursor methylphosphonic dichloride and its products.

PubMed

Moran, James J; Fraga, Carlos G; Nims, Megan K

2018-08-15

The ability to connect a chemical threat agent to a specific batch of a synthetic precursor can provide a fingerprint to contribute to effective forensic investigations. Stable isotope analysis can leverage intrinsic, natural isotopic variability within the molecules of a threat agent to unlock embedded chemical fingerprints in the material. Methylphosphonic dichloride (DC) is a chemical precursor to the nerve agent sarin. DC is converted to methylphosphonic difluoride (DF) as part of the sarin synthesis process. We used a suite of commercially available DC stocks to both evaluate the potential for δ 13 C analysis to be used as a fingerprinting tool in sarin-related investigations and to develop sample preparation techniques (using chemical hydrolysis) that can simplify isotopic analysis of DC and its synthetic products. We demonstrate that natural isotopic variability in DC results in at least three distinct, isotope-resolved clusters within the thirteen stocks we analyzed. Isotopic variability in the carbon feedstock (i.e., methanol) used for DC synthesis is likely inherited by the DC samples we measured. We demonstrate that the hydrolysis of DC and DF to methylphosphonic acid (MPA) can be used as a preparative step for isotopic analysis because the reaction does not impart a significant isotopic fractionation. MPA is more chemically stable, less toxic, and easier to handle than DC or DF. Further, the hydrolysis method we demonstrated can be applied to a suite of other precursors or to sarin itself, thereby providing a potentially valuable forensic tool. Copyright © 2018. Published by Elsevier B.V.

Changes of the elemental distributions in marine diatoms as a reporter of sample preparation artefacts. A nuclear microscopy application

NASA Astrophysics Data System (ADS)

Godinho, R. M.; Cabrita, M. T.; Alves, L. C.; Pinheiro, T.

2015-04-01

Studies of the elemental composition of whole marine diatoms cells have high interest as they constitute a direct measurement of environmental changes, and allow anticipating consequences of anthropogenic alterations to organisms, ecosystems and global marine geochemical cycles. Nuclear microscopy is a powerful tool allowing direct measurement of whole cells giving qualitative imaging of distribution, and quantitative determination of intracellular concentration. Major obstacles to the analysis of marine microalgae are high medium salinity and the recurrent presence of extracellular exudates produced by algae to maintain colonies in natural media and in vitro. The objective of this paper was to optimize the methodology of sample preparation of marine unicellular algae for elemental analysis with nuclear microscopy, allowing further studies on cellular response to metals. Primary cultures of Coscinodiscus wailesii maintained in vitro were used to optimize protocols for elemental analysis with nuclear microscopy techniques. Adequate cell preparation procedures to isolate the cells from media components and exudates were established. The use of chemical agents proved to be inappropriate for elemental determination and for intracellular morphological analysis. The assessment of morphology and elemental partitioning in cell compartments obtained with nuclear microscopy techniques enabled to infer their function in natural environment and imbalances in exposure condition. Exposure to metal affected C. wailesii morphology and internal elemental distribution.

Next-generation confirmatory disease diagnostics

NASA Astrophysics Data System (ADS)

Lin, Robert; Gerver, Rachel; Karns, Kelly; Apori, Akwasi A.; Denisin, Aleksandra K.; Herr, Amy E.

2014-06-01

Microfluidic tools are advancing capabilities in screening diagnostics for use in near-patient settings. Here, we review three case studies to illustrate the flexibility and analytical power offered by microanalytical tools. We first overview a near-patient tool for detection of protein markers found in cerebrospinal fluid (CSF), as a means to identify the presence of cerebrospinal fluid in nasal mucous - an indication that CSF is leaking into the nasal cavity. Microfluidic design allowed integration of several up-stream preparatory steps and rapid, specific completion of the human CSF protein assay. Second, we overview a tear fluid based assay for lactoferrin, a protein produced in the lacrimal gland, then secreted into tear fluid. Tear Lf is a putative biomarker for primary SS. A critical contribution of this and related work being measurement of Lf, even in light of well-known and significant matrix interactions and losses during the tear fluid collection and preparation. Lastly, we review a microfluidic barcode platform that enables rapid measurement of multiple infectious disease biomarkers in human sera. The assay presents a new approach to multiplexed biomarker detection, yet in a simple straight microchannel - thus providing a streamlined, simplified microanalytical platform, as is relevant to robust operation in diagnostic settings. We view microfluidic design and analytical chemistry as the basis for emerging, sophisticated assays that will advance not just screening diagnostic technology, but confirmatory assays, sample preparation and handling, and thus introduction and utilization of new biomarkers and assay formats.

The Recent Developments in Sample Preparation for Mass Spectrometry-Based Metabolomics.

PubMed

Gong, Zhi-Gang; Hu, Jing; Wu, Xi; Xu, Yong-Jiang

2017-07-04

Metabolomics is a critical member in systems biology. Although great progress has been achieved in metabolomics, there are still some problems in sample preparation, data processing and data interpretation. In this review, we intend to explore the roles, challenges and trends in sample preparation for mass spectrometry- (MS-) based metabolomics. The newly emerged sample preparation methods were also critically examined, including laser microdissection, in vivo sampling, dried blood spot, microwave, ultrasound and enzyme-assisted extraction, as well as microextraction techniques. Finally, we provide some conclusions and perspectives for sample preparation in MS-based metabolomics.

From near-infrared and Raman to surface-enhanced Raman spectroscopy: progress, limitations and perspectives in bioanalysis.

PubMed

Dumont, Elodie; De Bleye, Charlotte; Sacré, Pierre-Yves; Netchacovitch, Lauranne; Hubert, Philippe; Ziemons, Eric

2016-05-01

Over recent decades, spreading environmental concern entailed the expansion of green chemistry analytical tools. Vibrational spectroscopy, belonging to this class of analytical tool, is particularly interesting taking into account its numerous advantages such as fast data acquisition and no sample preparation. In this context, near-infrared, Raman and mainly surface-enhanced Raman spectroscopy (SERS) have thus gained interest in many fields including bioanalysis. The two former techniques only ensure the analysis of concentrated compounds in simple matrices, whereas the emergence of SERS improved the performances of vibrational spectroscopy to very sensitive and selective analyses. Complex SERS substrates were also developed enabling biomarker measurements, paving the way for SERS immunoassays. Therefore, in this paper, the strengths and weaknesses of these techniques will be highlighted with a focus on recent progress.

Simultaneous enantioselective quantification of fluoxetine and norfluoxetine in human milk by direct sample injection using 2-dimensional liquid chromatography-tandem mass spectrometry.

PubMed

Alvim, Joel; Lopes, Bianca Rebelo; Cass, Quezia Bezerra

2016-06-17

A two-dimensional liquid chromatography system coupled to triple quadrupole tandem mass spectrometer (2D LC-MS/MS) was employed for the simultaneously quantification of fluoxetine (FLX) and norfluoxetine (NFLX) enantiomers in human milk by direct injection of samples. A restricted access media of bovine serum albumin octadecyl column (RAM-BSAC18) was used in the first dimension for the milk proteins depletion, while an antibiotic-based chiral column was used in the second dimension. The results herein described show good selectivity, extraction efficiency, accuracy, and precision with limits of quantification in the order of 7.5ngmL(-1)for the FLX enantiomers and 10.0ngmL(-1) for NFLX enantiomers. Furthermore, it represents a practical tool in terms of sustainability for the sample preparation of such a difficult matrix. Copyright © 2016 Elsevier B.V. All rights reserved.

Qualitative carbonyl profile in coffee beans through GDME-HPLC-DAD-MS/MS for coffee preliminary characterization.

PubMed

Cordeiro, Liliana; Valente, Inês M; Santos, João Rodrigo; Rodrigues, José A

2018-05-01

In this work, an analytical methodology for volatile carbonyl compounds characterization in green and roasted coffee beans was developed. The methodology relied on a recent and simple sample preparation technique, gas diffusion microextraction for extraction of the samples' volatiles, followed HPLC-DAD-MS/MS analysis. The experimental conditions in terms of extraction temperature and extraction time were studied. A profile for carbonyl compounds was obtained for both arabica and robusta coffee species (green and roasted samples). Twenty-seven carbonyl compounds were identified and further discussed, in light of reported literature, with different coffee characteristics: coffee ageing, organoleptic impact, presence of defective beans, authenticity, human's health implication, post-harvest coffee processing and roasting. The applied methodology showed to be a powerful analytical tool to be used for coffee characterization as it measures marker compounds of different coffee characteristics. Copyright © 2018 Elsevier Ltd. All rights reserved.

Medical Applications of Laser Induced Breakdown Spectroscopy

NASA Astrophysics Data System (ADS)

Pathak, A. K.; Rai, N. K.; Singh, Ankita; Rai, A. K.; Rai, Pradeep K.; Rai, Pramod K.

2014-11-01

Sedentary lifestyle of human beings has resulted in various diseases and in turn we require a potential tool that can be used to address various issues related to human health. Laser Induced Breakdown Spectroscopy (LIBS) is one such potential optical analytical tool that has become quite popular because of its distinctive features that include applicability to any type/phase of samples with almost no sample preparation. Several reports are available that discusses the capabilities of LIBS, suitable for various applications in different branches of science which cannot be addressed by traditional analytical methods but only few reports are available for the medical applications of LIBS. In the present work, LIBS has been implemented to understand the role of various elements in the formation of gallstones (formed under the empyema and mucocele state of gallbladder) samples along with patient history that were collected from Purvancal region of Uttar Pradesh, India. The occurrence statistics of gallstones under the present study reveal higher occurrence of gallstones in female patients. The gallstone occurrence was found more prevalent for those male patients who were having the habit of either tobacco chewing, smoking or drinking alcohols. This work further reports in-situ LIBS study of deciduous tooth and in-vivo LIBS study of human nail.

Simulating Extraterrestrial Ices in the Laboratory

NASA Astrophysics Data System (ADS)

Berisford, D. F.; Carey, E. M.; Hand, K. P.; Choukroun, M.

2017-12-01

Several ongoing experiments at JPL attempt to simulate the ice environment for various regimes associated with icy moons. The Europa Penitent Ice Experiment (EPIX) simulates the surface environment of an icy moon, to investigate the physics of ice surface morphology growth. This experiment features half-meter-scale cryogenic ice samples, cryogenic radiative sink environment, vacuum conditions, and diurnal cycling solar simulation. The experiment also includes several smaller fixed-geometry vacuum chambers for ice simulation at Earth-like and intermediate temperature and vacuum conditions for development of surface morphology growth scaling relations. Additionally, an ice cutting facility built on a similar platform provides qualitative data on the mechanical behavior of cryogenic ice with impurities under vacuum, and allows testing of ice cutting/sampling tools relevant for landing spacecraft. A larger cutting facility is under construction at JPL, which will provide more quantitative data and allow full-scale sampling tool tests. Another facility, the JPL Ice Physics Laboratory, features icy analog simulant preparation abilities that range icy solar system objects such as Mars, Ceres and the icy satellites of Saturn and Jupiter. In addition, the Ice Physics Lab has unique facilities for Icy Analog Tidal Simulation and Rheological Studies of Cryogenic Icy Slurries, as well as equipment to perform thermal and mechanical properties testing on icy analog materials and their response to sinusoidal tidal stresses.

Detection of glass particles on bone lesions using SEM-EDS.

PubMed

Montoriol, Romain; Guilbeau-Frugier, Céline; Chantalat, Elodie; Roumiguié, Mathieu; Delisle, Marie-Bernadette; Payré, Bruno; Telmon, Norbert; Savall, Frédéric

2017-09-01

The problem of identifying the wounding agent in forensic cases is recurrent. Moreover, when several tools are involved, distinguishing the origin of lesions can be difficult. Scanning electron microscopy (SEM)/energy dispersive X-ray analysis (EDS) equipment is increasingly available to the scientific and medical community, and some studies have reported its use in forensic anthropology. However, at our knowledge, no study has reported the use of SEM-EDS in forensic cases involving glass tools, whether in case reports or experiments. We performed an experimental study on human rib fragments, on which we manually created wounds using fragments of window and mirror glass. SEM-EDS was executed on samples without any further preparation on low vacuum mode, then on the same samples after defleshing them completely by boiling them. Window and mirror glass particles were detected on experimental wounds. Both had silica in their spectra, and the opaque side of the mirror contained titanium, allowing for their identification. Boiling and defleshing the bone samples involved a loss of information in terms of the number of wounds detected as positive for glass particles and in the number of glass particles detected, for both window and mirror glass. We suggest the analysis of wounds with suspected glass particles using low vacuum mode and with no defleshment by boiling.

Effect of Low Nickel Dopant on Torque Transducer Response Function in High-Chromium Content ESR Stainless Tool Steels

NASA Astrophysics Data System (ADS)

Wiewel, Joseph L.; Hecox, Bryan G.; Orris, Jason T.; Boley, Mark S.

2007-03-01

The change in magnetoelastic torque transducer response was investigated as a low nickel content (up to 0.2%) is alloyed into an ESR (Electro-Slag-Refining) stainless tool steel with a chromium content of around 13%, which our previous studies have proven to be the ideal level of chromium content for optimal transducer performance. Two separate hollow steel 3/4-inch diameter shafts were prepared from ESR 416 and ESR 420 steel, respectively, the first having no nickel content and the second having 0.2% nickel content. The heat treatment of these steels consisted of a hardening process conducted in a helium atmosphere at 1038^oC, followed by an annealing at 871^oC for 5h and a 15^oC cool down rate. Prior and subsequent to the heat treatment processes, the circumferential and axial magnetic hysteresis properties of the samples were measured and their external field signals were mapped over the magnetically polarized regions both with and without applied shear stress up to 2500 psi on the samples. It was found that the effect of the low nickel dopant was to improve torque transducer sensitivity and linearity, but heat treatment worsened the performance of both samples.

Improving tritium exposure reconstructions using accelerator mass spectrometry

PubMed Central

Hunt, J. R.; Vogel, J. S.; Knezovich, J. P.

2010-01-01

Direct measurement of tritium atoms by accelerator mass spectrometry (AMS) enables rapid low-activity tritium measurements from milligram-sized samples and permits greater ease of sample collection, faster throughput, and increased spatial and/or temporal resolution. Because existing methodologies for quantifying tritium have some significant limitations, the development of tritium AMS has allowed improvements in reconstructing tritium exposure concentrations from environmental measurements and provides an important additional tool in assessing the temporal and spatial distribution of chronic exposure. Tritium exposure reconstructions using AMS were previously demonstrated for a tree growing on known levels of tritiated water and for trees exposed to atmospheric releases of tritiated water vapor. In these analyses, tritium levels were measured from milligram-sized samples with sample preparation times of a few days. Hundreds of samples were analyzed within a few months of sample collection and resulted in the reconstruction of spatial and temporal exposure from tritium releases. Although the current quantification limit of tritium AMS is not adequate to determine natural environmental variations in tritium concentrations, it is expected to be sufficient for studies assessing possible health effects from chronic environmental tritium exposure. PMID:14735274

The ExoMars Sample Preparation and Distribution System

NASA Astrophysics Data System (ADS)

Schulte, Wolfgang; Hofmann, Peter; Baglioni, Pietro; Richter, Lutz; Redlich, . Daniel; Notarnicola, Marco; Durrant, Stephen

2012-07-01

The Sample Preparation and Distribution System (SPDS) is a key element of the ESA ExoMars Rover. It is a set of complex mechanisms designed to receive Mars soil samples acquired from the subsurface with a drill, to crush them and to distribute the obtained soil powder to the scientific instruments of the `Pasteur Payload', in the Rover Analytical Laboratory (ALD). In particular, the SPDS consists of: (1) a Core Sample Handling System (CSHS), including a Core Sample Transportation Mechanism (CSTM) and a Blank Sample Dispenser; (2) a Crushing Station (CS); (3) a Powder Sample Dosing and Distribution System (PSDDS); and (4) a Powder Sample Handling System (PSHS) which is a carousel carrying pyrolysis ovens, a re-fillable sample container and a tool to flatten the powder sample surface. Kayser-Threde has developed, undercontract with the ExoMars prime contractor Thales Alenia Space Italy, breadboards and an engineering model of the SPDS mechanisms. Tests of individual mechanisms, namely the CSTM, CS and PSDDS were conducted both in laboratory ambient conditions and in a simulated Mars environment, using dedicated facilities. The SPDS functionalities and performances were measured and evaluated. In the course of 2011 the SPDS Dosing Station (part of the PSDDS) was also tested in simulated Mars gravity conditions during a parabolic flight campaign. By the time of the conference, an elegant breadboard of the Powder Sample Handling System will have been built and tested. The next step, planned by mid of 2012, will be a complete end-to-end test of the sample handling and processing chain, combining all four SPDS mechanisms. The possibility to verify interface and operational aspects between the SPDS and the ALD scientific instruments using the available instruments breadboards with the end-to-end set-up is currently being evaluated. This paper illustrates the most recent design status of the SPDS mechanisms, summarizes the test results and highlights future development activities, including potential involvement of the ExoMars science experiments.

Metadynamics for training neural network model chemistries: A competitive assessment

NASA Astrophysics Data System (ADS)

Herr, John E.; Yao, Kun; McIntyre, Ryker; Toth, David W.; Parkhill, John

2018-06-01

Neural network model chemistries (NNMCs) promise to facilitate the accurate exploration of chemical space and simulation of large reactive systems. One important path to improving these models is to add layers of physical detail, especially long-range forces. At short range, however, these models are data driven and data limited. Little is systematically known about how data should be sampled, and "test data" chosen randomly from some sampling techniques can provide poor information about generality. If the sampling method is narrow, "test error" can appear encouragingly tiny while the model fails catastrophically elsewhere. In this manuscript, we competitively evaluate two common sampling methods: molecular dynamics (MD), normal-mode sampling, and one uncommon alternative, Metadynamics (MetaMD), for preparing training geometries. We show that MD is an inefficient sampling method in the sense that additional samples do not improve generality. We also show that MetaMD is easily implemented in any NNMC software package with cost that scales linearly with the number of atoms in a sample molecule. MetaMD is a black-box way to ensure samples always reach out to new regions of chemical space, while remaining relevant to chemistry near kbT. It is a cheap tool to address the issue of generalization.

Preparing Effective Special Education Teachers. What Works for Special-Needs Learners Series

ERIC Educational Resources Information Center

Mamlin, Nancy

2012-01-01

What tools are in the toolkit of an excellent special educator, and how can teacher preparation programs provide these tools in the most efficient, effective way possible? This practical, clearly written book is grounded in current research and policy as well as the author's extensive experience as a teacher educator. It identifies what special…

Thermal effects of the Er:YAG laser on a simulated dental pulp: a quantitative evaluation of the effects of a water spray.

PubMed

Attrill, D C; Davies, R M; King, T A; Dickinson, M R; Blinkhorn, A S

2004-01-01

To quantify the temperature increments in a simulated dental pulp following irradiation with an Er:YAG laser, and to compare those increments when the laser is applied with and without water spray. Two cavities were prepared on either the buccal or lingual aspect of sound extracted teeth using the laser. One cavity was prepared with water spray, the other without and the order of preparation randomised. Identical preparation parameters were used for both cavities. Temperature increments were measured in the pulp chamber using a calibrated thermocouple and a novel pulp simulant. Maximum increments were 4.0 degrees C (water) and 24.7 degrees C (no water). Water was shown to be highly significant in reducing the overall temperature increments in all cases (p<0.001; paired t-test). None of the samples prepared up to a maximum of 135 J cumulative energy prepared with water spray exceeded that threshold at which pulpal damage can be considered to occur. Only 25% of those prepared without water spray remained below this threshold. Extrapolation of the figures suggests probably tolerable limits of continuous laser irradiation with water in excess to 160 J. With the incorporation of small breaks in the continuity of laser irradiation that occur in the in vivo situation, the cumulative energy dose tolerated by the pulp should far exceed these figures. The Er:YAG laser must be used in conjunction with water during cavity preparation. As such it should be considered as an effective tool for clinical use based on predicted pulpal responses to thermal stimuli.

An optimised protocol for molecular identification of Eimeria from chickens☆

PubMed Central

Kumar, Saroj; Garg, Rajat; Moftah, Abdalgader; Clark, Emily L.; Macdonald, Sarah E.; Chaudhry, Abdul S.; Sparagano, Olivier; Banerjee, Partha S.; Kundu, Krishnendu; Tomley, Fiona M.; Blake, Damer P.

2014-01-01

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples. PMID:24138724

Concentration of polycyclic aromatic hydrocarbons in water samples from different stages of treatment

NASA Astrophysics Data System (ADS)

Pogorzelec, Marta; Piekarska, Katarzyna

2017-11-01

The aim of this study was to analyze the presence and concentration of selected polycyclic aromatic hydrocarbons in water samples from different stages of treatment and to verify the usefulness of semipermeable membrane devices for analysis of drinking water. For this purpose, study was conducted for a period of 5 months. Semipermeable membrane devices were deployed in a surface water treatment plant located in Lower Silesia (Poland). To determine the effect of water treatment on concentration of PAHs, three sampling places were chosen: raw water input, stream of water just before disinfection and treated water output. After each month of sampling SPMDs were changed for fresh ones and prepared for further analysis. Concentrations of fifteen polycyclic aromatic hydrocarbons were determined by high performance liquid chromatography (HPLC). Presented study indicates that the use of semipermeable membrane devices can be an effective tool for the analysis of aquatic environment, including monitoring of drinking water, where organic micropollutants are present at very low concentrations.

Fast quantification of bovine milk proteins employing external cavity-quantum cascade laser spectroscopy.

PubMed

Schwaighofer, Andreas; Kuligowski, Julia; Quintás, Guillermo; Mayer, Helmut K; Lendl, Bernhard

2018-06-30

Analysis of proteins in bovine milk is usually tackled by time-consuming analytical approaches involving wet-chemical, multi-step sample clean-up procedures. The use of external cavity-quantum cascade laser (EC-QCL) based IR spectroscopy was evaluated as an alternative screening tool for direct and simultaneous quantification of individual proteins (i.e. casein and β-lactoglobulin) and total protein content in commercial bovine milk samples. Mid-IR spectra of protein standard mixtures were used for building partial least squares (PLS) regression models. A sample set comprising different milk types (pasteurized; differently processed extended shelf life, ESL; ultra-high temperature, UHT) was analysed and results were compared to reference methods. Concentration values of the QCL-IR spectroscopy approach obtained within several minutes are in good agreement with reference methods involving multiple sample preparation steps. The potential application as a fast screening method for estimating the heat load applied to liquid milk is demonstrated. Copyright © 2018 Elsevier Ltd. All rights reserved.

Fluorescence suppression using wavelength modulated Raman spectroscopy in fiber-probe-based tissue analysis.

PubMed

Praveen, Bavishna B; Ashok, Praveen C; Mazilu, Michael; Riches, Andrew; Herrington, Simon; Dholakia, Kishan

2012-07-01

In the field of biomedical optics, Raman spectroscopy is a powerful tool for probing the chemical composition of biological samples. In particular, fiber Raman probes play a crucial role for in vivo and ex vivo tissue analysis. However, the high-fluorescence background typically contributed by the auto fluorescence from both a tissue sample and the fiber-probe interferes strongly with the relatively weak Raman signal. Here we demonstrate the implementation of wavelength-modulated Raman spectroscopy (WMRS) to suppress the fluorescence background while analyzing tissues using fiber Raman probes. We have observed a significant signal-to-noise ratio enhancement in the Raman bands of bone tissue, which have a relatively high fluorescence background. Implementation of WMRS in fiber-probe-based bone tissue study yielded usable Raman spectra in a relatively short acquisition time (∼30  s), notably without any special sample preparation stage. Finally, we have validated its capability to suppress fluorescence on other tissue samples such as adipose tissue derived from four different species.

Developing an educational video on lung lobectomy for the general surgery resident.

PubMed

Hayden, Emily L; Seagull, F Jacob; Reddy, Rishindra M

2015-06-15

The educational resources available to general surgery residents preparing for complex thoracic surgeries vary greatly in content and target audience. We hypothesized that the preparatory resources could be improved in both efficiency of use and targeting. A formal needs analysis was performed to determine residents' knowledge gaps and desired format and/or content of an educational tool while preparing for their first lung resections. The results of the needs assessment then guided the creation of a 20-min video tool. The video was evaluated by a focus group of experts for appropriateness to the target audience, ease of use, and relevance. The needs assessment illustrated that residents feel there is a paucity of appropriate resources available to them while preparing for the lung resection procedure; 82% of respondents felt that easy-to-use and concise resources on the lobectomy procedure were either "not at all" or "somewhat" accessible. Residents reported that video was their preferred format for a learning tool overall and identified a broad spectrum of most challenging procedural aspects. These results were used to guide the creation of a 20-min video tool. A focus group validated the efficacy and appropriateness of the video. Targeted and efficient tools for residents preparing for complex subspecialty procedures are needed and valued. These results clearly encourage further work in the creation of focused educational tools for surgical residents, especially in the format of short video overviews. Copyright © 2015 Elsevier Inc. All rights reserved.

Characterizing millisecond intermediates in hemoproteins using rapid-freeze-quench resonance Raman spectroscopy.

PubMed

Matsumura, Hirotoshi; Moënne-Loccoz, Pierre

2014-01-01

The combination of rapid freeze quenching (RFQ) with resonance Raman (RR) spectroscopy represents a unique tool with which to investigate the nature of short-lived intermediates formed during the enzymatic reactions of metalloproteins. Commercially available equipment allows trapping of intermediates within a millisecond to second time scale for low-temperature RR analysis resulting in the direct detection of metal-ligand vibrations and porphyrin skeletal vibrations in hemoproteins. This chapter briefly discusses RFQ-RR studies carried out previously in our laboratory and presents, as a practical example, protocols for the preparation of RFQ samples of the reaction of metmyoglobin with nitric oxide (NO) under anaerobic conditions. Also described are important controls and practical procedures for the analysis of these samples by low-temperature RR spectroscopy.

The possibility of multi-layer nanofabrication via atomic force microscope-based pulse electrochemical nanopatterning

NASA Astrophysics Data System (ADS)

Kim, Uk Su; Morita, Noboru; Lee, Deug Woo; Jun, Martin; Park, Jeong Woo

2017-05-01

Pulse electrochemical nanopatterning, a non-contact scanning probe lithography process using ultrashort voltage pulses, is based primarily on an electrochemical machining process using localized electrochemical oxidation between a sharp tool tip and the sample surface. In this study, nanoscale oxide patterns were formed on silicon Si (100) wafer surfaces via electrochemical surface nanopatterning, by supplying external pulsed currents through non-contact atomic force microscopy. Nanoscale oxide width and height were controlled by modulating the applied pulse duration. Additionally, protruding nanoscale oxides were removed completely by simple chemical etching, showing a depressed pattern on the sample substrate surface. Nanoscale two-dimensional oxides, prepared by a localized electrochemical reaction, can be defined easily by controlling physical and electrical variables, before proceeding further to a layer-by-layer nanofabrication process.

High-Throughput Industrial Coatings Research at The Dow Chemical Company.

PubMed

Kuo, Tzu-Chi; Malvadkar, Niranjan A; Drumright, Ray; Cesaretti, Richard; Bishop, Matthew T

2016-09-12

At The Dow Chemical Company, high-throughput research is an active area for developing new industrial coatings products. Using the principles of automation (i.e., using robotic instruments), parallel processing (i.e., prepare, process, and evaluate samples in parallel), and miniaturization (i.e., reduce sample size), high-throughput tools for synthesizing, formulating, and applying coating compositions have been developed at Dow. In addition, high-throughput workflows for measuring various coating properties, such as cure speed, hardness development, scratch resistance, impact toughness, resin compatibility, pot-life, surface defects, among others have also been developed in-house. These workflows correlate well with the traditional coatings tests, but they do not necessarily mimic those tests. The use of such high-throughput workflows in combination with smart experimental designs allows accelerated discovery and commercialization.

GIS Tools to Estimate Average Annual Daily Traffic

DOT National Transportation Integrated Search

2012-06-01

This project presents five tools that were created for a geographical information system to estimate Annual Average Daily : Traffic using linear regression. Three of the tools can be used to prepare spatial data for linear regression. One tool can be...

Tracing origins of complex pharmaceutical preparations using surface desorption atmospheric pressure chemical ionization mass spectrometry.

PubMed

Zhang, Xinglei; Jia, Bin; Huang, Keke; Hu, Bin; Chen, Rong; Chen, Huanwen

2010-10-01

A novel strategy to trace the origins of commercial pharmaceutical products has been developed based on the direct chemical profiling of the pharmaceutical products by surface desorption atmospheric pressure chemical ionization mass spectrometry (DAPCI-MS). Besides the unambiguous identification of active drug components, various compounds present in the matrixes are simultaneously detected without sample pretreatment, providing valuable information for drug quality control and origin differentiation. Four sources of commercial amoxicillin products made by different manufacturers have been successfully differentiated. This strategy has been extended to secerning six sources of Liuwei Dihuang Teapills, which are herbal medicine preparations with extremely complex matrixes. The photolysis status of chemical drug products and the inferior natural herd medicine products prepared with different processes (e.g., extra heating) were also screened using the method reported here. The limit of detection achieved in the MS/MS experiments was estimated to be 1 ng/g for amoxicillin inside the capsule product. Our experimental data demonstrate that DAPCI-MS is a useful tool for rapid pharmaceutical analysis, showing promising perspectives for tracking the entire pharmaceutical supply chain to prevent counterfeit intrusions.

Imaging plasmodesmata with high-resolution scanning electron microscopy.

PubMed

Barton, Deborah A; Overall, Robyn L

2015-01-01

High-resolution scanning electron microscopy (HRSEM) is an effective tool to investigate the distribution of plasmodesmata within plant cell walls as well as to probe their complex, three-dimensional architecture. It is a useful alternative to traditional transmission electron microscopy (TEM) in which plasmodesmata are sectioned to reveal their internal substructures. Benefits of adopting an HRSEM approach to studies of plasmodesmata are that the specimen preparation methods are less complex and time consuming than for TEM, many plasmodesmata within a large region of tissue can be imaged in a single session, and three-dimensional information is readily available without the need for reconstructing TEM serial sections or employing transmission electron tomography, both of which are lengthy processes. Here we describe methods to prepare plant samples for HRSEM using pre- or postfixation extraction of cellular material in order to visualize plasmodesmata embedded within plant cell walls.

Elemental Analysis in Biological Matrices Using ICP-MS.

PubMed

Hansen, Matthew N; Clogston, Jeffrey D

2018-01-01

The increasing exploration of metallic nanoparticles for use as cancer therapeutic agents necessitates a sensitive technique to track the clearance and distribution of the material once introduced into a living system. Inductively coupled plasma mass spectrometry (ICP-MS) provides a sensitive and selective tool for tracking the distribution of metal components from these nanotherapeutics. This chapter presents a standardized method for processing biological matrices, ensuring complete homogenization of tissues, and outlines the preparation of appropriate standards and controls. The method described herein utilized gold nanoparticle-treated samples; however, the method can easily be applied to the analysis of other metals.

A novel strategy using MASCOT Distiller for analysis of cleavable isotope-coded affinity tag data to quantify protein changes in plasma.

PubMed

Leung, Kit-Yi; Lescuyer, Pierre; Campbell, James; Byers, Helen L; Allard, Laure; Sanchez, Jean-Charles; Ward, Malcolm A

2005-08-01

A novel strategy consisting of cleavable Isotope-Coded Affinity Tag (cICAT) combined with MASCOT Distiller was evaluated as a tool for the quantification of proteins in "abnormal" patient plasma, prepared by pooling samples from patients with acute stroke. Quantification of all light and heavy cICAT-labelled peptide ion pairs was obtained using MASCOT Distiller combined with a proprietary software. Peptides displaying differences were selected for identification by MS. These preliminary results show the promise of our approach to identify potential biomarkers.

Proteomics of filamentous fungi.

PubMed

Kim, Yonghyun; Nandakumar, M P; Marten, Mark R

2007-09-01

Proteomic analysis, defined here as the global assessment of cellular proteins expressed in a particular biological state, is a powerful tool that can provide a systematic understanding of events at the molecular level. Proteomic studies of filamentous fungi have only recently begun to appear in the literature, despite the prevalence of these organisms in the biotechnology industry, and their importance as both human and plant pathogens. Here, we review recent publications that have used a proteomic approach to develop a better understanding of filamentous fungi, highlighting sample preparation methods and whole-cell cytoplasmic proteomics, as well as subproteomics of cell envelope, mitochondrial and secreted proteins.

Conversion of NIMROD simulation results for graphical analysis using VisIt

DOE Office of Scientific and Technical Information (OSTI.GOV)

Romero-Talamas, C A

Software routines developed to prepare NIMROD [C. R. Sovinec et al., J. Comp. Phys. 195, 355 (2004)] results for three-dimensional visualization from simulations of the Sustained Spheromak Physics Experiment (SSPX ) [E. B. Hooper et al., Nucl. Fusion 39, 863 (1999)] are presented here. The visualization is done by first converting the NIMROD output to a format known as legacy VTK and then loading it to VisIt, a graphical analysis tool that includes three-dimensional rendering and various mathematical operations for large data sets. Sample images obtained from the processing of NIMROD data with VisIt are included.

A "three-in-one" sample preparation method for simultaneous determination of B-group water-soluble vitamins in infant formula using VitaFast(®) kits.

PubMed

Zhang, Heng; Lan, Fang; Shi, Yupeng; Wan, Zhi-Gang; Yue, Zhen-Feng; Fan, Fang; Lin, Yan-Kui; Tang, Mu-Jin; Lv, Jing-Zhang; Xiao, Tan; Yi, Changqing

2014-06-15

VitaFast(®) test kits designed for the microbiological assay in microtiter plate format can be applied to quantitative determination of B-group water-soluble vitamins such as vitamin B12, folic acid and biotin, et al. Compared to traditional microbiological methods, VitaFast(®) kits significantly reduce sample processing time and provide greater reliability, higher productivity and better accuracy. Recently, simultaneous determination of vitamin B12, folic acid and biotin in one sample is urgently required when evaluating the quality of infant formulae in our practical work. However, the present sample preparation protocols which are developed for individual test systems, are incompatible with simultaneous determination of several analytes. To solve this problem, a novel "three-in-one" sample preparation method is herein developed for simultaneous determination of B-group water-soluble vitamins using VitaFast(®) kits. The performance of this novel "three-in-one" sample preparation method was systematically evaluated through comparing with individual sample preparation protocols. The experimental results of the assays which employed "three-in-one" sample preparation method were in good agreement with those obtained from conventional VitaFast(®) extraction methods, indicating that the proposed "three-in-one" sample preparation method is applicable to the present three VitaFast(®) vitamin test systems, thus offering a promising alternative for the three independent sample preparation methods. The proposed new sample preparation method will significantly improve the efficiency of infant formulae inspection. Copyright © 2013 Elsevier Ltd. All rights reserved.

The development of plant food processing in the Levant: insights from use-wear analysis of Early Epipalaeolithic ground stone tools

PubMed Central

Dubreuil, Laure; Nadel, Dani

2015-01-01

In recent years, the study of percussive, pounding and grinding tools has provided new insights into human evolution, more particularly regarding the development of technology enabling the processing and exploitation of plant resources. Some of these studies focus on early evidence for flour production, an activity frequently perceived as an important step in the evolution of plant exploitation. The present paper investigates plant food preparation in mobile hunter-gatherer societies from the Southern Levant. The analysis consists of a use-wear study of 18 tools recovered from Ohalo II, a 23 000-year-old site in Israel showing an exceptional level of preservation. Our sample includes a slab previously interpreted as a lower implement used for producing flour, based on the presence of cereal starch residues. The use-wear data we have obtained provide crucial information about the function of this and other percussive tools at Ohalo II, as well as on investment in tool manufacture, discard strategies and evidence for plant processing in the Late Pleistocene. The use-wear analysis indicates that the production of flour was a sporadic activity at Ohalo II, predating by thousands of years the onset of routine processing of plant foods. PMID:26483535

The development of plant food processing in the Levant: insights from use-wear analysis of Early Epipalaeolithic ground stone tools.

PubMed

Dubreuil, Laure; Nadel, Dani

2015-11-19

In recent years, the study of percussive, pounding and grinding tools has provided new insights into human evolution, more particularly regarding the development of technology enabling the processing and exploitation of plant resources. Some of these studies focus on early evidence for flour production, an activity frequently perceived as an important step in the evolution of plant exploitation. The present paper investigates plant food preparation in mobile hunter-gatherer societies from the Southern Levant. The analysis consists of a use-wear study of 18 tools recovered from Ohalo II, a 23 000-year-old site in Israel showing an exceptional level of preservation. Our sample includes a slab previously interpreted as a lower implement used for producing flour, based on the presence of cereal starch residues. The use-wear data we have obtained provide crucial information about the function of this and other percussive tools at Ohalo II, as well as on investment in tool manufacture, discard strategies and evidence for plant processing in the Late Pleistocene. The use-wear analysis indicates that the production of flour was a sporadic activity at Ohalo II, predating by thousands of years the onset of routine processing of plant foods. © 2015 The Author(s).

Standardized Sample Preparation Using a Drop-on-Demand Printing Platform

DTIC Science & Technology

2013-05-07

successful and robust methodology for energetic sample preparation. Keywords: drop-on-demand; inkjet printing; sample preparation OPEN ACCESS...on a similar length scale. Recently, drop-on-demand inkjet printing technology has emerged as an effective approach to produce test materials to...which most of the material is concentrated along the edges, samples prepared using drop-on-demand inkjet technology demonstrate excellent uniform

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haudebourg, Raphael; Fichet, Pascal; Goutelard, Florence

The detection (location and quantification) of nuclear facilities to be dismantled possible contamination with low-range particles emitters ({sup 3}H, other low-energy β emitters, a emitters) remains a tedious and expensive task. Indeed, usual remote counters show a too low sensitivity to these non-penetrating radiations, while conventional wipe tests are irrelevant for fixed radioactivity evaluation. The only method to accurately measure activity levels consists in sampling and running advanced laboratory analyses (spectroscopy, liquid scintillation counting, pyrolysis...). Such measurements generally induce sample preparation, waste production (destructive analyses, solvents), nuclear material transportation, long durations, and significant labor mobilization. Therefore, the search for themore » limitation of their number and cost easily conflicts with the necessity to perform a dense screening for sampling (to maximize the representativeness of the samples), in installations of thousands of square meters (floors, wells, ceilings), plus furniture, pipes, and other wastes. To overcome this contradiction, Digital Autoradiography (D. A.) was re-routed from bio molecular research to radiological mapping of nuclear installations under dismantling and to waste and sample analysis. After in-situ exposure to the possibly-contaminated areas to investigate, commercial reusable radiosensitive phosphor screens (of a few 100 cm{sup 2}) were scanned in the proper laboratory device and sharp quantitative images of the radioactivity could be obtained. The implementation of geostatistical tools in the data processing software enabled the exhaustive characterization of concrete floors at a rate of 2 weeks / 100 m{sup 2}, at lowest costs. Various samples such as drilled cores, or tank and wood pieces, were also successfully evaluated with this method, for decisive results. Thanks to the accurate location of potential contamination spots, this approach ensures relevant and representative sampling for further laboratory analyses and should be inserted in the range of common tools used in dismantling. (authors)« less

Can cloud point-based enrichment, preservation, and detection methods help to bridge gaps in aquatic nanometrology?

PubMed

Duester, Lars; Fabricius, Anne-Lena; Jakobtorweihen, Sven; Philippe, Allan; Weigl, Florian; Wimmer, Andreas; Schuster, Michael; Nazar, Muhammad Faizan

2016-11-01

Coacervate-based techniques are intensively used in environmental analytical chemistry to enrich and extract different kinds of analytes. Most methods focus on the total content or the speciation of inorganic and organic substances. Size fractionation is less commonly addressed. Within coacervate-based techniques, cloud point extraction (CPE) is characterized by a phase separation of non-ionic surfactants dispersed in an aqueous solution when the respective cloud point temperature is exceeded. In this context, the feature article raises the following question: May CPE in future studies serve as a key tool (i) to enrich and extract nanoparticles (NPs) from complex environmental matrices prior to analyses and (ii) to preserve the colloidal status of unstable environmental samples? With respect to engineered NPs, a significant gap between environmental concentrations and size- and element-specific analytical capabilities is still visible. CPE may support efforts to overcome this "concentration gap" via the analyte enrichment. In addition, most environmental colloidal systems are known to be unstable, dynamic, and sensitive to changes of the environmental conditions during sampling and sample preparation. This delivers a so far unsolved "sample preparation dilemma" in the analytical process. The authors are of the opinion that CPE-based methods have the potential to preserve the colloidal status of these instable samples. Focusing on NPs, this feature article aims to support the discussion on the creation of a convention called the "CPE extractable fraction" by connecting current knowledge on CPE mechanisms and on available applications, via the uncertainties visible and modeling approaches available, with potential future benefits from CPE protocols.

40 CFR 51.5 - What tools are available to help prepare and report emissions data?

Code of Federal Regulations, 2010 CFR

2010-07-01

... prepare and report emissions data? 51.5 Section 51.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS REQUIREMENTS FOR PREPARATION, ADOPTION, AND SUBMITTAL OF IMPLEMENTATION... are available to help prepare and report emissions data? (a) We urge your state to use estimation...

40 CFR 51.5 - What tools are available to help prepare and report emissions data?

Code of Federal Regulations, 2011 CFR

2011-07-01

... prepare and report emissions data? 51.5 Section 51.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS REQUIREMENTS FOR PREPARATION, ADOPTION, AND SUBMITTAL OF IMPLEMENTATION... are available to help prepare and report emissions data? (a) We urge your state to use estimation...

Advanced Pressure Coring System for Deep Earth Sampling (APRECOS)

NASA Astrophysics Data System (ADS)

Anders, E.; Rothfuss, M.; Müller, W. H.

2009-04-01

Nowadays the recovery of cores from boreholes is a standard operation. However, during that process the mechanical, physical, and chemical properties as well as living conditions for microorganisms are significantly altered. In-situ sampling is one approach to overcome the severe scientific limitations of conventional, depressurized core investigations by recovering, processing, and conducting experiments in the laboratory, while maintaining unchanged environmental parameters. The most successful equipment today is the suite of tools developed within the EU funded projects HYACE (Hydrate Autoclave Coring Equipment) and HYACINTH (Deployment of HYACE tools In New Tests on Hydrates) between 1997 and 2005. Within several DFG (German Research Foundation) projects the Technical University Berlin currently works on concepts to increase the present working pressure of 250 bar as well as to reduce logistical and financial expenses by merging redundant and analogous procedures and scaling down the considerable size of key components. It is also proposed to extend the range of applications for the wireline rotary pressure corer and the sub-sampling and transfer system to all types of soil conditions (soft to highly-consolidated). New modifications enable the tools to be used in other pressure related fields of research, such as unconventional gas exploration (coal-bed methane, tight gas, gas hydrate), CO2 sequestration, and microbiology of the deep biosphere. Expedient enhancement of an overall solution for pressure core retrieval, process and investigation will open the way for a complete on-site, all-purpose, in-situ equipment. The advanced assembly would allow for executing the whole operation sequences of coring, non-destructive measurement, sub-sampling and transfer into storage, measurement and transportation chambers, all in sterile, anaerobic conditions, and without depressurisation in quick succession. Extensive post-cruise handling and interim storage would be dispensable. The complete core processing and preparation of in-situ sample sections for worldwide shipping could be conducted within hours after retrieval.

Full genome virus detection in fecal samples using sensitive nucleic acid preparation, deep sequencing, and a novel iterative sequence classification algorithm.

PubMed

Cotten, Matthew; Oude Munnink, Bas; Canuti, Marta; Deijs, Martin; Watson, Simon J; Kellam, Paul; van der Hoek, Lia

2014-01-01

We have developed a full genome virus detection process that combines sensitive nucleic acid preparation optimised for virus identification in fecal material with Illumina MiSeq sequencing and a novel post-sequencing virus identification algorithm. Enriched viral nucleic acid was converted to double-stranded DNA and subjected to Illumina MiSeq sequencing. The resulting short reads were processed with a novel iterative Python algorithm SLIM for the identification of sequences with homology to known viruses. De novo assembly was then used to generate full viral genomes. The sensitivity of this process was demonstrated with a set of fecal samples from HIV-1 infected patients. A quantitative assessment of the mammalian, plant, and bacterial virus content of this compartment was generated and the deep sequencing data were sufficient to assembly 12 complete viral genomes from 6 virus families. The method detected high levels of enteropathic viruses that are normally controlled in healthy adults, but may be involved in the pathogenesis of HIV-1 infection and will provide a powerful tool for virus detection and for analyzing changes in the fecal virome associated with HIV-1 progression and pathogenesis.

Full Genome Virus Detection in Fecal Samples Using Sensitive Nucleic Acid Preparation, Deep Sequencing, and a Novel Iterative Sequence Classification Algorithm

PubMed Central

Cotten, Matthew; Oude Munnink, Bas; Canuti, Marta; Deijs, Martin; Watson, Simon J.; Kellam, Paul; van der Hoek, Lia

2014-01-01

We have developed a full genome virus detection process that combines sensitive nucleic acid preparation optimised for virus identification in fecal material with Illumina MiSeq sequencing and a novel post-sequencing virus identification algorithm. Enriched viral nucleic acid was converted to double-stranded DNA and subjected to Illumina MiSeq sequencing. The resulting short reads were processed with a novel iterative Python algorithm SLIM for the identification of sequences with homology to known viruses. De novo assembly was then used to generate full viral genomes. The sensitivity of this process was demonstrated with a set of fecal samples from HIV-1 infected patients. A quantitative assessment of the mammalian, plant, and bacterial virus content of this compartment was generated and the deep sequencing data were sufficient to assembly 12 complete viral genomes from 6 virus families. The method detected high levels of enteropathic viruses that are normally controlled in healthy adults, but may be involved in the pathogenesis of HIV-1 infection and will provide a powerful tool for virus detection and for analyzing changes in the fecal virome associated with HIV-1 progression and pathogenesis. PMID:24695106

Incorporating current research into formal higher education settings using Astrobites

NASA Astrophysics Data System (ADS)

Sanders, Nathan E.; Kohler, Susanna; Faesi, Chris; Villar, Ashley; Zevin, Michael

2017-10-01

A primary goal of many undergraduate- and graduate-level courses in the physical sciences is to prepare students to engage in scientific research or to prepare students for careers that leverage skillsets similar to those used by research scientists. Even for students who may not intend to pursue a career with these characteristics, exposure to the context of applications in modern research can be a valuable tool for teaching and learning. However, a persistent barrier to student participation in research is familiarity with the technical language, format, and context that academic researchers use to communicate research methods and findings with each other: the literature of the field. Astrobites, an online web resource authored by graduate students, has published brief and accessible summaries of more than 1300 articles from the astrophysical literature since its founding in 2010. This article presents three methods for introducing students at all levels within the formal higher education setting to approaches and results from modern research. For each method, we provide a sample lesson plan that integrates content and principles from Astrobites, including step-by-step instructions for instructors, suggestions for adapting the lesson to different class levels across the undergraduate and graduate spectrum, sample student handouts, and a grading rubric.

Sample preparation for SFM imaging of DNA, proteins, and DNA-protein complexes.

PubMed

Ristic, Dejan; Sanchez, Humberto; Wyman, Claire

2011-01-01

Direct imaging is invaluable for understanding the mechanism of complex genome transactions where proteins work together to organize, transcribe, replicate, and repair DNA. Scanning (or atomic) force microscopy is an ideal tool for this, providing 3D information on molecular structure at nanometer resolution from defined components. This is a convenient and practical addition to in vitro studies as readily obtainable amounts of purified proteins and DNA are required. The images reveal structural details on the size and location of DNA-bound proteins as well as protein-induced arrangement of the DNA, which are directly correlated in the same complexes. In addition, even from static images, the different forms observed and their relative distributions can be used to deduce the variety and stability of different complexes that are necessarily involved in dynamic processes. Recently available instruments that combine fluorescence with topographic imaging allow the identification of specific molecular components in complex assemblies, which broadens the applications and increases the information obtained from direct imaging of molecular complexes. We describe here basic methods for preparing samples of proteins, DNA, and complexes of the two for topographic imaging and quantitative analysis. We also describe special considerations for combined fluorescence and topographic imaging of molecular complexes.

Closed tubes preparation of graphite for high-precision AMS radiocarbon analysis

NASA Astrophysics Data System (ADS)

Hajdas, I.; Michczynska, D.; Bonani, G.; Maurer, M.; Wacker, L.

2009-04-01

Radiocarbon dating is an established tool applied in Geochronology. Technical developments of Accelerator Mass Spectrometry AMS, which allow measurements of samples containing less than 1 mg of carbon, opened opportunities for new applications. Moreover, high resolution records of the past changes require high-resolution chronologies i.e. sampling for 14C dating. In result, the field of applications is rapidly expanding and number of radiocarbon analysis is growing rapidly. Nowadays dedicated 14C AMS machines have great capacity for analysis but in order to keep up with the demand for analysis and provide the results as fast as possible a very efficient way of sample preparation is required. Sample preparation for 14C AMS analysis consists of two steps: separation of relevant carbon from the sample material (removing contamination) and preparation of graphite for AMS analysis. The last step usually involves reaction of CO2 with H2, in the presence of metal catalyst (Fe or Co) of specific mesh size heated to 550-625°C, as originally suggested by Vogel et al. (1984). Various graphitization systems have been built in order to fulfil the requirement of sample quality needed for high-precision radiocarbon data. In the early 90ties another method has been proposed (Vogel 1992) and applied by few laboratories mainly for environmental or biomedical samples. This method uses TiH2 as a source of H2 and can be easily and flexibly applied to produce graphite. Sample of CO2 is frozen in to the tube containing pre-conditioned Zn/TiH2 and Fe catalyst. Torch sealed tubes are then placed in the stepwise heated oven at 500/550°C and left to react for several hours. The greatest problem is the lack of control of the reaction completeness and considerable fractionation. However, recently reported results (Xu et al. 2007) suggest that high precision dating using graphite produced in closed tubes might be possible. We will present results of radiocarbon dating of the set of standards and secondary IAEA standards to demonstrate to what level this method can be used for high precision radiocarbon dating. References Vogel JS. 1992. Rapid Production of Graphite without Contamination for Biomedical Ams. Radiocarbon 34: 344-350. Vogel JS, Southon JR, Nelson DE, and Brown TA. 1984. Performance of Catalytically Condensed Carbon for Use in Accelerator Mass-Spectrometry. Nuclear Instruments & Methods in Physics Research Section B-Beam Interactions with Materials and Atoms 233: 289-293. Xu X, Trumbore SE, Zheng S, Southon JR, McDuffee KE, Luttgen M, and Liu JC. 2007. Modifying a sealed tube zinc reduction method for preparation of AMS graphite targets: Reducing background and attaining high precision. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms Accelerator Mass Spectrometry - Proceedings of the Tenth International Conference on Accelerator Mass Spectrometry 259: 320-329.

Focused Ion Beam (FIB) combined with SEM (FIB/SEM) and TEM: Advanced tools for nano-analysis in Geosciences

NASA Astrophysics Data System (ADS)

Wirth, R.; Morales, L. G.

2011-12-01

Focused ion beam (FIB) techniques have been successfully applied to the preparation of site-specific electron transparent membranes for transmission electron microscopy (TEM) investigations in Geosciences since several years. For example, systematic TEM studies of nano-inclusions in diamond foils prepared with FIB have improved our knowledge on diamond formation. However, FIB is not exclusively used for sample preparation for TEM application because it has been proved that one and the same TEM foil can also be used for Synchrotron IR, Synchrotron X-Ray fluorescence (XRF), scanning transmission X-Ray microscopy (STXM) and NanoSIMS analysis. In addition, FIB milling turned out to be very useful for sample preparation of Brillouin scattering experiments and has a strong potential for preparation of highly-polished, micrometer-scale samples. However, a real break through in FIB application was achieved combining a Ga-ion source of the FIB with an electron source of a scanning electron microscope (SEM) in one single instrument. The combination of FIB/SEM renders access to the third dimension of the sample possible. A cavity normal to the sample surface is sputtered with Ga-ions and this newly created inner surface is imaged with the electron beam. Alternating slicing and viewing along these cavities allow the acquisition of a sequence of images that allows the observation in 3 dimensions. Recently, this technique has been successfully applied to image the structure of grain or phase boundaries in metamorphic rocks as well as micro- and nanoporosity in shales, but its applicability goes far beyond these few examples. Combining slicing and viewing with X-Ray and electron backscatter diffraction (EBSD) analysis can provide 3D elemental mapping and 3D crystallographic orientation mapping of crystalline materials. Combined FIB/SEM devices also facilitate the preparation of substantially thinner and cleaner TEM foils (approximately 30 nm) because electron beam imaging controls the progress of the sputtering process without sputtering the sample during imaging. Electron induce sputtering is substantially smaller than ion induced sputtering. Finally, the amorphous layers created by Ga-ion sputtering and Ga-ion implantation can be removed from the foil surfaces by subsequent argon ion bombardment under a low angle of incidence and low acceleration voltage thus permitting TEM high-resolution imaging and electron energy-loss spectroscopy (EELS). Additionally, ultra-thin foils have the advantage that they are electron transparent even at 30 keV, the common operational voltage of a SEM. Therefore the electron column of the FIB/SEM system can be used as a TEM at low voltage and images can be made either in bright-field, dark field and through a high-angle annular dark field (HAADF) detector. The HAADF detector provides information about the chemical composition of the specimen with high spatial resolution because it is Z-contrast sensitive.

Segmental hair analysis for differentiation of tilidine intake from external contamination using LC-ESI-MS/MS and MALDI-MS/MS imaging.

PubMed

Poetzsch, Michael; Baumgartner, Markus R; Steuer, Andrea E; Kraemer, Thomas

2015-02-01

Segmental hair analysis has been used for monitoring changes of consumption habit of drugs. Contamination from the environment or sweat might cause interpretative problems. For this reason, hair analysis results were compared in hair samples taken 24 h and 30 days after a single tilidine dose. The 24-h hair samples already showed high concentrations of tilidine and nortilidine. Analysis of wash water from sample preparation confirmed external contamination by sweat as reason. The 30-day hair samples were still positive for tilidine in all segments. Negative wash-water analysis proved incorporation from sweat into the hair matrix. Interpretation of a forensic case was requested where two children had been administered tilidine by their nanny and tilidine/nortilidine had been detected in all hair segments, possibly indicating multiple applications. Taking into consideration the results of the present study and of MALDI-MS imaging, a single application as cause for analytical results could no longer be excluded. Interpretation of consumption behaviour of tilidine based on segmental hair analysis has to be done with caution, even after typical wash procedures during sample preparation. External sweat contamination followed by incorporation into the hair matrix can mimic chronic intake. For assessment of external contamination, hair samples should not only be collected several weeks but also one to a few days after intake. MALDI-MS imaging of single hair can be a complementary tool for interpretation. Limitations for interpretation of segmental hair analysis shown here might also be applicable to drugs with comparable physicochemical and pharmacokinetic properties. Copyright © 2014 John Wiley & Sons, Ltd.

Determination of 129I in environmental samples by AMS and NAA using an anion exchange resin disk

NASA Astrophysics Data System (ADS)

Suzuki, Takashi; Banba, Shigeru; Kitamura, Toshikatsu; Kabuto, Shoji; Isogai, Keisuke; Amano, Hikaru

2007-06-01

We have developed a new extraction method for the measurement of 129I by accelerator mass spectrometry (AMS) utilizing an anion exchange resin disk. In comparison to traditional methods such as solvent extraction and ion exchange, this method provides for simple and quick sample handling. This extraction method was tested on soil, seaweed and milk samples, but because of disk clogging, the milk samples and some of the seaweed could not be applied successfully. Using this new extraction method to prepare samples for AMS analysis produced isotope ratios of iodine in good agreement with neutron activation analysis (NAA). The disk extraction method which take half an hour is faster than previous techniques, such as solvent extraction or ion exchange which take a few hours. The combination of the disk method and the AMS measurement is a powerful tool for the determination of 129I. Furthermore, these data will be available for the environmental monitoring before and during the operation of a new nuclear fuel reprocessing plant in Japan.

A computational method for estimating the PCR duplication rate in DNA and RNA-seq experiments.

PubMed

Bansal, Vikas

2017-03-14

PCR amplification is an important step in the preparation of DNA sequencing libraries prior to high-throughput sequencing. PCR amplification introduces redundant reads in the sequence data and estimating the PCR duplication rate is important to assess the frequency of such reads. Existing computational methods do not distinguish PCR duplicates from "natural" read duplicates that represent independent DNA fragments and therefore, over-estimate the PCR duplication rate for DNA-seq and RNA-seq experiments. In this paper, we present a computational method to estimate the average PCR duplication rate of high-throughput sequence datasets that accounts for natural read duplicates by leveraging heterozygous variants in an individual genome. Analysis of simulated data and exome sequence data from the 1000 Genomes project demonstrated that our method can accurately estimate the PCR duplication rate on paired-end as well as single-end read datasets which contain a high proportion of natural read duplicates. Further, analysis of exome datasets prepared using the Nextera library preparation method indicated that 45-50% of read duplicates correspond to natural read duplicates likely due to fragmentation bias. Finally, analysis of RNA-seq datasets from individuals in the 1000 Genomes project demonstrated that 70-95% of read duplicates observed in such datasets correspond to natural duplicates sampled from genes with high expression and identified outlier samples with a 2-fold greater PCR duplication rate than other samples. The method described here is a useful tool for estimating the PCR duplication rate of high-throughput sequence datasets and for assessing the fraction of read duplicates that correspond to natural read duplicates. An implementation of the method is available at https://github.com/vibansal/PCRduplicates .

Lack of latex allergen contamination of solutions withdrawn from vials with natural rubber stoppers.

PubMed

Thomsen, D J; Burke, T G

2000-01-01

The effect on latex allergen contamination and microbial growth of a latex-allergy precaution technique for preparing injectable products was studied. The study consisted of three parts: (1) preparation of 20 samples from vials with latex-containing stoppers in accordance with conventional guidelines, (2) preparation of 20 samples in accordance with latex-allergy precaution guidelines, and (3) preparation of 5 latex-free samples and 1 latex-contaminated sample as negative and positive controls, respectively. The conventional method involved swabbing a vial top with an alcohol prep pad, puncturing the dry natural rubber stopper with an 18-gauge needle attached to a latex-free syringe, and withdrawing the contents of the vial into the syringe. The latex-allergy precaution preparation technique was similar, except that the stopper was removed before the vial contents were withdrawn. There was essentially no difference in latex allergen concentrations between the two drug preparation methods. None of the samples prepared with the standard method supported any microbial growth. One sample prepared with the latex-allergy precaution method grew bacteria. Removal of the dry rubber stopper from vials did not yield solutions with less latex allergen than solutions prepared according to conventional guidelines.

A review on recent developments in mass spectrometry instrumentation and quantitative tools advancing bacterial proteomics.

PubMed

Van Oudenhove, Laurence; Devreese, Bart

2013-06-01

Proteomics has evolved substantially since its early days, some 20 years ago. In this mini-review, we aim to provide an overview of general methodologies and more recent developments in mass spectrometric approaches used for relative and absolute quantitation of proteins. Enhancement of sensitivity of the mass spectrometers as well as improved sample preparation and protein fractionation methods are resulting in a more comprehensive analysis of proteomes. We also document some upcoming trends for quantitative proteomics such as the use of label-free quantification methods. Hopefully, microbiologists will continue to explore proteomics as a tool in their research to understand the adaptation of microorganisms to their ever changing environment. We encourage them to incorporate some of the described new developments in mass spectrometry to facilitate their analyses and improve the general knowledge of the fascinating world of microorganisms.

Opti-Tool: EPA Region 1's Stormwater Management Optimization Tool

EPA Pesticide Factsheets

Opti-Tool assists stormwater managers & consulting engineers in preparing technically sound & cost-effective watershed SW mgmt plans to achieve needed pollutant & volume reductions more affordably from developed landscapes throughout the New England Region

The Use of Lego Technologies in Elementary Teacher Preparation

NASA Astrophysics Data System (ADS)

Hadjiachilleos, Stella; Avraamidou, Lucy; Papastavrou, Stavros

2013-10-01

The need to reform science teacher preparation programs has been pointed out in research (Bryan and Abell in J Res Sci Teach 36:121-140, 1999; Bryan and Atwater in Sci Educ 8(6):821-839, 2002; Harrington and Hathaway in J Teach Educ 46(4):275-284, 1995). Science teachers are charged with the responsibility of incorporating both cognitive and non-cognitive parameters in their everyday teaching practices. This often results in their reluctance to teach science because they often lack disciplinary and/or pedagogical expertise required to promote science learning. The purpose of this study is to propose an alternative instructional approach in which Lego vehicles were used as a tool to promote pre-service elementary teachers' development and to examine whether there are non-cognitive parameters that promote or obstruct them from using Lego Technologies as a teaching tool. The context of the study was defined by a teacher preparation program of a private university in a small Mediterranean country. A sample of 28 pre-service elementary teachers, working in five 5-6-member groups were involved in scientific inquiries, during which they had to use vehicles in order to solve scientific problems related to concepts such as gear functioning, force, and motion. The nature of their cognitive engagement in the scientific inquiry process, non-cognitive parameters contributing to their cognitive engagement, and the impact of their involvement in the process on their development were examined through qualitative analysis of pre- and post-inquiry interviews, presentations of their solutions to the scientific problems and of their personal reflective journals.

[Sample preparation and bioanalysis in mass spectrometry].

PubMed

Bourgogne, Emmanuel; Wagner, Michel

2015-01-01

The quantitative analysis of compounds of clinical interest of low molecular weight (<1000 Da) in biological fluids is currently in most cases performed by liquid chromatography-mass spectrometry (LC-MS). Analysis of these compounds in biological fluids (plasma, urine, saliva, hair...) is a difficult task requiring a sample preparation. Sample preparation is a crucial part of chemical/biological analysis and in a sense is considered the bottleneck of the whole analytical process. The main objectives of sample preparation are the removal of potential interferences, analyte preconcentration, and converting (if needed) the analyte into a more suitable form for detection or separation. Without chromatographic separation, endogenous compounds, co-eluted products may affect a quantitative method in mass spectrometry performance. This work focuses on three distinct parts. First, quantitative bioanalysis will be defined, different matrices and sample preparation techniques currently used in bioanalysis by mass spectrometry of/for small molecules of clinical interest in biological fluids. In a second step the goals of sample preparation will be described. Finally, in a third step, sample preparation strategies will be made either directly ("dilute and shoot") or after precipitation.

Ambient Mass Spectrometry in Cancer Research.

PubMed

Takats, Z; Strittmatter, N; McKenzie, J S

2017-01-01

Ambient ionization mass spectrometry was developed as a sample preparation-free alternative to traditional MS-based workflows. Desorption electrospray ionization (DESI)-MS methods were demonstrated to allow the direct analysis of a broad range of samples including unaltered biological tissue specimens. In contrast to this advantageous feature, nowadays DESI-MS is almost exclusively used for sample preparation intensive mass spectrometric imaging (MSI) in the area of cancer research. As an alternative to MALDI, DESI-MSI offers matrix deposition-free experiment with improved signal in the lower (<500m/z) range. DESI-MSI enables the spatial mapping of tumor metabolism and has been broadly demonstrated to offer an alternative to frozen section histology for intraoperative tissue identification and surgical margin assessment. Rapid evaporative ionization mass spectrometry (REIMS) was developed exclusively for the latter purpose by the direct combination of electrosurgical devices and mass spectrometry. In case of the REIMS technology, aerosol particles produced by electrosurgical dissection are subjected to MS analysis, providing spectral information on the structural lipid composition of tissues. REIMS technology was demonstrated to give real-time information on the histological nature of tissues being dissected, deeming it an ideal tool for intraoperative tissue identification including surgical margin control. More recently, the method has also been used for the rapid lipidomic phenotyping of cancer cell lines as it was demonstrated in case of the NCI-60 cell line collection. © 2017 Elsevier Inc. All rights reserved.

Focused ion beam (FIB)/scanning electron microscopy (SEM) in tissue structural research.

PubMed

Leser, Vladka; Milani, Marziale; Tatti, Francesco; Tkalec, Ziva Pipan; Strus, Jasna; Drobne, Damjana

2010-10-01

The focused ion beam (FIB) and scanning electron microscope (SEM) are commonly used in material sciences for imaging and analysis of materials. Over the last decade, the combined FIB/SEM system has proven to be also applicable in the life sciences. We have examined the potential of the focused ion beam/scanning electron microscope system for the investigation of biological tissues of the model organism Porcellio scaber (Crustacea: Isopoda). Tissue from digestive glands was prepared as for conventional SEM or as for transmission electron microscopy (TEM). The samples were transferred into FIB/SEM for FIB milling and an imaging operation. FIB-milled regions were secondary electron imaged, back-scattered electron imaged, or energy dispersive X-ray (EDX) analyzed. Our results demonstrated that FIB/SEM enables simultaneous investigation of sample gross morphology, cell surface characteristics, and subsurface structures. The same FIB-exposed regions were analyzed by EDX to provide basic compositional data. When samples were prepared as for TEM, the information obtained with FIB/SEM is comparable, though at limited magnification, to that obtained from TEM. A combination of imaging, micro-manipulation, and compositional analysis appears of particular interest in the investigation of epithelial tissues, which are subjected to various endogenous and exogenous conditions affecting their structure and function. The FIB/SEM is a promising tool for an overall examination of epithelial tissue under normal, stressed, or pathological conditions.

Guide for machine tool task force members

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sutton, G.P.

1978-09-01

The purpose of the guide is to assist members of the Machine Tool Task Force (MTTF) in doing the job, preparing technical summary papers, and helping to achieve a uniform, high-quality output from this comprehensive study effort. It supplements the MTTF Plan (UCRL-52552) which contains other important information on the method of operation of MTTF that is related to the preparation of MTTF reports.

Study of temperature rises and forces on drilling bone

NASA Astrophysics Data System (ADS)

Srikanth Venkataraman, Ananya

Many different approaches have been used to prepare, store and test bone samples in order to determine its physical properties. The need to establish a standard method of specimen preparation and storage prior to experimental testing, contributed greatly to the primary part of this study. When mechanized cutting tools such as saws and drills are used, heat is produced and this raises the temperature of both the tool and the material being cut. In orthopedic and dental practices, high-speed tools are often applied to bones and teeth, and heat from these operations may result in thermal necrosis [1]. Since this can have a negative impact on the outcome of an orthopedic procedure, temperatures must be kept below the threshold that results in bone necrosis. The initial set of experiments was performed to determine the conditions under which the mechanical properties of the bone changed so as to establish the most suitable testing conditions. The hardness variation of the bone samples, under different annealing treatment conditions was used as the indicating parameter for evaluation of the change in the mechanical properties. Establishing the most appropriate section of the metacarpal sample for testing, by studying the anisotropy of the bone was another determining parameter. The second step was to examine the effects of conventional drilling as well as modulation assisted drilling on the temperature rise generated in the bone during these machining processes. In addition to this, a set of experiments were performed to ascertain how lubrication affected the temperature rise during drilling. The dynamic portions of the torque and thrust traces as well as the specific energies were compared for the different drilling conditions. Modulation showed no significant effect on the mean torque, thrust, specific energies of cutting, or temperature rise. Lubrication (flooding and misting) in both the modulation and no modulation cases drastically reduced the temperature rise during cutting, as expected. In addition to this the characteristics of the chips produced by both the methods of drilling were compared. The modulation process produced more consistent chips at the lower speed (360 rpm) and as the speed was increased to 3000 rpm the chip formation was similar to the no modulation drilling condition at the same feed rates. A brief study on the histological changes due to drilling was also performed.

Small RNA Library Preparation Method for Next-Generation Sequencing Using Chemical Modifications to Prevent Adapter Dimer Formation.

PubMed

Shore, Sabrina; Henderson, Jordana M; Lebedev, Alexandre; Salcedo, Michelle P; Zon, Gerald; McCaffrey, Anton P; Paul, Natasha; Hogrefe, Richard I

2016-01-01

For most sample types, the automation of RNA and DNA sample preparation workflows enables high throughput next-generation sequencing (NGS) library preparation. Greater adoption of small RNA (sRNA) sequencing has been hindered by high sample input requirements and inherent ligation side products formed during library preparation. These side products, known as adapter dimer, are very similar in size to the tagged library. Most sRNA library preparation strategies thus employ a gel purification step to isolate tagged library from adapter dimer contaminants. At very low sample inputs, adapter dimer side products dominate the reaction and limit the sensitivity of this technique. Here we address the need for improved specificity of sRNA library preparation workflows with a novel library preparation approach that uses modified adapters to suppress adapter dimer formation. This workflow allows for lower sample inputs and elimination of the gel purification step, which in turn allows for an automatable sRNA library preparation protocol.

Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples.

PubMed

Quick, Joshua; Grubaugh, Nathan D; Pullan, Steven T; Claro, Ingra M; Smith, Andrew D; Gangavarapu, Karthik; Oliveira, Glenn; Robles-Sikisaka, Refugio; Rogers, Thomas F; Beutler, Nathan A; Burton, Dennis R; Lewis-Ximenez, Lia Laura; de Jesus, Jaqueline Goes; Giovanetti, Marta; Hill, Sarah C; Black, Allison; Bedford, Trevor; Carroll, Miles W; Nunes, Marcio; Alcantara, Luiz Carlos; Sabino, Ester C; Baylis, Sally A; Faria, Nuno R; Loose, Matthew; Simpson, Jared T; Pybus, Oliver G; Andersen, Kristian G; Loman, Nicholas J

2017-06-01

Genome sequencing has become a powerful tool for studying emerging infectious diseases; however, genome sequencing directly from clinical samples (i.e., without isolation and culture) remains challenging for viruses such as Zika, for which metagenomic sequencing methods may generate insufficient numbers of viral reads. Here we present a protocol for generating coding-sequence-complete genomes, comprising an online primer design tool, a novel multiplex PCR enrichment protocol, optimized library preparation methods for the portable MinION sequencer (Oxford Nanopore Technologies) and the Illumina range of instruments, and a bioinformatics pipeline for generating consensus sequences. The MinION protocol does not require an Internet connection for analysis, making it suitable for field applications with limited connectivity. Our method relies on multiplex PCR for targeted enrichment of viral genomes from samples containing as few as 50 genome copies per reaction. Viral consensus sequences can be achieved in 1-2 d by starting with clinical samples and following a simple laboratory workflow. This method has been successfully used by several groups studying Zika virus evolution and is facilitating an understanding of the spread of the virus in the Americas. The protocol can be used to sequence other viral genomes using the online Primal Scheme primer designer software. It is suitable for sequencing either RNA or DNA viruses in the field during outbreaks or as an inexpensive, convenient method for use in the lab.

Comparing dried and liquid blood serum samples of depressed patients: An analysis by Raman and infrared spectroscopy methods.

PubMed

Depciuch, J; Parlinska-Wojtan, M

2018-02-20

Depression is a serious mental illness. To study the mechanism of depression and search for new, more effective therapies, animal models are often used. Unfortunately, none of the available models reflects all the symptoms of depression. Therefore researchers are looking for new tools to diagnose depression. Unfortunately, the nowadays-available depression diagnosis methods are only psychological tests. However, it is known, that the amount of phospholipids, proteins and lipids decreases during depression. Raman and FTIR (Fourier Transform Infra Red) spectroscopies provide information on the chemical compounds in the measured sample e.g. blood serum. These spectroscopic techniques may thus become reliable and accurate tools for evaluating changes in the amount of phospholipids and proteins in depression disease. In this study differences between dried and liquid blood serum samples of healthy and depressed individuals measured by Raman (range 0-3000cm -1 ) and FTIR (Fourier Transform Infrared) (range 900-3000cm -1 ) spectroscopy were evaluated. The resulting spectra and accurate analysis led to the conclusion that an appropriate measurement of the background and the elimination of peaks from water had the greatest impact on the reliability of the results. Furthermore, after detailed studies of FTIR and Raman spectra of dried and liquid blood serum samples, including a complete analysis of peaks after Kramers-Kröning (KK) transformation, it was found that the sample preparation did not affect the results obtained by Raman spectroscopy. In FTIR measurements only a minimal effect on peak intensity was observed. Copyright © 2017 Elsevier B.V. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

This volume contains the interim change notice for sample preparation methods. Covered are: acid digestion for metals analysis, fusion of Hanford tank waste solids, water leach of sludges/soils/other solids, extraction procedure toxicity (simulate leach in landfill), sample preparation for gamma spectroscopy, acid digestion for radiochemical analysis, leach preparation of solids for free cyanide analysis, aqueous leach of solids for anion analysis, microwave digestion of glasses and slurries for ICP/MS, toxicity characteristic leaching extraction for inorganics, leach/dissolution of activated metal for radiochemical analysis, extraction of single-shell tank (SST) samples for semi-VOC analysis, preparation and cleanup of hydrocarbon- containing samples for VOCmore » and semi-VOC analysis, receiving of waste tank samples in onsite transfer cask, receipt and inspection of SST samples, receipt and extrusion of core samples at 325A shielded facility, cleaning and shipping of waste tank samplers, homogenization of solutions/slurries/sludges, and test sample preparation for bioassay quality control program.« less

Analytical tools for the analysis of β-carotene and its degradation products

PubMed Central

Stutz, H.; Bresgen, N.; Eckl, P. M.

2015-01-01

Abstract β-Carotene, the precursor of vitamin A, possesses pronounced radical scavenging properties. This has centered the attention on β-carotene dietary supplementation in healthcare as well as in the therapy of degenerative disorders and several cancer types. However, two intervention trials with β-carotene have revealed adverse effects on two proband groups, that is, cigarette smokers and asbestos-exposed workers. Beside other causative reasons, the detrimental effects observed have been related to the oxidation products of β-carotene. Their generation originates in the polyene structure of β-carotene that is beneficial for radical scavenging, but is also prone to oxidation. Depending on the dominant degradation mechanism, bond cleavage might occur either randomly or at defined positions of the conjugated electron system, resulting in a diversity of cleavage products (CPs). Due to their instability and hydrophobicity, the handling of standards and real samples containing β-carotene and related CPs requires preventive measures during specimen preparation, analyte extraction, and final analysis, to avoid artificial degradation and to preserve the initial analyte portfolio. This review critically discusses different preparation strategies of standards and treatment solutions, and also addresses their protection from oxidation. Additionally, in vitro oxidation strategies for the generation of oxidative model compounds are surveyed. Extraction methods are discussed for volatile and non-volatile CPs individually. Gas chromatography (GC), (ultra)high performance liquid chromatography (U)HPLC, and capillary electrochromatography (CEC) are reviewed as analytical tools for final analyte analysis. For identity confirmation of analytes, mass spectrometry (MS) is indispensable, and the appropriate ionization principles are comprehensively discussed. The final sections cover analysis of real samples and aspects of quality assurance, namely matrix effects and method validation. PMID:25867077

Recent mass spectrometry-based techniques and considerations for disulfide bond characterization in proteins.

PubMed

Lakbub, Jude C; Shipman, Joshua T; Desaire, Heather

2018-04-01

Disulfide bonds are important structural moieties of proteins: they ensure proper folding, provide stability, and ensure proper function. With the increasing use of proteins for biotherapeutics, particularly monoclonal antibodies, which are highly disulfide bonded, it is now important to confirm the correct disulfide bond connectivity and to verify the presence, or absence, of disulfide bond variants in the protein therapeutics. These studies help to ensure safety and efficacy. Hence, disulfide bonds are among the critical quality attributes of proteins that have to be monitored closely during the development of biotherapeutics. However, disulfide bond analysis is challenging because of the complexity of the biomolecules. Mass spectrometry (MS) has been the go-to analytical tool for the characterization of such complex biomolecules, and several methods have been reported to meet the challenging task of mapping disulfide bonds in proteins. In this review, we describe the relevant, recent MS-based techniques and provide important considerations needed for efficient disulfide bond analysis in proteins. The review focuses on methods for proper sample preparation, fragmentation techniques for disulfide bond analysis, recent disulfide bond mapping methods based on the fragmentation techniques, and automated algorithms designed for rapid analysis of disulfide bonds from liquid chromatography-MS/MS data. Researchers involved in method development for protein characterization can use the information herein to facilitate development of new MS-based methods for protein disulfide bond analysis. In addition, individuals characterizing biotherapeutics, especially by disulfide bond mapping in antibodies, can use this review to choose the best strategies for disulfide bond assignment of their biologic products. Graphical Abstract This review, describing characterization methods for disulfide bonds in proteins, focuses on three critical components: sample preparation, mass spectrometry data, and software tools.

The road map towards providing a robust Raman spectroscopy-based cancer diagnostic platform and integration into clinic

NASA Astrophysics Data System (ADS)

Lau, Katherine; Isabelle, Martin; Lloyd, Gavin R.; Old, Oliver; Shepherd, Neil; Bell, Ian M.; Dorney, Jennifer; Lewis, Aaran; Gaifulina, Riana; Rodriguez-Justo, Manuel; Kendall, Catherine; Stone, Nicolas; Thomas, Geraint; Reece, David

2016-03-01

Despite the demonstrated potential as an accurate cancer diagnostic tool, Raman spectroscopy (RS) is yet to be adopted by the clinic for histopathology reviews. The Stratified Medicine through Advanced Raman Technologies (SMART) consortium has begun to address some of the hurdles in its adoption for cancer diagnosis. These hurdles include awareness and acceptance of the technology, practicality of integration into the histopathology workflow, data reproducibility and availability of transferrable models. We have formed a consortium, in joint efforts, to develop optimised protocols for tissue sample preparation, data collection and analysis. These protocols will be supported by provision of suitable hardware and software tools to allow statistically sound classification models to be built and transferred for use on different systems. In addition, we are building a validated gastrointestinal (GI) cancers model, which can be trialled as part of the histopathology workflow at hospitals, and a classification tool. At the end of the project, we aim to deliver a robust Raman based diagnostic platform to enable clinical researchers to stage cancer, define tumour margin, build cancer diagnostic models and discover novel disease bio markers.

Structured illumination microscopy as a diagnostic tool for nephrotic disease

NASA Astrophysics Data System (ADS)

Nylk, Jonathan; Pullman, James M.; Campbell, Elaine C.; Gunn-Moore, Frank J.; Prystowsky, Michael B.; Dholakia, Kishan

2017-02-01

Nephrotic disease is a group of debilitating and sometimes lethal diseases affecting kidney function, specifically the loss of ability to retain vital proteins in the blood while smaller molecules are removed through filtration into the urine. Treatment routes are often dictated by microscopic analysis of kidney biopsies. Podocytes within the glomeruli of the kidney have many interdigitating projections (foot processes), which form the main filtration system. Nephrotic disease is characterised by the loss of this tightly interdigitating substructure and its observation by electron microscopy (EM) is necessitated as these structures are typically 250 500nm wide, with 40nm spacing. Diagnosis by EM is both expensive and time consuming; it can take up to one week to complete the preparation, imaging, and analysis of a single sample. We propose structured illumination microscopy (SIM) as an alternative, optical diagnostic tool. Our results show that SIM can resolve the structure of fluorescent probes tagged to podocin, a protein localised to the periphery of the podocyte foot processes. Three-dimensional podocin maps were acquired in healthy tissue and tissue from patients diagnosed with two different nephrotic disease states; minimal change disease and membranous nephropathy. These structures correlated well with EM images of the same structure. Preparation, imaging, and analysis could be achieved in several hours. Additionally, the volumetric information of the SIM images revealed morphological changes in disease states not observed by EM. This evidence supports the use of SIM as a diagnostic tool for nephrotic disease and can potentially reduce the time and cost per diagnosis.

Attenuated total reflectance spectroscopy of plant leaves: A tool for ecological and botanical studies

USGS Publications Warehouse

Ribeiro da Luz, B.

2006-01-01

??? Attenuated total reflectance (ATR) spectra of plant leaves display complex absorption features related to organic constituents of leaf surfaces. The spectra can be recorded rapidly, both in the field and in the laboratory, without special sample preparation. ??? This paper explores sources of ATR spectral variation in leaves, including compositional, positional and temporal variations. Interspecific variations are also examined, including the use of ATR spectra as a tool for species identification. ??? Positional spectral variations generally reflected the abundance of cutin and the epicuticular wax thickness and composition. For example, leaves exposed to full sunlight commonly showed more prominent cutin- and wax-related absorption features compared with shaded leaves. Adaxial vs. abaxial leaf surfaces displayed spectral variations reflecting differences in trichome abundance and wax composition. Mature vs. young leaves showed changes in absorption band position and intensity related to cutin, polysaccharide, and possibly amorphous silica development on and near the leaf surfaces. ??? Provided that similar samples are compared (e.g. adaxial surfaces of mature, sun-exposed leaves) same-species individuals display practically identical ATR spectra. Using spectral matching procedures to analyze an ATR database containing 117 individuals, including 32 different tree species, 83% of the individuals were correctly identified. ?? The Authors (2006).

EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

PubMed Central

Kalina, T; Flores-Montero, J; van der Velden, V H J; Martin-Ayuso, M; Böttcher, S; Ritgen, M; Almeida, J; Lhermitte, L; Asnafi, V; Mendonça, A; de Tute, R; Cullen, M; Sedek, L; Vidriales, M B; Pérez, J J; te Marvelde, J G; Mejstrikova, E; Hrusak, O; Szczepański, T; van Dongen, J J M; Orfao, A

2012-01-01

The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally comparable results among different laboratories. This required the selection of optimal combinations of compatible fluorochromes and the design and evaluation of adequate standard operating procedures (SOPs) for instrument setup, fluorescence compensation and sample preparation. Additionally, we developed software tools for the evaluation of individual antibody reagents and antibody panels. Each section describes what has been evaluated experimentally versus adopted based on existing data and experience. Multicentric evaluation demonstrated high levels of reproducibility based on strict implementation of the EuroFlow SOPs and antibody panels. Overall, the 6 years of extensive collaborative experiments and the analysis of hundreds of cell samples of patients and healthy controls in the EuroFlow centers have provided for the first time laboratory protocols and software tools for fully standardized 8-color flow cytometric immunophenotyping of normal and malignant leukocytes in bone marrow and blood; this has yielded highly comparable data sets, which can be integrated in a single database. PMID:22948490

Preparation of stable silica surfaces for surface forces measurement

NASA Astrophysics Data System (ADS)

Ren, Huai-Yin; Mizukami, Masashi; Kurihara, Kazue

2017-09-01

A surface forces apparatus (SFA) measures the forces between two surfaces as a function of the surface separation distance. It is regarded as an essential tool for studying the interactions between two surfaces. However, sample surfaces used for the conventional SFA measurements have been mostly limited to thin (ca. 2-3 μm) micas, which are coated with silver layers (ca. 50 nm) on their back, due to the requirement of the distance determination by transmission mode optical interferometry called FECO (fringes of equal chromatic order). The FECO method has the advantage of determining the absolute distance, so it should be important to increase the availability of samples other than mica, which is chemically nonreactive and also requires significant efforts for cleaving. Recently, silica sheets have been occasionally used in place of mica, which increases the possibility of surface modification. However, in this case, the silver layer side of the sheet is glued on a cylindrical quartz disc using epoxy resin, which is not stable in organic solvents and can be easily swollen or dissolved. The preparation of substrates more stable under severe conditions, such as in organic solvents, is necessary for extending application of the measurement. In this study, we report an easy method for preparing stable silica layers of ca. 2 μm in thickness deposited on gold layers (41 nm)/silica discs by sputtering, then annealed to enhance the stability. The obtained silica layers were stable and showed no swelling in organic solvents such as ethanol and toluene.

Potential artifacts associated with historical preparation of joint compound samples and reported airborne asbestos concentrations.

PubMed

Brorby, G P; Sheehan, P J; Berman, D W; Bogen, K T; Holm, S E

2011-05-01

Airborne samples collected in the 1970s for drywall workers using asbestos-containing joint compounds were likely prepared and analyzed according to National Institute of Occupational Safety and Health Method P&CAM 239, the historical precursor to current Method 7400. Experimentation with a re-created, chrysotile-containing, carbonate-based joint compound suggested that analysis following sample preparation by the historical vs. current method produces different fiber counts, likely because of an interaction between the different clearing and mounting chemicals used and the carbonate-based joint compound matrix. Differences were also observed during analysis using Method 7402, depending on whether acetic acid/dimethylformamide or acetone was used during preparation to collapse the filter. Specifically, air samples of sanded chrysotile-containing joint compound prepared by the historical method yielded fiber counts significantly greater (average of 1.7-fold, 95% confidence interval: 1.5- to 2.0-fold) than those obtained by the current method. In addition, air samples prepared by Method 7402 using acetic acid/dimethylformamide yielded fiber counts that were greater (2.8-fold, 95% confidence interval: 2.5- to 3.2-fold) than those prepared by this method using acetone. These results indicated (1) there is an interaction between Method P&CAM 239 preparation chemicals and the carbonate-based joint compound matrix that reveals fibers that were previously bound in the matrix, and (2) the same appeared to be true for Method 7402 preparation chemicals acetic acid/dimethylformamide. This difference in fiber counts is the opposite of what has been reported historically for samples of relatively pure chrysotile dusts prepared using the same chemicals. This preparation artifact should be considered when interpreting historical air samples for drywall workers prepared by Method P&CAM 239. Copyright © 2011 JOEH, LLC

Sample Preparation of Corn Seed Tissue to Prevent Analyte Relocations for Mass Spectrometry Imaging

NASA Astrophysics Data System (ADS)

Kim, Shin Hye; Kim, Jeongkwon; Lee, Young Jin; Lee, Tae Geol; Yoon, Sohee

2017-08-01

Corn seed tissue sections were prepared by the tape support method using an adhesive tape, and mass spectrometry imaging (MSI) was performed. The effect of heat generated during sample preparation was investigated by time-of-flight secondary mass spectrometry (TOF-SIMS) imaging of corn seed tissue prepared by the tape support and the thaw-mounted methods. Unlike thaw-mounted sample preparation, the tape support method does not cause imaging distortion because of the absence of heat, which can cause migration of the analytes on the sample. By applying the tape-support method, the corn seed tissue was prepared without structural damage and MSI with accurate spatial information of analytes was successfully performed.

MARS: bringing the automation of small-molecule bioanalytical sample preparations to a new frontier.

PubMed

Li, Ming; Chou, Judy; Jing, Jing; Xu, Hui; Costa, Aldo; Caputo, Robin; Mikkilineni, Rajesh; Flannelly-King, Shane; Rohde, Ellen; Gan, Lawrence; Klunk, Lewis; Yang, Liyu

2012-06-01

In recent years, there has been a growing interest in automating small-molecule bioanalytical sample preparations specifically using the Hamilton MicroLab(®) STAR liquid-handling platform. In the most extensive work reported thus far, multiple small-molecule sample preparation assay types (protein precipitation extraction, SPE and liquid-liquid extraction) have been integrated into a suite that is composed of graphical user interfaces and Hamilton scripts. Using that suite, bioanalytical scientists have been able to automate various sample preparation methods to a great extent. However, there are still areas that could benefit from further automation, specifically, the full integration of analytical standard and QC sample preparation with study sample extraction in one continuous run, real-time 2D barcode scanning on the Hamilton deck and direct Laboratory Information Management System database connectivity. We developed a new small-molecule sample-preparation automation system that improves in all of the aforementioned areas. The improved system presented herein further streamlines the bioanalytical workflow, simplifies batch run design, reduces analyst intervention and eliminates sample-handling error.

An optimised protocol for molecular identification of Eimeria from chickens.

PubMed

Kumar, Saroj; Garg, Rajat; Moftah, Abdalgader; Clark, Emily L; Macdonald, Sarah E; Chaudhry, Abdul S; Sparagano, Olivier; Banerjee, Partha S; Kundu, Krishnendu; Tomley, Fiona M; Blake, Damer P

2014-01-17

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples. Copyright © 2013 Dirk Vulpius The Authors. Published by Elsevier B.V. All rights reserved.

Online Student Collaboration and FERPA Considerations

ERIC Educational Resources Information Center

Schrameyer, Alexander R.; Graves, Tracy M.; Hua, David M.; Brandt, Nile C.

2016-01-01

The increasing use of online or cloud collaboration tools in organizations creates a need for students to develop skills in the use of these tools. Instructors have an opportunity to prepare students by incorporating these collaboration tools into their courses. The use of these online collaboration tools has the potential to expose instructors…

Finite element simulation of cutting grey iron HT250 by self-prepared Si3N4 ceramic insert

NASA Astrophysics Data System (ADS)

Wang, Bo; Wang, Li; Zhang, Enguang

2017-04-01

The finite element method has been able to simulate and solve practical machining problems, achieve the required accuracy and the highly reliability. In this paper, the simulation models based on the material properties of the self-prepared Si3N4 insert and HT250 were created. Using these models, the results of cutting force, cutting temperature and tool wear rate were obtained, and tool wear mode was predicted after cutting simulation. These approaches may develop as the new method for testing new cutting-tool materials, shortening development cycle and reducing the cost.

Analytical aspects of hydrogen exchange mass spectrometry

PubMed Central

Engen, John R.; Wales, Thomas E.

2016-01-01

The analytical aspects of measuring hydrogen exchange by mass spectrometry are reviewed. The nature of analytical selectivity in hydrogen exchange is described followed by review of the analytical tools required to accomplish fragmentation, separation, and the mass spectrometry measurements under restrictive exchange quench conditions. In contrast to analytical quantitation that relies on measurements of peak intensity or area, quantitation in hydrogen exchange mass spectrometry depends on measuring a mass change with respect to an undeuterated or deuterated control, resulting in a value between zero and the maximum amount of deuterium that could be incorporated. Reliable quantitation is a function of experimental fidelity and to achieve high measurement reproducibility, a large number of experimental variables must be controlled during sample preparation and analysis. The method also reports on important qualitative aspects of the sample, including conformational heterogeneity and population dynamics. PMID:26048552

Cross-sectional TEM specimen preparation for W/B{sub 4}C multilayer sample using FIB

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mondal, Puspen, E-mail: puspen@rrcat.gov.in; Pradhan, P. C.; Tiwari, Pragya

2016-05-23

A recent emergence of a cross-beam scanning electron microscopy (SEM)/focused-ion-beam (FIB) system have given choice to fabricate cross-sectional transmission electron microscopy (TEM) specimen of thin film multilayer sample. A 300 layer pair thin film multilayer sample of W/B{sub 4}C was used to demonstrate the specimen lift-out technique in very short time as compared to conventional cross-sectional sample preparation technique. To get large area electron transparent sample, sample prepared by FIB is followed by Ar{sup +} ion polishing at 2 kV with grazing incident. The prepared cross-sectional sample was characterized by transmission electron microscope.

Developing and Evaluating Multimedia Patient Education Tools to Better Prepare Prostate-Cancer Patients for Radiotherapy Treatment (Randomized Study).

PubMed

Dawdy, Krista; Bonin, Katija; Russell, Steve; Ryzynski, Agnes; Harth, Tamara; Townsend, Christopher; Liu, Stanley; Chu, William; Cheung, Patrick; Chung, Hans; Morton, Gerard; Vesprini, Danny; Loblaw, Andrew; Cao, Xingshan; Szumacher, Ewa

2018-06-01

The purpose of this study is to determine the effectiveness of multimedia educational tools to improve CT planning preparation for intensity modulated radiotherapy (IMRT) for prostate cancer. Many patients are not prepared when given verbal preparation instructions to have a full bladder and empty rectum for their IMRT and require being rescanned, which results in additional costs for the patient and the hospital. A pamphlet and video outlining the proper preparation for prostate IMRT was created to decrease additional scans and the associated costs, while increasing patient satisfaction. A controlled, randomized experimental group study was conducted to examine the effectiveness of the multimedia tools (the video and the pamphlet), as compared to the pamphlet only, in preparing patients for their planning CT appointment. We found no statistical difference between the multimedia group and the pamphlet group in patients' preparedness for their appointments and the rescanning rate. However, patients in the multimedia group indicated that they felt more prepared about their treatment after watching the video and stated that they would recommend the video to other patients with prostate cancer. Furthermore, patients who had to wait longer for their planning CT appointment felt less prepared by the materials than those with a shorter wait time. We recommend reducing wait times between appointments as much as possible to increase patients' preparedness for the planning CT. We conclude that providing multimedia treatment information and minimizing wait times increases patients' feelings of preparedness leading to a more positive treatment experience and reducing costly rescans. ClinicalTrials.gov NCT02410291.

Improving Store Separation Six-Degree-of-Freedom Tools (ISSSDF)

DTIC Science & Technology

2007-01-01

the Australian Air Force. NAVAIR is in the process of preparing a flight clearance for the High Speed Anti- radiation Demonstrator ( HSAD ...program. This clearance will be prepared using the conventional tool NAVSEP. The HSAD missile, however, utilizes an autopilot to move four movable...conventional NAVSEP approach to clear the HSAD missile, it would be desirable to test the TGP and STEME codes for the same case. It would be

System for autonomous monitoring of bioagents

DOEpatents

Langlois, Richard G.; Milanovich, Fred P.; Colston, Jr, Billy W.; Brown, Steve B.; Masquelier, Don A.; Mariella, Jr., Raymond P.; Venkateswaran, Kodomudi

2015-06-09

An autonomous monitoring system for monitoring for bioagents. A collector gathers the air, water, soil, or substance being monitored. A sample preparation means for preparing a sample is operatively connected to the collector. A detector for detecting the bioagents in the sample is operatively connected to the sample preparation means. One embodiment of the present invention includes confirmation means for confirming the bioagents in the sample.

Concept Maps: A Tool to Prepare for High Fidelity Simulation in Nursing

ERIC Educational Resources Information Center

Daley, Barbara J.; Beman, Sarah Black; Morgan, Sarah; Kennedy, Linda; Sheriff, Mandy

2017-01-01

In this study, the use of concept mapping as a method to prepare for high fidelity simulated learning experiences was investigated. Fourth year baccalaureate nursing students were taught how to use concept maps as a way to prepare for high fidelity simulated nursing experiences. Students prepared concept maps for two simulated experiences…

Comparative Characterization of Crofelemer Samples Using Data Mining and Machine Learning Approaches With Analytical Stability Data Sets.

PubMed

Nariya, Maulik K; Kim, Jae Hyun; Xiong, Jian; Kleindl, Peter A; Hewarathna, Asha; Fisher, Adam C; Joshi, Sangeeta B; Schöneich, Christian; Forrest, M Laird; Middaugh, C Russell; Volkin, David B; Deeds, Eric J

2017-11-01

There is growing interest in generating physicochemical and biological analytical data sets to compare complex mixture drugs, for example, products from different manufacturers. In this work, we compare various crofelemer samples prepared from a single lot by filtration with varying molecular weight cutoffs combined with incubation for different times at different temperatures. The 2 preceding articles describe experimental data sets generated from analytical characterization of fractionated and degraded crofelemer samples. In this work, we use data mining techniques such as principal component analysis and mutual information scores to help visualize the data and determine discriminatory regions within these large data sets. The mutual information score identifies chemical signatures that differentiate crofelemer samples. These signatures, in many cases, would likely be missed by traditional data analysis tools. We also found that supervised learning classifiers robustly discriminate samples with around 99% classification accuracy, indicating that mathematical models of these physicochemical data sets are capable of identifying even subtle differences in crofelemer samples. Data mining and machine learning techniques can thus identify fingerprint-type attributes of complex mixture drugs that may be used for comparative characterization of products. Copyright © 2017 American Pharmacists Association®. All rights reserved.

Laboratory preparation questionnaires as a tool for the implementation of the Just in Time Teaching in the Physics I laboratories: Research training

NASA Astrophysics Data System (ADS)

Miranda, David A.; Sanchez, Melba J.; Forero, Oscar M.

2017-06-01

The implementation of the JiTT (Just in Time Teaching) strategy is presented to increase the previous preparation of students enrolled in the subject Physics Laboratory I offered at the Industrial University of Santander (UIS), Colombia. In this study, a laboratory preparation questionnaire (CPL) was applied as a tool for the implementation of JiTT combined with elements of mediated learning. It was found that the CPL allows to improve the students’ experience regarding the preparation of the laboratory and the development of the experimental session. These questionnaires were implemented in an academic manager (Moodle) and a web application (lab.ciencias.uis.edu.co) was used to publish the contents essential for the preparation of the student before each practical session. The most significant result was that the students performed the experimental session with the basic knowledge to improve their learning experience.

Method development towards qualitative and semi-quantitative analysis of multiple pesticides from food surfaces and extracts by desorption electrospray ionization mass spectrometry as a preselective tool for food control.

PubMed

Gerbig, Stefanie; Stern, Gerold; Brunn, Hubertus E; Düring, Rolf-Alexander; Spengler, Bernhard; Schulz, Sabine

2017-03-01

Direct analysis of fruit and vegetable surfaces is an important tool for in situ detection of food contaminants such as pesticides. We tested three different ways to prepare samples for the qualitative desorption electrospray ionization mass spectrometry (DESI-MS) analysis of 32 pesticides found on nine authentic fruits collected from food control. Best recovery rates for topically applied pesticides (88%) were found by analyzing the surface of a glass slide which had been rubbed against the surface of the food. Pesticide concentration in all samples was at or below the maximum residue level allowed. In addition to the high sensitivity of the method for qualitative analysis, quantitative or, at least, semi-quantitative information is needed in food control. We developed a DESI-MS method for the simultaneous determination of linear calibration curves of multiple pesticides of the same chemical class using normalization to one internal standard (ISTD). The method was first optimized for food extracts and subsequently evaluated for the quantification of pesticides in three authentic food extracts. Next, pesticides and the ISTD were applied directly onto food surfaces, and the corresponding calibration curves were obtained. The determination of linear calibration curves was still feasible, as demonstrated for three different food surfaces. This proof-of-principle method was used to simultaneously quantify two pesticides on an authentic sample, showing that the method developed could serve as a fast and simple preselective tool for disclosure of pesticide regulation violations. Graphical Abstract Multiple pesticide residues were detected and quantified in-situ from an authentic set of food items and extracts in a proof of principle study.

Spatially resolved chemical analysis of cicada wings using laser-ablation electrospray ionization (LAESI) imaging mass spectrometry (IMS).

PubMed

Román, Jessica K; Walsh, Callee M; Oh, Junho; Dana, Catherine E; Hong, Sungmin; Jo, Kyoo D; Alleyne, Marianne; Miljkovic, Nenad; Cropek, Donald M

2018-03-01

Laser-ablation electrospray ionization (LAESI) imaging mass spectrometry (IMS) is an emerging bioanalytical tool for direct imaging and analysis of biological tissues. Performing ionization in an ambient environment, this technique requires little sample preparation and no additional matrix, and can be performed on natural, uneven surfaces. When combined with optical microscopy, the investigation of biological samples by LAESI allows for spatially resolved compositional analysis. We demonstrate here the applicability of LAESI-IMS for the chemical analysis of thin, desiccated biological samples, specifically Neotibicen pruinosus cicada wings. Positive-ion LAESI-IMS accurate ion-map data was acquired from several wing cells and superimposed onto optical images allowing for compositional comparisons across areas of the wing. Various putative chemical identifications were made indicating the presence of hydrocarbons, lipids/esters, amines/amides, and sulfonated/phosphorylated compounds. With the spatial resolution capability, surprising chemical distribution patterns were observed across the cicada wing, which may assist in correlating trends in surface properties with chemical distribution. Observed ions were either (1) equally dispersed across the wing, (2) more concentrated closer to the body of the insect (proximal end), or (3) more concentrated toward the tip of the wing (distal end). These findings demonstrate LAESI-IMS as a tool for the acquisition of spatially resolved chemical information from fragile, dried insect wings. This LAESI-IMS technique has important implications for the study of functional biomaterials, where understanding the correlation between chemical composition, physical structure, and biological function is critical. Graphical abstract Positive-ion laser-ablation electrospray ionization mass spectrometry coupled with optical imaging provides a powerful tool for the spatially resolved chemical analysis of cicada wings.

Use of Malachite Green-Loop Mediated Isothermal Amplification for Detection of Plasmodium spp. Parasites

PubMed Central

Lucchi, Naomi W.; Ljolje, Dragan; Silva-Flannery, Luciana; Udhayakumar, Venkatachalam

2016-01-01

Malaria elimination efforts are hampered by the lack of sensitive tools to detect infections with low-level parasitemia, usually below the threshold of standard diagnostic methods, microscopy and rapid diagnostic tests. Isothermal nucleic acid amplification assays such as the loop-mediated isothermal amplification (LAMP), are well suited for field use as they do not require thermal cyclers to run the test. However, the use of specialized equipment, as described by many groups, reduces the versatility of the LAMP technique as a simple tool for use in endemic countries. In this study, the use of the malachite green (MG) dye, as a visual endpoint readout, together with a simple mini heat block was evaluated for the detection of malaria parasites. The assay was performed for 1 hour at 63°C and the results scored by 3 independent human readers. The limit of detection of the assay was determined using well-quantified Plasmodium spp. infected reference samples and its utility in testing clinical samples was determined using 190 pre-treatment specimens submitted for reference diagnosis of imported malaria in the United States. Use of a simplified boil and spin methods of DNA extraction from whole blood and filter paper was also investigated. We demonstrate the accurate and sensitive detection of malaria parasites using this assay with a detection limit ranging between 1–8 parasites/μL, supporting its applicability for the detection of infections with low parasite burden. This assay is compatible with the use of a simple boil and spin sample preparation method from both whole blood and filter papers without a loss of sensitivity. The MG-LAMP assay described here has great potential to extend the reach of molecular tools to settings where they are needed. PMID:26967908

Extemporaneously preparative biodegradable injectable polymer systems exhibiting temperature-responsive irreversible gelation.

PubMed

Yoshida, Yasuyuki; Takata, Kazuyuki; Takai, Hiroki; Kawahara, Keisuke; Kuzuya, Akinori; Ohya, Yuichi

2017-10-01

On clinical application of biodegradable injectable polymer (IP) systems, quick extemporaneous preparation of IP formulations and longer duration time gel state after injection into the body are the important targets to be developed. Previously, we had reported temperature-responsive covalent gelation systems via bio-orthogonal thiol-ene reaction by 'mixing strategy' of amphiphilic biodegradable tri-block copolymer (tri-PCG) attaching acryloyl groups on both termini (tri-PCG-Acryl) with reactive polythiol. In other previous works, we found 'freeze-dry with PEG/dispersion' method as quick extemporaneous preparation method of biodegradable IP formulations. In this study, we applied this quick preparative method to the temperature-triggered covalent gelation system. The instant formulation (D-sample) could be prepared by 'freeze-dry with PEG/dispersion' just mixing of tri-PCG-Acryl micelle dispersion and tri-PCG/DPMP micelle dispersion with PEG, that can be prepared in 30 s from the dried samples. The obtained D-sample showed irreversible gelation and long duration time of gel state, which was basically the same as the formulations prepared by the usual heating dissolution method (S-sample). Interestingly, the D-sample could maintain its sol state for a longer time (24 h) after preparing the formulation at r.t. compared with the S-sample, which became a gel in 3 h after preparing. The IP system showed good biocompatibility and long duration time of the gel state after subcutaneous implantation. These characteristics of D-samples, quick extemporaneous preparation and high stability in the sol state before injection, would be very convenient in a clinical setting.

Effect of Genetic Database Comprehensiveness on Fractional Proteomics of Escherichia coli O157:H7

DTIC Science & Technology

2014-01-01

proteins would be observed in the extracellular fraction. 15. SUBJECT TERMS Escherichia coli O157:H7 Liquid chromatography Mass spectrometry...Preparation ...............1 2.2 Liquid Chromatography /Mass Spectrometry Sample Preparation ....................2 2.3 Liquid Chromatography /Mass... Chromatography /Mass Spectrometry Sample Preparation. Samples were prepared for liquid chromatography tandem mass spectrometry (LC-MS/MS) in a similar

Utility of Cytospin and Cell block Technology in Evaluation of Body Fluids and Urine Samples: A Comparative Study.

PubMed

Qamar, Irmeen; Rehman, Suhailur; Mehdi, Ghazala; Maheshwari, Veena; Ansari, Hena A; Chauhan, Sunanda

2018-01-01

Cytologic examination of body fluids commonly involves the use of direct or sediment smears, cytocentrifuge preparations, membrane filter preparations, or cell block sections. Cytospin and cell block techniques are extremely useful in improving cell yield of thin serous effusions and urine samples, and ensure high diagnostic efficacy. We studied cytospin preparations and cell block sections prepared from 180 samples of body fluids and urine samples to compare the relative efficiency of cell retrieval, preservation of cell morphology, ease of application of special stains, and diagnostic efficacy. Samples were collected and processed to prepare cytospin smears and cell block sections. We observed that overall, cell yield and preservation of individual cell morphology were better in cytospin preparations as compared to cell blocks, while preservation of architectural pattern was better in cell block sections. The number of suspicious cases also decreased on cell block sections, with increased detection of malignancy. It was difficult to prepare cell blocks from urine samples due to low cellularity. Cytospin technology is a quick, efficient, and cost-effective method of increasing cell yield in hypocellular samples, with better preservation of cell morphology. Cell blocks are better prepared from high cellularity fluids; however, tissue architecture is better studied, with improved rate of diagnosis and decrease in ambiguous results. Numerous sections can be prepared from a small amount of material. Special stains and immunochemical stains can be easily applied to cell blocks. It also provides a source of archival material.

Computer program for determining rotational line intensity factors for diatomic molecules

NASA Technical Reports Server (NTRS)

Whiting, E. E.

1973-01-01

A FORTRAN IV computer program, that provides a new research tool for determining reliable rotational line intensity factors (also known as Honl-London factors), for most electric and magnetic dipole allowed diatomic transitions, is described in detail. This users manual includes instructions for preparing the input data, a program listing, detailed flow charts, and three sample cases. The program is applicable to spin-allowed dipole transitions with either or both states intermediate between Hund's case (a) and Hund's case (b) coupling and to spin-forbidden dipole transitions with either or both states intermediate between Hund's case (c) and Hund's case (b) coupling.

Sugar microanalysis by HPLC with benzoylation: improvement via introduction of a C-8 cartridge and a high efficiency ODS column.

PubMed

Miyagi, Michiko; Yokoyama, Hirokazu; Hibi, Toshifumi

2007-07-01

An HPLC protocol for sugar microanalysis based on the formation of ultraviolet-absorbing benzoyl chloride derivatives was improved. Here, samples were prepared with a C-8 cartridge and analyzed with a high efficiency ODS column, in which porous spherical silica particles 3 microm in diameter were packed. These devices allowed us to simultaneously quantify multiple sugars and sugar alcohols up to 10 ng/ml and to provide satisfactory separations of some sugars, such as fructose and myo-inositol and sorbitol and mannitol. This protocol, which does not require special apparatuses, should become a powerful tool in sugar research.

Filter paper-assisted cell transfer (FaCT) technique: A novel cell-sampling technique for intraoperative diagnosis of central nervous system tumors.

PubMed

Kawamura, Jumpei; Kamoshida, Shingo; Shimakata, Takaaki; Hayashi, Yurie; Sakamaki, Kuniko; Denda, Tamami; Kawai, Kenji; Kuwao, Sadahito

2017-04-01

Intraoperative diagnosis of central nervous system (CNS) tumors provides critical guidance to surgeons in the determination of surgical resection margins and treatment. The techniques and preparations used for the intraoperative diagnosis of CNS tumors include frozen sectioning and cytologic methods (squash smear and touch imprint). Cytologic specimens, which do not have freezing artifacts, are important as an adjuvant tool to frozen sections. However, if the amount of submitted tissue samples is limited, then it is difficult to prepare both frozen sections and squash smears or touch imprint specimens from a single sample at the same time. Therefore, the objective of this study was to derive cells directly from filter paper on which tumor samples are placed. The authors established the filter paper-assisted cell transfer (FaCT) smear technique, in which tumor cells are transferred onto a glass slide directly from the filter paper sample spot after the biopsy is removed. Cell yields and diagnostic accuracy of the FaCT smears were assessed in 40 CNS tumors. FaCT smears had ample cell numbers and well preserved cell morphology sufficient for cytologic diagnosis, even if the submitted tissues were minimal. The overall diagnostic concordance rates between frozen sections and FaCT smears were 90% and 87.5%, respectively (no significant differences). When combining FaCT smears with frozen sections, the diagnostic concordance rate rose to 92.5%. The current results suggest that the FaCT smear technique is a simple and effective processing method that has significant value for intraoperative diagnosis of CNS tumors. Cancer Cytopathol 2017;125:277-282. © 2016 American Cancer Society. © 2017 American Cancer Society.

Influence of sample preparation and reliability of automated numerical refocusing in stain-free analysis of dissected tissues with quantitative phase digital holographic microscopy

NASA Astrophysics Data System (ADS)

Kemper, Björn; Lenz, Philipp; Bettenworth, Dominik; Krausewitz, Philipp; Domagk, Dirk; Ketelhut, Steffi

2015-05-01

Digital holographic microscopy (DHM) has been demonstrated to be a versatile tool for high resolution non-destructive quantitative phase imaging of surfaces and multi-modal minimally-invasive monitoring of living cell cultures in-vitro. DHM provides quantitative monitoring of physiological processes through functional imaging and structural analysis which, for example, gives new insight into signalling of cellular water permeability and cell morphology changes due to toxins and infections. Also the analysis of dissected tissues quantitative DHM phase contrast prospects application fields by stain-free imaging and the quantification of tissue density changes. We show that DHM allows imaging of different tissue layers with high contrast in unstained tissue sections. As the investigation of fixed samples represents a very important application field in pathology, we also analyzed the influence of the sample preparation. The retrieved data demonstrate that the quality of quantitative DHM phase images of dissected tissues depends strongly on the fixing method and common staining agents. As in DHM the reconstruction is performed numerically, multi-focus imaging is achieved from a single digital hologram. Thus, we evaluated the automated refocussing feature of DHM for application on different types of dissected tissues and revealed that on moderately stained samples highly reproducible holographic autofocussing can be achieved. Finally, it is demonstrated that alterations of the spatial refractive index distribution in murine and human tissue samples represent a reliable absolute parameter that is related of different degrees of inflammation in experimental colitis and Crohn's disease. This paves the way towards the usage of DHM in digital pathology for automated histological examinations and further studies to elucidate the translational potential of quantitative phase microscopy for the clinical management of patients, e.g., with inflammatory bowel disease.

Experimental Evaluation of the Tools of the Mind Pre-K Curriculum. Technical Report. Working Paper

ERIC Educational Resources Information Center

Farran, Dale C.; Wilson, Sandra J.; Meador, Deanna; Norvell, Jennifer; Nesbitt, Kimberly

2015-01-01

The experimental evaluation of the "Tools of the Mind Pre-K Curriculum" described in this report was designed to examine the effectiveness of the "Tools of the Mind" ("Tools") curriculum for enhancing children's self-regulation skills and their academic preparation for kindergarten when compared to the usual…

Social Networking Tools and Teacher Education Learning Communities: A Case Study

ERIC Educational Resources Information Center

Poulin, Michael T.

2014-01-01

Social networking tools have become an integral part of a pre-service teacher's educational experience. As a result, the educational value of social networking tools in teacher preparation programs must be examined. The specific problem addressed in this study is that the role of social networking tools in teacher education learning communities…

Evaluation of Training Samples Manually Welded With the Universal Handtool in a Space Simulation Chamber

NASA Technical Reports Server (NTRS)

Russell, C. K.; Malone, T. W.; Cato, S. N.

2004-01-01

The international space welding experiment was designed to evaluate the universal handtool (UHT) functions as a welding, brazing, coating, and cutting tool for in-space operations. The UHT is an electron beam welding system developed by the Paton Welding Institute (PWI), Kiev, Ukraine, and operated a 8 kV with up to 1 kW of power. In preparation for conducting the space welding experiment, cosmonauts were trained to properly operate the UHT and correctly process samples. This Technical Memorandum presents the results of the destructive and nondestructive evaluation of the training samples made in Russia in 1998. It was concluded that acceptable welds can be made with the UHT despite the constraints imposed by a space suit. The lap joint fillet weld configuration was more suitable than the butt joint configuration for operators with limited welding experience. The tube braze joint configuration designed by the PWI was easily brazed in a repeatable manner.

Sample Preparation for Electron Probe Microanalysis—Pushing the Limits

PubMed Central

Geller, Joseph D.; Engle, Paul D.

2002-01-01

There are two fundamental considerations in preparing samples for electron probe microanalysis (EPMA). The first one may seem obvious, but we often find it is overlooked. That is, the sample analyzed should be representative of the population from which it comes. The second is a direct result of the assumptions in the calculations used to convert x-ray intensity ratios, between the sample and standard, to concentrations. Samples originate from a wide range of sources. During their journey to being excited under the electron beam for the production of x rays there are many possibilities for sample alteration. Handling can contaminate samples by adding extraneous matter. In preparation, the various abrasives used in sizing the sample by sawing, grinding and polishing can embed themselves. The most accurate composition of a contaminated sample is, at best, not representative of the original sample; it is misleading. Our laboratory performs EPMA analysis on customer submitted samples and prepares over 250 different calibration standards including pure elements, compounds, alloys, glasses and minerals. This large variety of samples does not lend itself to mass production techniques, including automatic polishing. Our manual preparation techniques are designed individually for each sample. The use of automated preparation equipment does not lend itself to this environment, and is not included in this manuscript. The final step in quantitative electron probe microanalysis is the conversion of x-ray intensities ratios, known as the “k-ratios,” to composition (in mass fraction or atomic percent) and/or film thickness. Of the many assumptions made in the ZAF (where these letters stand for atomic number, absorption and fluorescence) corrections the localized geometry between the sample and electron beam, or takeoff angle, must be accurately known. Small angular errors can lead to significant errors in the final results. The sample preparation technique then becomes very important, and, under certain conditions, may even be the limiting factor in the analytical uncertainty budget. This paper considers preparing samples to get known geometries. It will not address the analysis of samples with irregular, unprepared surfaces or unknown geometries. PMID:27446757

A Systematic Review of Reporting Tools Applicable to Sexual and Reproductive Health Programmes: Step 1 in Developing Programme Reporting Standards

PubMed Central

Ali, Moazzam; Chandra-Mouli, Venkatraman; Tran, Nhan; Gülmezoglu, A. Metin

2015-01-01

Background Complete and accurate reporting of programme preparation, implementation and evaluation processes in the field of sexual and reproductive health (SRH) is essential to understand the impact of SRH programmes, as well as to guide their replication and scale-up. Objectives To provide an overview of existing reporting tools and identify core items used in programme reporting with a focus on programme preparation, implementation and evaluation processes. Methods A systematic review was completed for the period 2000–2014. Reporting guidelines, checklists and tools, irrespective of study design, applicable for reporting on programmes targeting SRH outcomes, were included. Two independent reviewers screened the title and abstract of all records. Full texts were assessed in duplicate, followed by data extraction on the focus, content area, year of publication, validation and description of reporting items. Data was synthesized using an iterative thematic approach, where items related to programme preparation, implementation and evaluation in each tool were extracted and aggregated into a consolidated list. Results Out of the 3,656 records screened for title and abstracts, full texts were retrieved for 182 articles, out of which 108 were excluded. Seventy-four full text articles corresponding to 45 reporting tools were retained for synthesis. The majority of tools were developed for reporting on intervention research (n = 15), randomized controlled trials (n = 8) and systematic reviews (n = 7). We identified a total of 50 reporting items, across three main domains and corresponding sub-domains: programme preparation (objective/focus, design, piloting); programme implementation (content, timing/duration/location, providers/staff, participants, delivery, implementation outcomes), and programme evaluation (process evaluation, implementation barriers/facilitators, outcome/impact evaluation). Conclusions Over the past decade a wide range of tools have been developed to improve the reporting of health research. Development of Programme Reporting Standards (PRS) for SRH can fill a significant gap in existing reporting tools. This systematic review is the first step in the development of such standards. In the next steps, we will draft a preliminary version of the PRS based on the aggregate list of identified items, and finalize the tool using a consensus process among experts and user-testing. PMID:26418859

A Systematic Review of Reporting Tools Applicable to Sexual and Reproductive Health Programmes: Step 1 in Developing Programme Reporting Standards.

PubMed

Kågesten, Anna; Tunçalp, Ӧzge; Ali, Moazzam; Chandra-Mouli, Venkatraman; Tran, Nhan; Gülmezoglu, A Metin

2015-01-01

Complete and accurate reporting of programme preparation, implementation and evaluation processes in the field of sexual and reproductive health (SRH) is essential to understand the impact of SRH programmes, as well as to guide their replication and scale-up. To provide an overview of existing reporting tools and identify core items used in programme reporting with a focus on programme preparation, implementation and evaluation processes. A systematic review was completed for the period 2000-2014. Reporting guidelines, checklists and tools, irrespective of study design, applicable for reporting on programmes targeting SRH outcomes, were included. Two independent reviewers screened the title and abstract of all records. Full texts were assessed in duplicate, followed by data extraction on the focus, content area, year of publication, validation and description of reporting items. Data was synthesized using an iterative thematic approach, where items related to programme preparation, implementation and evaluation in each tool were extracted and aggregated into a consolidated list. Out of the 3,656 records screened for title and abstracts, full texts were retrieved for 182 articles, out of which 108 were excluded. Seventy-four full text articles corresponding to 45 reporting tools were retained for synthesis. The majority of tools were developed for reporting on intervention research (n = 15), randomized controlled trials (n = 8) and systematic reviews (n = 7). We identified a total of 50 reporting items, across three main domains and corresponding sub-domains: programme preparation (objective/focus, design, piloting); programme implementation (content, timing/duration/location, providers/staff, participants, delivery, implementation outcomes), and programme evaluation (process evaluation, implementation barriers/facilitators, outcome/impact evaluation). Over the past decade a wide range of tools have been developed to improve the reporting of health research. Development of Programme Reporting Standards (PRS) for SRH can fill a significant gap in existing reporting tools. This systematic review is the first step in the development of such standards. In the next steps, we will draft a preliminary version of the PRS based on the aggregate list of identified items, and finalize the tool using a consensus process among experts and user-testing.

Assessment of the effects of different sample perfusion procedures on phase-contrast tomographic images of mouse spinal cord

NASA Astrophysics Data System (ADS)

Stefanutti, E.; Sierra, A.; Miocchi, P.; Massimi, L.; Brun, F.; Maugeri, L.; Bukreeva, I.; Nurmi, A.; Begani Provinciali, G.; Tromba, G.; Gröhn, O.; Giove, F.; Cedola, A.; Fratini, M.

2018-03-01

Synchrotron X-ray Phase Contrast micro-Tomography (SXrPCμT) is a powerful tool in the investigation of biological tissues, including the central nervous system (CNS), and it allows to simultaneously detect the vascular and neuronal network avoiding contrast agents or destructive sample preparations. However, specific sample preparation procedures aimed to optimize the achievable contrast- and signal-to-noise ratio (CNR and SNR, respectively) are required. Here we report and discuss the effects of perfusion with two different fixative agents (ethanol and paraformaldehyde) and with a widely used contrast medium (MICROFIL®) on mouse spinal cord. As a main result, we found that ethanol enhances contrast at the grey/white matter interface and increases the contrast in correspondence of vascular features and fibres, thus providing an adequate spatial resolution to visualise the vascular network at the microscale. On the other hand, ethanol is known to induce tissue dehydration, likely reducing cell dimensions below the spatial resolution limit imposed by the experimental technique. Nonetheless, neurons remain well visible using either perfused paraformaldehyde or MICROFIL® compound, as these latter media do not affect tissues with dehydration effects. Paraformaldehyde appears as the best compromise: it is not a contrast agent, like MICROFIL®, but it is less invasive than ethanol and permits to visualise well both cells and blood vessels. However, a quantitative estimation of the relative grey matter volume of each sample has led us to conclude that no significant alterations in the grey matter extension compared to the white matter occur as a consequence of the perfusion procedures tested in this study.

Synthesis of Programmable Main-chain Liquid-crystalline Elastomers Using a Two-stage Thiol-acrylate Reaction.

PubMed

Saed, Mohand O; Torbati, Amir H; Nair, Devatha P; Yakacki, Christopher M

2016-01-19

This study presents a novel two-stage thiol-acrylate Michael addition-photopolymerization (TAMAP) reaction to prepare main-chain liquid-crystalline elastomers (LCEs) with facile control over network structure and programming of an aligned monodomain. Tailored LCE networks were synthesized using routine mixing of commercially available starting materials and pouring monomer solutions into molds to cure. An initial polydomain LCE network is formed via a self-limiting thiol-acrylate Michael-addition reaction. Strain-to-failure and glass transition behavior were investigated as a function of crosslinking monomer, pentaerythritol tetrakis(3-mercaptopropionate) (PETMP). An example non-stoichiometric system of 15 mol% PETMP thiol groups and an excess of 15 mol% acrylate groups was used to demonstrate the robust nature of the material. The LCE formed an aligned and transparent monodomain when stretched, with a maximum failure strain over 600%. Stretched LCE samples were able to demonstrate both stress-driven thermal actuation when held under a constant bias stress or the shape-memory effect when stretched and unloaded. A permanently programmed monodomain was achieved via a second-stage photopolymerization reaction of the excess acrylate groups when the sample was in the stretched state. LCE samples were photo-cured and programmed at 100%, 200%, 300%, and 400% strain, with all samples demonstrating over 90% shape fixity when unloaded. The magnitude of total stress-free actuation increased from 35% to 115% with increased programming strain. Overall, the two-stage TAMAP methodology is presented as a powerful tool to prepare main-chain LCE systems and explore structure-property-performance relationships in these fascinating stimuli-sensitive materials.

78 FR 7858 - Proposed Collection; Comment Request for Information Collection tools.

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-04

... Foreign Corporation; Form 13614-NR, Volunteer Return Preparation Critical Intake Sheet-NR; and REG-209006... min. Estimated Total Annual Burden Hours: 35. (2) Title: Volunteer Return Preparation Critical Intake... nonresident alien VITA sites by volunteers to gather information--relevant to tax preparation--from taxpayer's...

Fluid sampling tool

DOEpatents

Garcia, Anthony R.; Johnston, Roger G.; Martinez, Ronald K.

2000-01-01

A fluid-sampling tool for obtaining a fluid sample from a container. When used in combination with a rotatable drill, the tool bores a hole into a container wall, withdraws a fluid sample from the container, and seals the borehole. The tool collects fluid sample without exposing the operator or the environment to the fluid or to wall shavings from the container.

Novel Sample-handling Approach for XRD Analysis with Minimal Sample Preparation

NASA Technical Reports Server (NTRS)

Sarrazin, P.; Chipera, S.; Bish, D.; Blake, D.; Feldman, S.; Vaniman, D.; Bryson, C.

2004-01-01

Sample preparation and sample handling are among the most critical operations associated with X-ray diffraction (XRD) analysis. These operations require attention in a laboratory environment, but they become a major constraint in the deployment of XRD instruments for robotic planetary exploration. We are developing a novel sample handling system that dramatically relaxes the constraints on sample preparation by allowing characterization of coarse-grained material that would normally be impossible to analyze with conventional powder-XRD techniques.

Proteomics Analysis of Tissue Samples Reveals Changes in Mitochondrial Protein Levels in Parathyroid Hyperplasia over Adenoma

PubMed Central

AKPINAR, GURLER; KASAP, MURAT; CANTURK, NUH ZAFER; ZULFIGAROVA, MEHIN; ISLEK, EYLÜL ECE; GULER, SERTAC ATA; SIMSEK, TURGAY; CANTURK, ZEYNEP

2017-01-01

Background/Aim: To unveil the pathophysiology of primary hyperparathyroidism, molecular details of parathyroid hyperplasia and adenoma have to be revealed. Such details will provide the tools necessary for differentiation of these two look-alike diseases. Therefore, in the present study, a comparative proteomic study using postoperative tissue samples from the parathyroid adenoma and parathyroid hyperplasia patients was performed. Materials and Methods: Protein extracts were prepared from tissue samples (n=8 per group). Protein pools were created for each group and subjected to DIGE and conventional 2DE. Following image analysis, spots representing the differentially regulated proteins were excised from the and used for identification via MALDI-TOF/TOF analysis. Results: The identities of 40 differentially-expressed proteins were revealed. Fourteen of these proteins were over-expressed in the hyperplasia while 26 of them were over-expressed in the adenoma. Conclusion: Most proteins found to be over-expressed in the hyperplasia samples were mitochondrial, underlying the importance of the mitochondrial activity as a potential biomarker for differentiation of parathyroid hyperplasia from adenoma. PMID:28446534

Use of XPS to clarify the Hall coefficient sign variation in thin niobium layers buried in silicon

NASA Astrophysics Data System (ADS)

Demchenko, Iraida N.; Lisowski, Wojciech; Syryanyy, Yevgen; Melikhov, Yevgen; Zaytseva, Iryna; Konstantynov, Pavlo; Chernyshova, Maryna; Cieplak, Marta Z.

2017-03-01

Si/Nb/Si trilayers formed with 9.5 and 1.3 nm thick niobium layer buried in amorphous silicon were prepared by magnetron sputtering and studied using XPS depth-profile techniques in order to investigate the change of Hall coefficient sign with thickness. The analysis of high-resolution (HR) XPS spectra revealed that the thicker layer sample has sharp top interface and metallic phase of niobium, thus holes dominate the transport. In contrast, the analysis indicates that the thinner layer sample has a Nb-rich mixed alloy formation at the top interface. The authors suggest that the main effect leading to a change of sign of the Hall coefficient for the thinner layer sample (which is negative contrary to the positive sign for the thicker layer sample) may be related to strong boundary scattering enhanced by the presence of silicon ions in the layer close to the interface/s. The depth-profile reconstruction was performed by SESSA software tool confirming that it can be reliably used for quantitative analysis/interpretation of experimental XPS data.

Detection of Salmonella enterica Serovar Typhimurium by Using a Rapid, Array-Based Immunosensor

PubMed Central

Taitt, Chris Rowe; Shubin, Yura S.; Angel, Roselina; Ligler, Frances S.

2004-01-01

The multianalyte array biosensor (MAAB) is a rapid analysis instrument capable of detecting multiple analytes simultaneously. Rapid (15-min), single-analyte sandwich immunoassays were developed for the detection of Salmonella enterica serovar Typhimurium, with a detection limit of 8 × 104 CFU/ml; the limit of detection was improved 10-fold by lengthening the assay protocol to 1 h. S. enterica serovar Typhimurium was also detected in the following spiked foodstuffs, with minimal sample preparation: sausage, cantaloupe, whole liquid egg, alfalfa sprouts, and chicken carcass rinse. Cross-reactivity tests were performed with Escherichia coli and Campylobacter jejuni. To determine whether the MAAB has potential as a screening tool for the diagnosis of asymptomatic Salmonella infection of poultry, chicken excretal samples from a private, noncommercial farm and from university poultry facilities were tested. While the private farm excreta gave rise to signals significantly above the buffer blanks, none of the university samples tested positive for S. enterica serovar Typhimurium without spiking; dose-response curves of spiked excretal samples from university-raised poultry gave limits of detection of 8 × 103 CFU/g. PMID:14711637

Measurement of stable isotopic enrichment and concentration of long-chain fatty acyl-carnitines in tissue by HPLC-MS.

PubMed

Sun, Dayong; Cree, Melanie G; Zhang, Xiao-Jun; Bøersheim, Elisabet; Wolfe, Robert R

2006-02-01

We have developed a new method for the simultaneous measurements of stable isotopic tracer enrichments and concentrations of individual long-chain fatty acyl-carnitines in muscle tissue using ion-pairing high-performance liquid chromatography-electrospray ionization quadrupole mass spectrometry in the selected ion monitoring (SIM) mode. Long-chain fatty acyl-carnitines were extracted from frozen muscle tissue samples by acetonitrile/methanol. Baseline separation was achieved by reverse-phase HPLC in the presence of the volatile ion-pairing reagent heptafluorobutyric acid. The SIM capability of a single quadrupole mass analyzer allows further separation of the ions of interest from the sample matrixes, providing very clean total and selected ion chromatograms that can be used to calculate the stable isotopic tracer enrichment and concentration of long-chain fatty acyl-carnitines in a single analysis. The combination of these two separation techniques greatly simplifies the sample preparation procedure and increases the detection sensitivity. Applying this protocol to biological muscle samples proves it to be a very sensitive, accurate, and precise analytical tool.

Accelerated bridge construction (ABC) decision making and economic modeling tool.

DOT National Transportation Integrated Search

2011-12-01

In this FHWA-sponsored pool funded study, a set of decision making tools, based on the Analytic Hierarchy Process (AHP) was developed. This tool set is prepared for transportation specialists and decision-makers to determine if ABC is more effective ...

Nontargeted, Rapid Screening of Extra Virgin Olive Oil Products for Authenticity Using Near-Infrared Spectroscopy in Combination with Conformity Index and Multivariate Statistical Analyses.

PubMed

Karunathilaka, Sanjeewa R; Kia, Ali-Reza Fardin; Srigley, Cynthia; Chung, Jin Kyu; Mossoba, Magdi M

2016-10-01

A rapid tool for evaluating authenticity was developed and applied to the screening of extra virgin olive oil (EVOO) retail products by using Fourier-transform near infrared (FT-NIR) spectroscopy in combination with univariate and multivariate data analysis methods. Using disposable glass tubes, spectra for 62 reference EVOO, 10 edible oil adulterants, 20 blends consisting of EVOO spiked with adulterants, 88 retail EVOO products and other test samples were rapidly measured in the transmission mode without any sample preparation. The univariate conformity index (CI) and the multivariate supervised soft independent modeling of class analogy (SIMCA) classification tool were used to analyze the various olive oil products which were tested for authenticity against a library of reference EVOO. Better discrimination between the authentic EVOO and some commercial EVOO products was observed with SIMCA than with CI analysis. Approximately 61% of all EVOO commercial products were flagged by SIMCA analysis, suggesting that further analysis be performed to identify quality issues and/or potential adulterants. Due to its simplicity and speed, FT-NIR spectroscopy in combination with multivariate data analysis can be used as a complementary tool to conventional official methods of analysis to rapidly flag EVOO products that may not belong to the class of authentic EVOO. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Degradation of metallic materials studied by correlative tomography

NASA Astrophysics Data System (ADS)

Burnett, T. L.; Holroyd, N. J. H.; Lewandowski, J. J.; Ogurreck, M.; Rau, C.; Kelley, R.; Pickering, E. J.; Daly, M.; Sherry, A. H.; Pawar, S.; Slater, T. J. A.; Withers, P. J.

2017-07-01

There are a huge array of characterization techniques available today and increasingly powerful computing resources allowing for the effective analysis and modelling of large datasets. However, each experimental and modelling tool only spans limited time and length scales. Correlative tomography can be thought of as the extension of correlative microscopy into three dimensions connecting different techniques, each providing different types of information, or covering different time or length scales. Here the focus is on the linking of time lapse X-ray computed tomography (CT) and serial section electron tomography using the focussed ion beam (FIB)-scanning electron microscope to study the degradation of metals. Correlative tomography can provide new levels of detail by delivering a multiscale 3D picture of key regions of interest. Specifically, the Xe+ Plasma FIB is used as an enabling tool for large-volume high-resolution serial sectioning of materials, and also as a tool for preparation of microscale test samples and samples for nanoscale X-ray CT imaging. The exemplars presented illustrate general aspects relating to correlative workflows, as well as to the time-lapse characterisation of metal microstructures during various failure mechanisms, including ductile fracture of steel and the corrosion of aluminium and magnesium alloys. Correlative tomography is already providing significant insights into materials behaviour, linking together information from different instruments across different scales. Multiscale and multifaceted work flows will become increasingly routine, providing a feed into multiscale materials models as well as illuminating other areas, particularly where hierarchical structures are of interest.

Racial Differences in Test Preparation Strategies: A Commentary on "Shadow Education, American Style: Test Preparation, the SAT and College Enrollment"

ERIC Educational Resources Information Center

Alon, Sigal

2010-01-01

Claudia Buchmann, Dennis Condron and Vincent Roscigno's study, titled "Shadow Education, American Style: Test Preparation, the SAT and College Enrollment," demonstrates that vigorous use of expensive test preparation tools, such as private classes and tutors, significantly boosts scores on standardized exams such as the SAT or ACT. This…

Sampling hazelnuts for aflatoxin: uncertainty associated with sampling, sample preparation, and analysis.

PubMed

Ozay, Guner; Seyhan, Ferda; Yilmaz, Aysun; Whitaker, Thomas B; Slate, Andrew B; Giesbrecht, Francis

2006-01-01

The variability associated with the aflatoxin test procedure used to estimate aflatoxin levels in bulk shipments of hazelnuts was investigated. Sixteen 10 kg samples of shelled hazelnuts were taken from each of 20 lots that were suspected of aflatoxin contamination. The total variance associated with testing shelled hazelnuts was estimated and partitioned into sampling, sample preparation, and analytical variance components. Each variance component increased as aflatoxin concentration (either B1 or total) increased. With the use of regression analysis, mathematical expressions were developed to model the relationship between aflatoxin concentration and the total, sampling, sample preparation, and analytical variances. The expressions for these relationships were used to estimate the variance for any sample size, subsample size, and number of analyses for a specific aflatoxin concentration. The sampling, sample preparation, and analytical variances associated with estimating aflatoxin in a hazelnut lot at a total aflatoxin level of 10 ng/g and using a 10 kg sample, a 50 g subsample, dry comminution with a Robot Coupe mill, and a high-performance liquid chromatographic analytical method are 174.40, 0.74, and 0.27, respectively. The sampling, sample preparation, and analytical steps of the aflatoxin test procedure accounted for 99.4, 0.4, and 0.2% of the total variability, respectively.

Near infrared photoluminescence properties of porous silicon prepared under the influence of light illumination

NASA Astrophysics Data System (ADS)

Hamadeh, H.; Naddaf, M.; Jazmati, A.

2008-12-01

Porous silicon (PS) has been prepared by anodic etching of boron doped silicon under the influence of monochromatic light illumination. The optical properties of the PS samples have been investigated using temperature dependent photoluminescence (PL) spectroscopy. An overall enhancement of the infrared luminescence yield is caused by the light illumination. In the visible spectral range, changes at the low energy side of the broad PL band were observed. In the near infrared spectral range, a new PL band at 850 nm, which is strongly correlated with light illumination, was detected. The new PL band disappears once blue light is used, whereas an increase in its intensity is observed, when the etching is performed under the illumination of light with wavelengths close to the band gap. By increasing the temperature, the 850 nm transition band grows at the expense of the main near infrared transition at 1100 nm. The recombination characteristics of this PL band are indicative of its extrinsic nature. The macroscopic morphology shows strong dependence on the wavelength of the illumination light. Photoassisted preparation could provide a tool for the control of the optical and structural properties of PS.

7 CFR 61.34 - Drawing and preparation of sample.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 3 2010-01-01 2010-01-01 false Drawing and preparation of sample. 61.34 Section 61.34 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Cottonseed Samplers § 61.34 Drawing and preparation of sample. Each licensed cottonseed sampler shall draw...

CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING AIR SAMPLES FOR ANALYSIS OF POLAR PERSISTENT ORGANIC POLLUTANTS (SOP-5.13)

EPA Science Inventory

The method for extracting and preparing indoor and outdoor air samples for analysis of polar persistent organic pollutants is summarized in this SOP. It covers the preparation of samples that are to be analyzed by gas chromatography/mass spectrometry.

40 CFR 761.392 - Preparing validation study samples.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Preparing validation study samples..., AND USE PROHIBITIONS Comparison Study for Validating a New Performance-Based Decontamination Solvent Under § 761.79(d)(4) § 761.392 Preparing validation study samples. (a)(1) To validate a procedure to...

40 CFR 761.392 - Preparing validation study samples.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Preparing validation study samples..., AND USE PROHIBITIONS Comparison Study for Validating a New Performance-Based Decontamination Solvent Under § 761.79(d)(4) § 761.392 Preparing validation study samples. (a)(1) To validate a procedure to...

Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria.

PubMed

Lucchi, Naomi W; Demas, Allison; Narayanan, Jothikumar; Sumari, Deborah; Kabanywanyi, Abdunoor; Kachur, S Patrick; Barnwell, John W; Udhayakumar, Venkatachalam

2010-10-29

Molecular diagnostic methods can complement existing tools to improve the diagnosis of malaria. However, they require good laboratory infrastructure thereby restricting their use to reference laboratories and research studies. Therefore, adopting molecular tools for routine use in malaria endemic countries will require simpler molecular platforms. The recently developed loop-mediated isothermal amplification (LAMP) method is relatively simple and can be improved for better use in endemic countries. In this study, we attempted to improve this method for malaria diagnosis by using a simple and portable device capable of performing both the amplification and detection (by fluorescence) of LAMP in one platform. We refer to this as the RealAmp method. Published genus-specific primers were used to test the utility of this method. DNA derived from different species of malaria parasites was used for the initial characterization. Clinical samples of P. falciparum were used to determine the sensitivity and specificity of this system compared to microscopy and a nested PCR method. Additionally, directly boiled parasite preparations were compared with a conventional DNA isolation method. The RealAmp method was found to be simple and allowed real-time detection of DNA amplification. The time to amplification varied but was generally less than 60 minutes. All human-infecting Plasmodium species were detected. The sensitivity and specificity of RealAmp in detecting P. falciparum was 96.7% and 91.7% respectively, compared to microscopy and 98.9% and 100% respectively, compared to a standard nested PCR method. In addition, this method consistently detected P. falciparum from directly boiled blood samples. This RealAmp method has great potential as a field usable molecular tool for diagnosis of malaria. This tool can provide an alternative to conventional PCR based diagnostic methods for field use in clinical and operational programs.

Program Quality in Leadership Preparation Programs: An Assessment Tool

ERIC Educational Resources Information Center

Campanotta, Linda; Simpson, Patricia; Newton, Jodi

2018-01-01

The leadership of school principals is widely considered to have a strong influence on the quality of teaching and learning in schools. University-based leadership preparation programs have the responsibility for preparing candidates to serve as school principals and in other leadership capacities. This qualitative study of educational leadership…

7 CFR 27.89 - Expenses; inspection; sampling.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Expenses; inspection; sampling. 27.89 Section 27.89... Micronaire § 27.89 Expenses; inspection; sampling. Expense of inspection and sampling, the preparation of the... Office, the expense of inspection, sampling, preparation of samples, and delivery of the samples to the...

7 CFR 27.89 - Expenses; inspection; sampling.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Expenses; inspection; sampling. 27.89 Section 27.89... Micronaire § 27.89 Expenses; inspection; sampling. Expense of inspection and sampling, the preparation of the... Office, the expense of inspection, sampling, preparation of samples, and delivery of the samples to the...

Coaxial cable stripper for confined areas

NASA Technical Reports Server (NTRS)

Brown, J. D.; Lipscomb, W. G.

1968-01-01

Manual coaxial cable stripper quickly and accurately prepares a coaxial cable in a confined area. With this tool, preparation time is greatly reduced, and a completely inexperienced technician can perform the operation.

The Factor Structure and Psychometric Properties of the Persian Version of the Revised Prenatal Coping Inventory (Nu-PCI).

PubMed

Faramarzi, Mahbobeh; Pasha, Hajar; Khafri, Sorayya; Heidary, Shima

2017-03-01

Familiarity with coping strategies is essential for stress management during pregnancy. The Revised Prenatal Coping Inventory (Nu-PCI) was developed to assess coping strategies during pregnancy. This study aimed to assess the factor structure and psychometric properties of the Persian version of the Nu-PCI. After forward-backward translation, the Nu-PCI was administered to 210 pregnant women who were enrolled in two teaching referral clinics in the North of the Islamic Republic of Iran (Babol). The participants completed the Persian Nu-PCI and Ways of Coping Questionnaire (WCQ), which was used to determine the validity of the Persian Nu-PCI. To test construct validity of the Persian Nu-PCI, a principal components factor analysis was performed. Principal components analysis with varimax rotation showed a best fitting 3-factor structure similar to the original with three coping subscales: planning-preparation, avoidance, and spiritual-positive coping. The Persian Nu-PCI was internally consistent and within the acceptable range (α=0.89-0.97). The alpha coefficients for the Nu-PCI and the subscales of planning-preparation, avoidance, and spiritual-positive coping were high. Test-retest coefficients for the Nu-PCI and subscales were 0.98-0.99. The Nu-PCI and its subscales correlated with the WCQ in the entire sample and within each trimester. The Persian version of the Nu-PCI and the subscales of planning-preparation, avoidance, and spiritual-positive coping represent the first reliable standardized tool for measuring coping strategies during pregnancy in the Islamic Republic of Iran. Therefore, it can be applied as a quick and accurate preliminary screening tool for evaluating coping strategies throughout pregnancy in clinics and other medical and research settings.

Outlining and dictating scientific manuscripts is a useful method for health researchers: A focus group interview.

PubMed

Andresen, Kristoffer; Laursen, Jannie; Rosenberg, Jacob

2018-01-01

Young researchers may experience difficulties when writing scientific articles for publication in biomedical journals. Various methods may facilitate the writing process including outlining the paper before the actual writing and using dictation instead of writing the first draft. The aim of this study was to investigate the experiences and difficulties for young, experienced researchers when writing articles using a detailed outline and dictation of the first draft. We used qualitative focus group interviews and the study was reported according to the COnsolidated criteria for REporting Qualitative research guideline. Participants were sampled from a group of researchers participating in a writing retreat/course. The interviews were recorded on a digital recorder and transcribed. The text was analyzed according to content analysis and coded and condensed into themes and subthemes. Groups of participants were added until data saturation was reached. A total of 14 researchers participated (9 women and 5 men). Their clinical experience was median (range) of 6 (1-11) years since graduation from medical school. Two themes arose during the analyses of the data: "Process guidance with the outline as the map" and "arrival at dictation." The outline was used in the preparation phase leading up to the day of dictation and was used in collaboration with co-authors and supervisors. The participants found it to be a useful tool for preparing the manuscript and dictating their initial first full draft. Experienced young researchers found beneficial effects of using a structured outline to prepare for dictation of scientific articles. The outline was a tool that would develop in close collaboration with co-authors and mentors. With dictation, a full first draft of a manuscript can be produced in a few hours. Participants positively evaluated this structured and reproducible way of producing scientific articles.

Fabrication and Characterization of High Temperature Resin/Carbon Nanofiber Composites

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Working, Dennis C.; Criss, Jim M.; Siochi, Emilie J.; Connell, John W.

2005-01-01

Multifunctional composites present a route to structural weight reduction. Nanoparticles such as carbon nanofibers (CNF) provide a compromise as a lower cost nanosize reinforcement that yields a desirable combination of properties. Blends of PETI-330 and CNFs were prepared and characterized to investigate the potential of CNF composites as a high performance structural medium. Dry mixing techniques were employed and the effect of CNF loading level on melt viscosity was determined. The resulting powders were characterized for degree of mixing, thermal and rheological properties. Based on the characterization results, samples containing 30 and 40 wt% CNF were scaled up to approx.300 g and used to fabricate moldings 10.2 cm x 15.2 cm x 0.32 cm thick. The moldings were fabricated by injecting the mixtures at 260-280 C into a stainless steel tool followed by curing for 1 h at 371 C. The tool was designed to impart high shear during the process in an attempt to achieve some alignment of CNFs in the flow direction. Moldings were obtained that were subsequently characterized for thermal, mechanical and electrical properties. The degree of dispersion and alignment of CNFs were investigated using high-resolution scanning electron microscopy. The preparation and preliminary characterization of PETI-330/CNF composites are discussed. Keywords: resins, carbon nanofibers, scanning electron microscopy, electrical properties, thermal conductivity,injection

Critical evaluation of reverse engineering tool Imagix 4D!

PubMed

Yadav, Rashmi; Patel, Ravindra; Kothari, Abhay

2016-01-01

The comprehension of legacy codes is difficult to understand. Various commercial reengineering tools are available that have unique working styles, and are equipped with their inherent capabilities and shortcomings. The focus of the available tools is in visualizing static behavior not the dynamic one. Therefore, it is difficult for people who work in software product maintenance, code understanding reengineering/reverse engineering. Consequently, the need for a comprehensive reengineering/reverse engineering tool arises. We found the usage of Imagix 4D to be good as it generates the maximum pictorial representations in the form of flow charts, flow graphs, class diagrams, metrics and, to a partial extent, dynamic visualizations. We evaluated Imagix 4D with the help of a case study involving a few samples of source code. The behavior of the tool was analyzed on multiple small codes and a large code gcc C parser. Large code evaluation was performed to uncover dead code, unstructured code, and the effect of not including required files at preprocessing level. The utility of Imagix 4D to prepare decision density and complexity metrics for a large code was found to be useful in getting to know how much reengineering is required. At the outset, Imagix 4D offered limitations in dynamic visualizations, flow chart separation (large code) and parsing loops. The outcome of evaluation will eventually help in upgrading Imagix 4D and posed a need of full featured tools in the area of software reengineering/reverse engineering. It will also help the research community, especially those who are interested in the realm of software reengineering tool building.

Imaging Gallium Nitride High Electron Mobility Transistors to Identify Point Defects

DTIC Science & Technology

2014-03-01

streamline the sample preparation procedure to maximize the yield of successful samples to be analyzed chemically in an energy dispersive spectrometry...transmission electron microscope (STEM), sample preparation 15. NUMBER OF PAGES 103 16. PRICE CODE 17. SECURITY CLASSIFICATION OF REPORT...Computer Engineering iii THIS PAGE INTENTIONALLY LEFT BLANK iv ABSTRACT The purpose of this thesis is to streamline the sample preparation

Using Isotope Ratio Infrared Spectrometer to determine δ13C and δ18O of carbonate samples

NASA Astrophysics Data System (ADS)

Smajgl, Danijela; Stöbener, Nils; Mandic, Magda

2017-04-01

The isotopic composition of calcifying organisms is a key tool for reconstruction past seawater temperature and water chemistry. Therefore stable carbon and oxygen isotopes (δ13C and δ18O) in carbonates have been widely used for reconstruction of paleoenvironments. Precise and accurate determination of isotopic composition of carbon (13C) and oxygen (18O) from carbonate sample with proper referencing and data evaluation algorithm presents a challenge for scientists. Mass spectrometry was the only widely used technique for this kind of analysis, but recent advances make laser based spectroscopy a viable alternative. The Thermo Scientific Delta Ray Isotope Ratio Infrared Spectrometer (IRIS) analyzer with the Universal Reference Interface (URI) Connect is one of those alternatives and with TELEDYNE Cetac ASX-7100 autosampler extends the traditional offerings with a system of high precision and throughput of samples. To establish precision and accuracy of measurements and also to develop optimal sample preparation method for measurements with Delta Ray IRIS and URI Connect, IAEA reference materials were used. Preparation is similar to a Gas Bench II method. Carbonate material is added into the vials, flushed with CO2 free synthetic air and acidified with few droplets of 104% H3PO4. Sample amount used for analysis can be as low as 200 μg. Samples are measured after acidification and equilibration time of one hour at 70°C. The CO2 gas generated by reaction is flushed into the variable volume inside the URI Connect through the Nafion based built-in water trap. For this step, carrier gas (CO2 free air) is used to flush the gas from the vial into the variable volume with a maximum volume of 100 ml. A small amount of the sample is then used for automatic concentration determination present in the variable volume. The Thermo Scientific Qtegra Software automatically adjusts any additional dilution of the sample to achieve the desired concentration (usually 400 ppm) in the analyzer. As part of the workflow, reference gas measurements are regularly measured at the same concentration as the sample to allow for automatic drift and linearity correction. With described sample preparation and measurement method, samples are measured with standard deviation less than 0.1‰ δ13C and δ18O, respectively and accuracy of <0.01‰. The system can measure up to 100 samples per day. Equivalent of about 80 µg of pure CO2 gas is needed to complete an analysis. Due to it's small weight and robustness, sample analysis can be performed in the field. Applying new technology of Isotope Ratio Infrared Spectrometers in environmental and paleoenvironmental research can extend the knowledge of complex seawater history and CO2 cycle.

Utility of Cytospin and Cell block Technology in Evaluation of Body Fluids and Urine Samples: A Comparative Study

PubMed Central

Qamar, Irmeen; Rehman, Suhailur; Mehdi, Ghazala; Maheshwari, Veena; Ansari, Hena A.; Chauhan, Sunanda

2018-01-01

Background: Cytologic examination of body fluids commonly involves the use of direct or sediment smears, cytocentrifuge preparations, membrane filter preparations, or cell block sections. Cytospin and cell block techniques are extremely useful in improving cell yield of thin serous effusions and urine samples, and ensure high diagnostic efficacy. Materials and Methods: We studied cytospin preparations and cell block sections prepared from 180 samples of body fluids and urine samples to compare the relative efficiency of cell retrieval, preservation of cell morphology, ease of application of special stains, and diagnostic efficacy. Samples were collected and processed to prepare cytospin smears and cell block sections. Results: We observed that overall, cell yield and preservation of individual cell morphology were better in cytospin preparations as compared to cell blocks, while preservation of architectural pattern was better in cell block sections. The number of suspicious cases also decreased on cell block sections, with increased detection of malignancy. It was difficult to prepare cell blocks from urine samples due to low cellularity. Conclusions: Cytospin technology is a quick, efficient, and cost-effective method of increasing cell yield in hypocellular samples, with better preservation of cell morphology. Cell blocks are better prepared from high cellularity fluids; however, tissue architecture is better studied, with improved rate of diagnosis and decrease in ambiguous results. Numerous sections can be prepared from a small amount of material. Special stains and immunochemical stains can be easily applied to cell blocks. It also provides a source of archival material. PMID:29643653

Coring Sample Acquisition Tool

NASA Technical Reports Server (NTRS)

Haddad, Nicolas E.; Murray, Saben D.; Walkemeyer, Phillip E.; Badescu, Mircea; Sherrit, Stewart; Bao, Xiaoqi; Kriechbaum, Kristopher L.; Richardson, Megan; Klein, Kerry J.

2012-01-01

A sample acquisition tool (SAT) has been developed that can be used autonomously to sample drill and capture rock cores. The tool is designed to accommodate core transfer using a sample tube to the IMSAH (integrated Mars sample acquisition and handling) SHEC (sample handling, encapsulation, and containerization) without ever touching the pristine core sample in the transfer process.

High-throughput automated microfluidic sample preparation for accurate microbial genomics

PubMed Central

Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B.; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P.; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C.

2017-01-01

Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications. PMID:28128213

Behavior of Aluminum in Solid Propellant Combustion

DTIC Science & Technology

1982-06-01

dry pressing 30% Valley Met H- 30 aluminum, 7% carnauba wax , and 63% 100 P AP. One sample was prepared using as received H-30, a second sample used pre...34propellant" formulations. The formulations included dry pressed AP/AI, and AP/AI/ Wax samples. Sandwiches were also prepared consisting of an aluminum...Binder flame instead of by aluminum exposure during accumulate break-up. Combustion of AP/AI/ Wax Samples A set of propellant samples were prepared by

Experimental Evaluation of the Tools of the Mind Pre-K Curriculum. Fidelity of Implementation Technical Report. Working Paper

ERIC Educational Resources Information Center

Meador, Deanna; Nesbitt, Kimberly; Farran, Dale

2015-01-01

The "Experimental Evaluation of the Tools of the Mind Pre-K Curriculum" study was designed to compare the effectiveness of the "Tools of the Mind" ("Tools") curriculum to the curricula the school system is currently using in enhancing children's self-regulation skills and their academic preparation for kindergarten.…

A comparison of sample preparation strategies for biological tissues and subsequent trace element analysis using LA-ICP-MS.

PubMed

Bonta, Maximilian; Török, Szilvia; Hegedus, Balazs; Döme, Balazs; Limbeck, Andreas

2017-03-01

Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) is one of the most commonly applied methods for lateral trace element distribution analysis in medical studies. Many improvements of the technique regarding quantification and achievable lateral resolution have been achieved in the last years. Nevertheless, sample preparation is also of major importance and the optimal sample preparation strategy still has not been defined. While conventional histology knows a number of sample pre-treatment strategies, little is known about the effect of these approaches on the lateral distributions of elements and/or their quantities in tissues. The technique of formalin fixation and paraffin embedding (FFPE) has emerged as the gold standard in tissue preparation. However, the potential use for elemental distribution studies is questionable due to a large number of sample preparation steps. In this work, LA-ICP-MS was used to examine the applicability of the FFPE sample preparation approach for elemental distribution studies. Qualitative elemental distributions as well as quantitative concentrations in cryo-cut tissues as well as FFPE samples were compared. Results showed that some metals (especially Na and K) are severely affected by the FFPE process, whereas others (e.g., Mn, Ni) are less influenced. Based on these results, a general recommendation can be given: FFPE samples are completely unsuitable for the analysis of alkaline metals. When analyzing transition metals, FFPE samples can give comparable results to snap-frozen tissues. Graphical abstract Sample preparation strategies for biological tissues are compared with regard to the elemental distributions and average trace element concentrations.

Dental Students' Perceptions of Digital Assessment Software for Preclinical Tooth Preparation Exercises.

PubMed

Park, Carly F; Sheinbaum, Justin M; Tamada, Yasushi; Chandiramani, Raina; Lian, Lisa; Lee, Cliff; Da Silva, John; Ishikawa-Nagai, Shigemi

2017-05-01

Objective self-assessment is essential to learning and continued competence in dentistry. A computer-assisted design/computer-assisted manufacturing (CAD/CAM) learning software (prepCheck, Sirona) allows students to objectively assess their performance in preclinical prosthodontics. The aim of this study was to evaluate students' perceptions of CAD/CAM learning software for preclinical prosthodontics exercises. In 2014, all third-year dental students at Harvard School of Dental Medicine (n=36) were individually instructed by a trained faculty member in using prepCheck. Each student completed a preclinical formative exercise (#18) and summative examination (#30) for ceramometal crown preparation and evaluated the preparation using five assessment tools (reduction, margin width, surface finish, taper, and undercut) in prepCheck. The students then rated each of the five tools for usefulness, user-friendliness, and frequency of use on a scale from 1=lowest to 5=highest. Faculty members graded the tooth preparations as pass (P), marginal-pass (MP), or fail (F). The survey response rate was 100%. The tools for undercut and taper had the highest scores for usefulness, user-friendliness, and frequency of use. The reduction tool score was significantly lower in all categories (p<0.01). There were significant differences in usefulness (p<0.05) and user-friendliness (p<0.05) scores among the P, MP, and F groups. These results suggest that the prepCheck taper and undercut tools were useful for the students' learning process in a preclinical exercise. The students' perceptions of prepCheck and their preclinical performance were related, and those students who performed poorest rated the software as significantly more useful.

A comprehensive investigation of mechanical and acoustic modifications in wood treated with high doses of gamma radiation for sterilization

NASA Astrophysics Data System (ADS)

Ricci, G.; Regazzi, R.; Tositti, L.; Barbaresi, L.; Liverani, A.; Mostacci, D.

2016-09-01

Gamma radiation is a most efficient tool for sterilization, and especially effective on infesting pest, as it also kills the eggs. However, the large radiation doses entailed (tens of kilogray and up) might have adverse effects on desirable properties of wood. Of particular interest are the acoustic properties, as radiation would otherwise appear to be the perfect tool for the sterilization of musical instruments. The present investigation was aimed at determining the effects on acoustic and mechanical properties. Samples of four different wood species - poplar, oak, fir and maple - were prepared selecting wood completely free of visually detectable defects and subjected to incremental dose levels, reaching 25, 50, 100 and 200 kGy. A further set of samples were subjected to 200 kGy in one go, to investigate possible differences with incremental doses. Irradiation was conducted at an industrial irradiation facility, operating with a cobalt-60 source of the order of the megacurie. The mechanical tests were designed to measure the elastic modulus and the bending strength as well as force and displacement at rupture. Acoustic tests were developed specifically for this experiment, with the purpose of measuring the vibrational modes of the samples. All the samples exhibited discoloration and noticeable changes in properties: rigidity increased, load to rupture decreased, the material became more brittle and effects increased with increasing dose. The acoustic tests revealed a shift toward higher frequencies of all the vibrational modes, and this was exhibited for all wood species investigated. No meaningful difference was observed between the samples treated with incremental doses up to a total of 200 kGy and with a single 200 kGy dose.

Proteomic analysis and comparison of the biopsy and autopsy specimen of human brain temporal lobe.

PubMed

He, Sizhi; Wang, Qingsong; He, Jintang; Pu, Hai; Yang, Wei; Ji, Jianguo

2006-09-01

The proteomic study on human temporal lobe can help us to understand the physiological function of CNS in normal as well as in pathological state. Proteomic tools are potent for the assessment of protein stability post mortem. In this pilot study, the human temporal lobe biopsy specimen with chronic pharmacoresistant temporal lobe epilepsy (TLE) and autopsy specimen in control were separated by 2-DE. Using MALDI-TOF-MS and MS/MS, 375 protein spots were identified which were the products of 267 genes. Six down-regulated and 23 up-regulated protein spots in the autopsy specimen were ascertained after the gel image analysis with the ImageMaster software. A number of proteins that include neurotransmitter metabolic and glycolytic enzymes, cytoprotective proteins and cytoskeleton were found decreased while the precursor of apolipoprotein A-I increased in the TLE brain. We tried several methods to prepare the protein samples and found that DNase and RNase treatment, ultracentrifugation and Amersham clean-up kit purification can improve gel separation quality. This work optimized the sample preparation method and constructed a primary protein database of human temporal lobe and found some proteins with remarkable level change probably involved in the post-mortem process and chronic pharmacoresistant TLE pathogenesis.

Assessing the Potential of Metal-Assisted Imaging Mass Spectrometry in Cancer Research.

PubMed

Dufresne, M; Patterson, N H; Lauzon, N; Chaurand, P

2017-01-01

In the last decade, imaging mass spectrometry (IMS) has been the primary tool for biomolecular imaging. While it is possible to map a wide range of biomolecules using matrix-assisted laser desorption/ionization IMS ranging from high-molecular-weight proteins to small metabolites, more often than not only the most abundant easily ionisable species are detected. To better understand complex diseases such as cancer more specific and sensitive methods need to be developed to enable the detection of lower abundance molecules but also molecules that have yet to be imaged by IMS. In recent years, a big shift has occurred in the imaging community from developing wide reaching methods to developing targeted ones which increases sensitivity through the use of more specific sample preparations. This has been primarily marked by the advent of solvent-free matrix deposition methods for polar lipids, chemical derivatization for hormones and metabolites, and the use of alternative ionization agents for neutral lipids. In this chapter, we discuss two of the latest sample preparations which exploit the use of alternative ionization agents to enable the detection of certain classes of neutral lipids along with free fatty acids by high-sensitivity IMS as demonstrated within our lab. © 2017 Elsevier Inc. All rights reserved.

Evaluating the Aging of Multiple Emulsions Using Resonance-Enhanced Multiphoton Ionization Time-of-Flight Mass Spectrometry.

PubMed

Tsuda, Yukihiro; Uchimura, Tomohiro

2016-01-01

Resonance-enhanced multiphoton ionization time-of-flight mass spectrometry was applied to measurements of multiple emulsions with no pretreatment; a method for the quantitative evaluation of aging was proposed. We prepared water-in-oil-in-water (W/O/W) multiple emulsions containing toluene and m-phenylenediamine. The samples were measured immediately following both preparation and after having been stirred for 24 h. Time profiles of the peak areas for each analyte species were obtained, and several intense spikes for toluene could be detected from each sample after stirring, which suggests that the concentration of toluene in the middle phase had increased during stirring. On the other hand, in the case of a W/O/W multiple emulsion containing phenol and m-phenylenediamine, spikes for m-phenylenediamine, rather than phenol, were detected after stirring. In the present study, the time-profile data were converted into a scatter plot in order to quantitatively evaluate the aging. As a result, the ratio of the plots where strong signal intensities of toluene were detected increased from 8.4% before stirring to 33.2% after stirring for 24 h. The present method could be a powerful tool for evaluating multiple emulsions, such as studies on the kinetics of the encapsulation and release of active ingredients.

Optimization of programmed-temperature vaporization injection preparative capillary GC for compound specific radiocarbon analysis.

PubMed

Zhang, Xinyu; Zhao, Liang; Wang, Yexin; Xu, Yunping; Zhou, Liping

2013-07-01

Preparative capillary GC (PCGC) is a powerful tool for the separation and purification of compounds from any complex matrix, which can be used for compound-specific radiocarbon analysis. However, the effect of PCGC parameters on the trapping efficiency is not well understood. Here, we present a comprehensive study on the optimization of parameters based on 11 reference compounds with different physicochemical properties. Under the optimum conditions, the trapping efficiencies of these 11 compounds (including high-boiling-point n-hentriacontane and methyl lignocerate) are about 80% (60-89%). The isolation of target compounds from standard solutions, plant and soil samples demonstrates that our optimized method is applicable for different classes of compounds including n-alkanes, fatty acid esters, long-chain fatty alcohol esters, polycyclic aromatic hydrocarbons (PAHs) and steranes. By injecting 25 μL in large volume injection mode, over 100 μg, high purity (>90%) target compounds are harvested within 24 h. The recovery ranges of two real samples are about 70% (59.9-83.8%) and about 83% (77.2-88.5%), respectively. Compared to previous studies, our study makes significant improvement in the recovery of PCGC, which is important for its wide application in biogeochemistry, environmental sciences, and archaeology. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A C8-Modified Graphene@mSiO2 Composites Based Method for Quantification of Gallic Acid in Rat Plasma after Oral Administration of Changtai Granule and Its Application to Pharmacokinetics.

PubMed

Xu, Chen; Yu, Yingjia; Ling, Li; Wang, Yang; Zhang, Jundong; Li, Yan; Duan, Gengli

2017-01-01

A rapid, effective extraction technique has been established for measuring the gallic acid in rat plasma by using sandwich-structured graphene/mesoporous silica composites with C 8 -modified interior pore-walls as adsorbent. The unique characteristics of the graphene-silica composites excluded large molecules, like proteins, from the mesopore channels as a result of size exclusion effect, leading to a direct extraction of drug molecules from protein-rich biological samples such as plasma without any other pretreatment procedure. Followed by elution and centrifugation, the gallic acid-absorbed composites were rapidly isolated before LC-MS/MS. Serving as a reliable tool for analysis of Traditional Chinese Medicine: Changtai Granule, the newly developed method was fully validated and successfully applied in the pharmacokinetic study of gallic acid in rat plasma. Extraction recovery, matrix effect and stability were satisfactory in rat plasma. According to the results of pharmacokinetic studies, Changtai Granule exhibited greater adsorption, distribution and clearance properties of gallic acid in the treatment of ulcerative colitis. Hence, this study may offer a valuable alternative to simplify and speed up sample preparation, and be useful for clinical studies of related preparations.

Single hair analysis of small molecules using MALDI-triple quadrupole MS imaging and LC-MS/MS: investigations on opportunities and pitfalls.

PubMed

Poetzsch, Michael; Steuer, Andrea E; Roemmelt, Andreas T; Baumgartner, Markus R; Kraemer, Thomas

2014-12-02

Single hair analysis normally requires extensive sample preparation microscale protocols including time-consuming steps like segmentation and extraction. Matrix assisted laser desorption and ionization mass spectrometric imaging (MALDI-MSI) was shown to be an alternative tool in single hair analysis, but still, questions remain. Therefore, an investigation of MALDI-MSI in single hair analysis concerning the extraction process, usage of internal standard (IS), and influences on the ionization processes were systematically investigated to enable the reliable application to hair analysis. Furthermore, single dose detection, quantitative correlation to a single hair, and hair strand LC-MS/MS results were performed, and the performance was compared to LC-MS/MS single hair monitoring. The MALDI process was shown to be independent from natural hair color and not influenced by the presence of melanin. Ionization was shown to be reproducible along and in between different hair samples. MALDI image intensities in single hair and hair snippets showed good semiquantitative correlation to zolpidem hair concentrations obtained from validated routine LC-MS/MS methods. MALDI-MSI is superior to LC-MS/MS analysis when a fast, easy, and cheap sample preparation is necessary, whereas LC-MS/MS showed higher sensitivity with the ability of single dose detection for zolpidem. MALDI-MSI and LC-MS/MS segmental single hair analysis showed good correlation, and both are suitable for consumption monitoring of drugs of abuse with a high time resolution.

Advances in Small Particle Handling of Astromaterials in Preparation for OSIRIS-REx and Hayabusa2: Initial Developments

NASA Technical Reports Server (NTRS)

Snead, C. J.; McCubbin, F. M.; Nakamura-Messenger, K.; Righter, K.

2018-01-01

The Astromaterials Acquisition and Curation office at NASA Johnson Space Center has established an Advanced Curation program that is tasked with developing procedures, technologies, and data sets necessary for the curation of future astromaterials collections as envisioned by NASA exploration goals. One particular objective of the Advanced Curation program is the development of new methods for the collection, storage, handling and characterization of small (less than 100 micrometer) particles. Astromaterials Curation currently maintains four small particle collections: Cosmic Dust that has been collected in Earth's stratosphere by ER2 and WB-57 aircraft, Comet 81P/Wild 2 dust returned by NASA's Stardust spacecraft, interstellar dust that was returned by Stardust, and asteroid Itokawa particles that were returned by the JAXA's Hayabusa spacecraft. NASA Curation is currently preparing for the anticipated return of two new astromaterials collections - asteroid Ryugu regolith to be collected by Hayabusa2 spacecraft in 2021 (samples will be provided by JAXA as part of an international agreement), and asteroid Bennu regolith to be collected by the OSIRIS-REx spacecraft and returned in 2023. A substantial portion of these returned samples are expected to consist of small particle components, and mission requirements necessitate the development of new processing tools and methods in order to maximize the scientific yield from these valuable acquisitions. Here we describe initial progress towards the development of applicable sample handling methods for the successful curation of future small particle collections.

Validation of Methods to Assess the Immunoglobulin Gene Repertoire in Tissues Obtained from Mice on the International Space Station.

PubMed

Rettig, Trisha A; Ward, Claire; Pecaut, Michael J; Chapes, Stephen K

2017-07-01

Spaceflight is known to affect immune cell populations. In particular, splenic B cell numbers decrease during spaceflight and in ground-based physiological models. Although antibody isotype changes have been assessed during and after space flight, an extensive characterization of the impact of spaceflight on antibody composition has not been conducted in mice. Next Generation Sequencing and bioinformatic tools are now available to assess antibody repertoires. We can now identify immunoglobulin gene- segment usage, junctional regions, and modifications that contribute to specificity and diversity. Due to limitations on the International Space Station, alternate sample collection and storage methods must be employed. Our group compared Illumina MiSeq sequencing data from multiple sample preparation methods in normal C57Bl/6J mice to validate that sample preparation and storage would not bias the outcome of antibody repertoire characterization. In this report, we also compared sequencing techniques and a bioinformatic workflow on the data output when we assessed the IgH and Igκ variable gene usage. This included assessments of our bioinformatic workflow on Illumina HiSeq and MiSeq datasets and is specifically designed to reduce bias, capture the most information from Ig sequences, and produce a data set that provides other data mining options. We validated our workflow by comparing our normal mouse MiSeq data to existing murine antibody repertoire studies validating it for future antibody repertoire studies.

Usability of Immunohistochemistry in Forensic Samples With Varying Decomposition.

PubMed

Lesnikova, Iana; Schreckenbach, Marc Niclas; Kristensen, Maria Pihlmann; Papanikolaou, Liv Lindegaard; Hamilton-Dutoit, Stephen

2018-05-24

Immunohistochemistry (IHC) is an important diagnostic tool in anatomic and surgical pathology but is used less frequently in forensic pathology. Degradation of tissue because of postmortem decomposition is believed to be a major limiting factor, although it is unclear what impact such degradation actually has on IHC staining validity. This study included 120 forensic autopsy samples of liver, lung, and brain tissues obtained for diagnostic purposes. The time from death to autopsy ranged between 1 and more than 14 days. Samples were prepared using the tissue microarray technique. The antibodies chosen for the study included KL1 (for staining bile duct epithelium), S100 (for staining glial cells and myelin), vimentin (for endothelial cells in cerebral blood vessels), and CD45 (for pulmonary lymphocytes). Slides were evaluated by light microscopy. Immunohistochemistry reactions were scored according to a system based on the extent and intensity of the positive stain. An overall correlation between the postmortem interval and the IHC score for all tissue samples was found. Samples from decedents with a postmortem interval of 1 to 3 days showed positive staining with all antibodies, whereas samples from decedents with a longer postmortem interval showed decreased staining rates. Our results suggest that IHC analysis can be successfully used for postmortem diagnosis in a range of autopsy samples showing lesser degrees of decomposition.

Preparation of Chemical Samples On Relevant Surfaces Using Inkjet Technology

DTIC Science & Technology

2013-04-01

PREPARATION OF CHEMICAL SAMPLES ON RELEVANT SURFACES USING INKJET TECHNOLOGY...2012 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Preparation of Chemical Samples on Relevant Surfaces Using Inkjet Technology 5b. GRANT NUMBER...SUBJECT TERMS Surface detection Inkjet Simulant deposition 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF

Airborne Observation of the Hayabusa Sample Return Capsule Re-Entry

NASA Technical Reports Server (NTRS)

Grinstead, Jay H.; Jenniskens, Peter; Cassell, Alan M.; Albers, James; Winter, Michael W.

2011-01-01

NASA Ames Research Center and the SETI Institute collaborated on an effort to observe the Earth re-entry of the Japan Aerospace Exploration Agency's Hayabusa sample return capsule. Hayabusa was an asteroid exploration mission that retrieved a sample from the near-Earth asteroid Itokawa. Its sample return capsule re-entered over the Woomera Prohibited Area in southern Australia on June 13, 2010. Being only the third sample return mission following NASA's Genesis and Stardust missions, Hayabusa's return was a rare opportunity to collect aerothermal data from an atmospheric entry capsule returning at superorbital speeds. NASA deployed its DC-8 airborne laboratory and a team of international researchers to Australia for the re-entry. For approximately 70 seconds, spectroscopic and radiometric imaging instruments acquired images and spectra of the capsule, its wake, and destructive re-entry of the spacecraft bus. Once calibrated, spectra of the capsule will be interpreted to yield data for comparison with and validation of high fidelity and engineering simulation tools used for design and development of future atmospheric entry system technologies. A brief summary of the Hayabusa mission, the preflight preparations and observation mission planning, mission execution, and preliminary spectral data are documented.

Spectroscopy applied to feed additives of the European Union Reference Laboratory: a valuable tool for traceability.

PubMed

Omar, Jone; Slowikowski, Boleslaw; Boix, Ana; von Holst, Christoph

2017-08-01

Feed additives need to be authorised to be placed on the market according to Regulation (EU) No. 1831/2003. Next to laying down the procedural requirements, the regulation creates the European Union Reference Laboratory for Feed Additives (EURL-FA) and requires that applicants send samples to the EURL-FA. Once authorised, the characteristics of the marketed feed additives should correspond to those deposited in the sample bank of the EURL-FA. For this purpose, the submitted samples were subjected to near-infrared (NIR) and Raman spectroscopy for spectral characterisation. These techniques have the valuable potential of characterising the feed additives in a non-destructive manner without any complicated sample preparation. This paper describes the capability of spectroscopy for a rapid characterisation of products to establish whether specific authorisation criteria are met. This study is based on the analysis of feed additive samples from different categories and functional groups, namely products containing (1) selenium, (2) zinc and manganese, (3) vitamins and (4) essential oils such as oregano and thyme oil. The use of chemometrics turned out to be crucial, especially in cases where the differentiation of spectra by visual inspection was very difficult.

Rapid, all-optical crystal orientation imaging of two-dimensional transition metal dichalcogenide monolayers

DOE Office of Scientific and Technical Information (OSTI.GOV)

David, Sabrina N.; Zhai, Yao; van der Zande, Arend M.

Two-dimensional (2D) atomic materials such as graphene and transition metal dichalcogenides (TMDCs) have attracted significant research and industrial interest for their electronic, optical, mechanical, and thermal properties. While large-area crystal growth techniques such as chemical vapor deposition have been demonstrated, the presence of grain boundaries and orientation of grains arising in such growths substantially affect the physical properties of the materials. There is currently no scalable characterization method for determining these boundaries and orientations over a large sample area. We here present a second-harmonic generation based microscopy technique for rapidly mapping grain orientations and boundaries of 2D TMDCs. We experimentallymore » demonstrate the capability to map large samples to an angular resolution of ±1° with minimal sample preparation and without involved analysis. A direct comparison of the all-optical grain orientation maps against results obtained by diffraction-filtered dark-field transmission electron microscopy plus selected-area electron diffraction on identical TMDC samples is provided. This rapid and accurate tool should enable large-area characterization of TMDC samples for expedited studies of grain boundary effects and the efficient characterization of industrial-scale production techniques.« less

Digital gene expression analysis with sample multiplexing and PCR duplicate detection: A straightforward protocol.

PubMed

Rozenberg, Andrey; Leese, Florian; Weiss, Linda C; Tollrian, Ralph

2016-01-01

Tag-Seq is a high-throughput approach used for discovering SNPs and characterizing gene expression. In comparison to RNA-Seq, Tag-Seq eases data processing and allows detection of rare mRNA species using only one tag per transcript molecule. However, reduced library complexity raises the issue of PCR duplicates, which distort gene expression levels. Here we present a novel Tag-Seq protocol that uses the least biased methods for RNA library preparation combined with a novel approach for joint PCR template and sample labeling. In our protocol, input RNA is fragmented by hydrolysis, and poly(A)-bearing RNAs are selected and directly ligated to mixed DNA-RNA P5 adapters. The P5 adapters contain i5 barcodes composed of sample-specific (moderately) degenerate base regions (mDBRs), which later allow detection of PCR duplicates. The P7 adapter is attached via reverse transcription with individual i7 barcodes added during the amplification step. The resulting libraries can be sequenced on an Illumina sequencer. After sample demultiplexing and PCR duplicate removal with a free software tool we designed, the data are ready for downstream analysis. Our protocol was tested on RNA samples from predator-induced and control Daphnia microcrustaceans.

Humidity-controlled preparation of frozen-hydrated biological samples for cryogenic coherent x-ray diffraction microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takayama, Yuki; Nakasako, Masayoshi; RIKEN Harima Institute/SPring-8, 1-1-1 Kouto, Mikaduki, Sayo, Hyogo 679-5148

2012-05-15

Coherent x-ray diffraction microscopy (CXDM) has the potential to visualize the structures of micro- to sub-micrometer-sized biological particles, such as cells and organelles, at high resolution. Toward advancing structural studies on the functional states of such particles, here, we developed a system for the preparation of frozen-hydrated biological samples for cryogenic CXDM experiments. The system, which comprised a moist air generator, microscope, micro-injector mounted on a micromanipulator, custom-made sample preparation chamber, and flash-cooling device, allowed for the manipulation of sample particles in the relative humidity range of 20%-94%rh at 293 K to maintain their hydrated and functional states. Here, wemore » report the details of the system and the operation procedure, including its application to the preparation of a frozen-hydrated chloroplast sample. Sample quality was evaluated through a cryogenic CXDM experiment conducted at BL29XUL of SPring-8. Taking the performance of the system and the quality of the sample, the system was suitable to prepare frozen-hydrated biological samples for cryogenic CXDM experiments.« less

Advanced Curation of Current and Future Extraterrestrial Samples

NASA Technical Reports Server (NTRS)

Allen, Carlton C.

2013-01-01

Curation of extraterrestrial samples is the critical interface between sample return missions and the international research community. Curation includes documentation, preservation, preparation, and distribution of samples. The current collections of extraterrestrial samples include: Lunar rocks / soils collected by the Apollo astronauts Meteorites, including samples of asteroids, the Moon, and Mars "Cosmic dust" (asteroid and comet particles) collected by high-altitude aircraft Solar wind atoms collected by the Genesis spacecraft Comet particles collected by the Stardust spacecraft Interstellar dust collected by the Stardust spacecraft Asteroid particles collected by the Hayabusa spacecraft These samples were formed in environments strikingly different from that on Earth. Terrestrial contamination can destroy much of the scientific significance of many extraterrestrial materials. In order to preserve the research value of these precious samples, contamination must be minimized, understood, and documented. In addition the samples must be preserved - as far as possible - from physical and chemical alteration. In 2011 NASA selected the OSIRIS-REx mission, designed to return samples from the primitive asteroid 1999 RQ36 (Bennu). JAXA will sample C-class asteroid 1999 JU3 with the Hayabusa-2 mission. ESA is considering the near-Earth asteroid sample return mission Marco Polo-R. The Decadal Survey listed the first lander in a Mars sample return campaign as its highest priority flagship-class mission, with sample return from the South Pole-Aitken basin and the surface of a comet among additional top priorities. The latest NASA budget proposal includes a mission to capture a 5-10 m asteroid and return it to the vicinity of the Moon as a target for future sampling. Samples, tools, containers, and contamination witness materials from any of these missions carry unique requirements for acquisition and curation. Some of these requirements represent significant advances over methods currently used. New analytical and screening techniques will increase the value of current sample collections. Improved web-based tools will make information on all samples more accessible to researchers and the public. Advanced curation of current and future extraterrestrial samples includes: Contamination Control - inorganic / organic Temperature of preservation - subfreezing / cryogenic Non-destructive preliminary examination - X-ray tomography / XRF mapping / Raman mapping Microscopic samples - handling / sectioning / transport Special samples - unopened lunar cores Informatics - online catalogs / community-based characterization.

Preparation of bone samples in the Gliwice Radiocarbon Laboratory for AMS radiocarbon dating.

PubMed

Piotrowska, N; Goslar, T

2002-12-01

In the Gliwice Radiocarbon Laboratory, a system for preparation of samples for AMS dating has been built. At first it was used to produce graphite targets from plant macrofossils and sediments. In this study we extended its capabilities with the preparation of bones. We dealt with 3 methods; the first was the classical Longin method of collagen extraction, the second one included additional treatment of powdered bone in alkali solution, while in the third one carboxyl carbon was separated from amino acids obtained after hydrolysis of protein. The suitability of the methods was tested on 2 bone samples. Most of our samples gave ages > 40 kyr BP, suggesting good performance of the adapted methods, except for one sample prepared with simple Longin method. For routine preparation of bones we chose the Longin method with additional alkali treatment.

Finding Relevant Parameters for the Thin-film Photovoltaic Cells Production Process with the Application of Data Mining Methods.

PubMed

Ulaczyk, Jan; Morawiec, Krzysztof; Zabierowski, Paweł; Drobiazg, Tomasz; Barreau, Nicolas

2017-09-01

A data mining approach is proposed as a useful tool for the control parameters analysis of the 3-stage CIGSe photovoltaic cell production process, in order to find variables that are the most relevant for cell electric parameters and efficiency. The analysed data set consists of stage duration times, heater power values as well as temperatures for the element sources and the substrate - there are 14 variables per sample in total. The most relevant variables of the process have been found based on the so-called random forest analysis with the application of the Boruta algorithm. 118 CIGSe samples, prepared at Institut des Matériaux Jean Rouxel, were analysed. The results are close to experimental knowledge on the CIGSe cells production process. They bring new evidence to production parameters of new cells and further research. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chemical and biological threat-agent detection using electrophoresis-based lab-on-a-chip devices.

PubMed

Borowsky, Joseph; Collins, Greg E

2007-10-01

The ability to separate complex mixtures of analytes has made capillary electrophoresis (CE) a powerful analytical tool since its modern configuration was first introduced over 25 years ago. The technique found new utility with its application to the microfluidics based lab-on-a-chip platform (i.e., microchip), which resulted in ever smaller footprints, sample volumes, and analysis times. These features, coupled with the technique's potential for portability, have prompted recent interest in the development of novel analyzers for chemical and biological threat agents. This article will comment on three main areas of microchip CE as applied to the separation and detection of threat agents: detection techniques and their corresponding limits of detection, sampling protocol and preparation time, and system portability. These three areas typify the broad utility of lab-on-a-chip for meeting critical, present-day security, in addition to illustrating areas wherein advances are necessary.

A user's guide to localization-based super-resolution fluorescence imaging.

PubMed

Dempsey, Graham T

2013-01-01

Advances in far-field fluorescence microscopy over the past decade have led to the development of super-resolution imaging techniques that provide more than an order of magnitude improvement in spatial resolution compared to conventional light microscopy. One such approach, called Stochastic Optical Reconstruction Microscopy (STORM) uses the sequential, nanometer-scale localization of individual fluorophores to reconstruct a high-resolution image of a structure of interest. This is an attractive method for biological investigation at the nanoscale due to its relative simplicity, both conceptually and practically in the laboratory. Like most research tools, however, the devil is in the details. The aim of this chapter is to serve as a guide for applying STORM to the study of biological samples. This chapter will discuss considerations for choosing a photoswitchable fluorescent probe, preparing a sample, selecting hardware for data acquisition, and collecting and analyzing data for image reconstruction. Copyright © 2013 Elsevier Inc. All rights reserved.

[Applications of MALDI-TOF-MS in clinical microbiology laboratory].

PubMed

Carbonnelle, Etienne; Nassif, Xavier

2011-10-01

For twenty years, mass spectrometry (MS) has emerged as a particularly powerful tool for analysis and characterization of proteins in research. It is only recently that this technology, especially MALDI-TOF-MS (Matrix Assisted Laser Desorption Ionization Time-Of-Flight) has entered the field of routine microbiology. This method has proven to be reliable and safe for the identification of bacteria, yeasts, filamentous fungi and dermatophytes. MALDI-TOF-MS is a rapid, precise and cost-effective method for identification, compared to conventional phenotypic techniques or molecular biology. Its ability to analyse whole microorganisms with few sample preparation has greatly reduced the time to identification (1-2 min). Furthermore, this technology can be used to identify bacteria directly from clinical samples as blood culture bottles or urines. Future applications will be developed in order to provide direct information concerning virulence or resistance protein markers. © 2011 médecine/sciences – Inserm / SRMS.

From pixel to voxel: a deeper view of biological tissue by 3D mass spectral imaging

PubMed Central

Ye, Hui; Greer, Tyler; Li, Lingjun

2011-01-01

Three dimensional mass spectral imaging (3D MSI) is an exciting field that grants the ability to study a broad mass range of molecular species ranging from small molecules to large proteins by creating lateral and vertical distribution maps of select compounds. Although the general premise behind 3D MSI is simple, factors such as choice of ionization method, sample handling, software considerations and many others must be taken into account for the successful design of a 3D MSI experiment. This review provides a brief overview of ionization methods, sample preparation, software types and technological advancements driving 3D MSI research of a wide range of low- to high-mass analytes. Future perspectives in this field are also provided to conclude that the positive and promises ever-growing applications in the biomedical field with continuous developments of this powerful analytical tool. PMID:21320052

Terahertz imaging applied to cancer diagnosis.

PubMed

Brun, M-A; Formanek, F; Yasuda, A; Sekine, M; Ando, N; Eishii, Y

2010-08-21

We report on terahertz (THz) time-domain spectroscopy imaging of 10 microm thick histological sections. The sections are prepared according to standard pathological procedures and deposited on a quartz window for measurements in reflection geometry. Simultaneous acquisition of visible images enables registration of THz images and thus the use of digital pathology tools to investigate the links between the underlying cellular structure and specific THz information. An analytic model taking into account the polarization of the THz beam, its incidence angle, the beam shift between the reference and sample pulses as well as multiple reflections within the sample is employed to determine the frequency-dependent complex refractive index. Spectral images are produced through segmentation of the extracted refractive index data using clustering methods. Comparisons of visible and THz images demonstrate spectral differences not only between tumor and healthy tissues but also within tumors. Further visualization using principal component analysis suggests different mechanisms as to the origin of image contrast.

The Importance of Experimental Design, Quality Assurance, and Control in Plant Metabolomics Experiments.

PubMed

Martins, Marina C M; Caldana, Camila; Wolf, Lucia Daniela; de Abreu, Luis Guilherme Furlan

2018-01-01

The output of metabolomics relies to a great extent upon the methods and instrumentation to identify, quantify, and access spatial information on as many metabolites as possible. However, the most modern machines and sophisticated tools for data analysis cannot compensate for inappropriate harvesting and/or sample preparation procedures that modify metabolic composition and can lead to erroneous interpretation of results. In addition, plant metabolism has a remarkable degree of complexity, and the number of identified compounds easily surpasses the number of samples in metabolomics analyses, increasing false discovery risk. These aspects pose a large challenge when carrying out plant metabolomics experiments. In this chapter, we address the importance of a proper experimental design taking into consideration preventable complications and unavoidable factors to achieve success in metabolomics analysis. We also focus on quality control and standardized procedures during the metabolomics workflow.

Automated payload and instruments for astrobiology research developed and studied by German medium-sized space industry in cooperation with European academia

NASA Astrophysics Data System (ADS)

Schulte, Wolfgang; Hofer, Stefan; Hofmann, Peter; Thiele, Hans; von Heise-Rotenburg, Ralf; Toporski, Jan; Rettberg, Petra

2007-06-01

For more than a decade Kayser-Threde, a medium-sized enterprise of the German space industry, has been involved in astrobiology research in partnership with a variety of scientific institutes from all over Europe. Previous projects include exobiology research platforms in low Earth orbit on retrievable carriers and onboard the Space Station. More recently, exobiology payloads for in situ experimentation on Mars have been studied by Kayser-Threde under ESA contracts, specifically the ExoMars Pasteur Payload. These studies included work on a sample preparation and distribution systems for Martian rock/regolith samples, instrument concepts such as Raman spectroscopy and a Life Marker Chip, advanced microscope systems as well as robotic tools for astrobiology missions. The status of the funded technical studies and major results are presented. The reported industrial work was funded by ESA and the German Aerospace Center (DLR).

Tools for phospho- and glycoproteomics of plasma membranes.

PubMed

Wiśniewski, Jacek R

2011-07-01

Analysis of plasma membrane proteins and their posttranslational modifications is considered as important for identification of disease markers and targets for drug treatment. Due to their insolubility in water, studying of plasma membrane proteins using mass spectrometry has been difficult for a long time. Recent technological developments in sample preparation together with important improvements in mass spectrometric analysis have facilitated analysis of these proteins and their posttranslational modifications. Now, large scale proteomic analyses allow identification of thousands of membrane proteins from minute amounts of sample. Optimized protocols for affinity enrichment of phosphorylated and glycosylated peptides have set new dimensions in the depth of characterization of these posttranslational modifications of plasma membrane proteins. Here, I summarize recent advances in proteomic technology for the characterization of the cell surface proteins and their modifications. In the focus are approaches allowing large scale mapping rather than analytical methods suitable for studying individual proteins or non-complex mixtures.

Rapid bacterial diagnostics via surface enhanced Raman microscopy.

PubMed

Premasiri, W R; Sauer-Budge, A F; Lee, J C; Klapperich, C M; Ziegler, L D

2012-06-01

There is a continuing need to develop new techniques for the rapid and specific identification of bacterial pathogens in human body fluids especially given the increasing prevalence of drug resistant strains. Efforts to develop a surface enhanced Raman spectroscopy (SERS) based approach, which encompasses sample preparation, SERS substrates, portable Raman microscopy instrumentation and novel identification software, are described. The progress made in each of these areas in our laboratory is summarized and illustrated by a spiked infectious sample for urinary tract infection (UTI) diagnostics. SERS bacterial spectra exhibit both enhanced sensitivity and specificity allowing the development of an easy to use, portable, optical platform for pathogen detection and identification. SERS of bacterial cells is shown to offer not only reproducible molecular spectroscopic signatures for analytical applications in clinical diagnostics, but also is a new tool for studying biochemical activity in real time at the outer layers of these organisms.

Effect Of Neodymium Substitution In Structural Characteristics Of Magnesium Ferrite

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thankachan, Smitha; Binu, P. J.; Xavier, Sheena

2011-10-20

The effect of Nd{sup 3+} substitution on the structural properties of Magnesium ferrite was studied in the series MgNd{sub x}Fe{sub 2-x}O{sub 4}, where x = 0 to 0.3 in steps of 0.05. The series was prepared by sol-gel technique which is one of the novel technique to prepare nanosized samples. Structural characterization was done using X-ray diffractometer and Fourier Transform Infrared Spectrometer. XRD analysis reveals the prepared samples are single phasic till x = 0.2. From x0 = .25, a secondary phase of iron neodymium oxide appears along with the spinel phase. Particle size calculation shows the prepared samples aremore » in the 9nm to 11 nm regime. Lattice parameter was found to increase with concentration of Nd. XRD and FTIR analysis confirmed spinel structure of the prepared samples. XRF result shows the expected composition of prepared samples. The frequency dependence of the dielectric constant in the range 100 Hz--120MHz was also studied« less

A 96-well screen filter plate for high-throughput biological sample preparation and LC-MS/MS analysis.

PubMed

Peng, Sean X; Cousineau, Martin; Juzwin, Stephen J; Ritchie, David M

2006-01-01

A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates.

prepare_taxa_charts.py: A Python program to automate generation of publication ready taxonomic pie chart images from QIIME.

PubMed

Lakhujani, Vijay; Badapanda, Chandan

2017-06-01

QIIME (Quantitative Insights Into Microbial Ecology) is one of the most popular open-source bioinformatics suite for performing metagenome, 16S rRNA amplicon and Internal Transcribed Spacer (ITS) data analysis. Although, it is very comprehensive and powerful tool, it lacks a method to provide publication ready taxonomic pie charts. The script plot_taxa_summary . py bundled with QIIME generate a html file and a folder containing taxonomic pie chart and legend as separate images. The images have randomly generated alphanumeric names. Therefore, it is difficult to associate the pie chart with the legend and the corresponding sample identifier. Even if the option to have the legend within the html file is selected while executing plot_taxa_summary . py , it is very tedious to crop a complete image (having both the pie chart and the legend) due to unequal image sizes. It requires a lot of time to manually prepare the pie charts for multiple samples for publication purpose. Moreover, there are chances of error while identifying the pie chart and legend pair due to random alphanumeric names of the images. To bypass all these bottlenecks and make this process efficient, we have developed a python based program, prepare_taxa_charts . py , to automate the renaming, cropping and merging of taxonomic pie chart and corresponding legend image into a single, good quality publication ready image. This program not only augments the functionality of plot_taxa_summary . py but is also very fast in terms of CPU time and user friendly.

High-quality cell block preparation from scraping of conventional cytology slide: a technical report on a modified cytoscrape cell block technique.

PubMed

Choi, Y I; Jakhongir, M; Choi, S J; Kim, L; Park, I S; Han, J Y; Kim, J M; Chu, Y C

2016-12-01

Immunocytochemistry (ICC) on formalin-fixed paraffin embedded cell blocks is an ancillary tool commonly recruited for differential diagnoses of fine needle aspiration cytology (FNAC) samples. However, the quality of conventional cell blocks in terms of adequate cellularity and evenness of distribution of cytologic material is not always satisfactory for ICC. We introduce a modified agarose-based cytoscrape cell block (CCB) technique that can be effectively used for the preparation of cell blocks from scrapings of conventional FNAC slides. A decoverslipped FNAC slide was mounted with a small amount of water. The cytological material was scraped off the slide into a tissue mold by scraping with a cell scraper. The cytoscrape material was pelleted by centrifugation and pre-embedded in ultra-low gelling temperature agarose and then re-embedded in conventional agarose. The final agarose gel disk was processed and embedded in paraffin. The quality of the ICC on the CCB sections was identical to that of the immunohistochemical stains on histological sections. By scrapping and harvesting the entirety of the cytological material off the cytology slide into a compact agarose cell button, we could avoid the risk of losing diagnostic material during the CCB preparation. This modified CCB technique enables concentration and focusing of minute material while maintaining the entire amount of the cytoscrape material on the viewing spot of the CCB sections. We believe this technique can be effectively used to improve the level of confidence in diagnosis of FNAC especially when the FNAC slides are the only sample available.

Tube Alinement for Machining

NASA Technical Reports Server (NTRS)

Garcia, J.

1984-01-01

Tool with stepped shoulders alines tubes for machining in preparation for welding. Alinement with machine tool axis accurate to within 5 mils (0.13mm) and completed much faster than visual setup by machinist.

Recent advances of mesoporous materials in sample preparation.

PubMed

Zhao, Liang; Qin, Hongqiang; Wu, Ren'an; Zou, Hanfa

2012-03-09

Sample preparation has been playing an important role in the analysis of complex samples. Mesoporous materials as the promising adsorbents have gained increasing research interest in sample preparation due to their desirable characteristics of high surface area, large pore volume, tunable mesoporous channels with well defined pore-size distribution, controllable wall composition, as well as modifiable surface properties. The aim of this paper is to review the recent advances of mesoporous materials in sample preparation with emphases on extraction of metal ions, adsorption of organic compounds, size selective enrichment of peptides/proteins, specific capture of post-translational peptides/proteins and enzymatic reactor for protein digestion. Copyright © 2011 Elsevier B.V. All rights reserved.

Sulfometuron methyl: Its use in forestry and potential phytotoxicity

Treesearch

Nathan D. Robertson; Anthony S. Davis

2010-01-01

Planting site preparation is a common practice used to enhance seedling establishment success. Site preparations include herbicide, fire, and mechanical methods. Studies designed to explore the use of herbicides as site preparation and release tools are common, and herbicides have shown their use in forestry to be logistically, economically, and ecologically...

40 CFR 761.323 - Sample preparation.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 761.323 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL... Remediation Waste Samples § 761.323 Sample preparation. (a) The comparison study requires analysis of a minimum of 10 samples weighing at least 300 grams each. Samples of PCB remediation waste used in the...

40 CFR 761.323 - Sample preparation.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 761.323 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL... Remediation Waste Samples § 761.323 Sample preparation. (a) The comparison study requires analysis of a minimum of 10 samples weighing at least 300 grams each. Samples of PCB remediation waste used in the...

40 CFR 761.323 - Sample preparation.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 761.323 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL... Remediation Waste Samples § 761.323 Sample preparation. (a) The comparison study requires analysis of a minimum of 10 samples weighing at least 300 grams each. Samples of PCB remediation waste used in the...

14C sample preparation for AMS microdosing studies at Lund University using online combustion and septa-sealed vials

NASA Astrophysics Data System (ADS)

Sydoff, Marie; Stenström, Kristina

2010-04-01

The Department of Physics at Lund University is participating in a European Union project called EUMAPP (European Union Microdose AMS Partnership Programme), in which sample preparation and accelerator mass spectrometry (AMS) measurements of biological samples from microdosing studies have been made. This paper describes a simplified method of converting biological samples to solid graphite for 14C analysis with AMS. The method is based on online combustion of the samples, and reduction of CO 2 in septa-sealed vials. The septa-sealed vials and disposable materials are used to eliminate sample cross-contamination. Measurements of ANU and Ox I standards show deviations of 2% and 3%, respectively, relative to reference values. This level of accuracy is sufficient for biological samples from microdosing studies. Since the method has very few handling steps from sample to graphite, the risk of failure during the sample preparation process is minimized, making the method easy to use in routine preparation of samples.

46 CFR 164.009-15 - Test procedure.

Code of Federal Regulations, 2010 CFR

2010-10-01

... material, is less than 47 mm, the specimens prepared consist of layers of the sample. (3) If the sample is a composite material and has a height that is not 50 ±3mm, the layers of the specimen prepared are proportional in thickness to the layers of the sample. (4) The top and bottom faces of each specimen prepared...

9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

Application of Multimedia Design Principles to Visuals Used in Course-Books: An Evaluation Tool

ERIC Educational Resources Information Center

Kuzu, Abdullah; Akbulut, Yavuz; Sahin, Mehmet Can

2007-01-01

This paper introduces an evaluation tool prepared to examine the quality of visuals in course-books. The tool is based on Mayer's Cognitive Theory of Multimedia Learning (i.e. Generative Theory) and its principles regarding the correct use of illustrations within text. The reason to generate the tool, the development process along with the…

Appendix W. Cost Analysis in Teacher Education Programs.

ERIC Educational Resources Information Center

Sell, G. Roger; And Others

This paper is an introduction to the basic cost-related tools available to management for planning, evaluating, and organizing resources for the purpose of achieving objectives within a teacher education preparation program. Three tools are presented in separate sections. Part I on the cost accounting tool for identifying, categorizing, and…

Experimental Evaluation of the Tools of the Mind Preschool Curriculum

ERIC Educational Resources Information Center

Wilson, Sandra Jo; Farran, Dale C.

2012-01-01

The aim of the "Tools of the Mind" prekindergarten curriculum is to enhance children's executive function skills within an instructional context that promotes the basic academic and social skills that prepare them for kindergarten and beyond. To investigate the effectiveness of "Tools" in achieving this aim, the authors are…

Integrating Computational Science Tools into a Thermodynamics Course

ERIC Educational Resources Information Center

Vieira, Camilo; Magana, Alejandra J.; García, R. Edwin; Jana, Aniruddha; Krafcik, Matthew

2018-01-01

Computational tools and methods have permeated multiple science and engineering disciplines, because they enable scientists and engineers to process large amounts of data, represent abstract phenomena, and to model and simulate complex concepts. In order to prepare future engineers with the ability to use computational tools in the context of…

Price and cost estimation

NASA Technical Reports Server (NTRS)

Stewart, R. D.

1979-01-01

Price and Cost Estimating Program (PACE II) was developed to prepare man-hour and material cost estimates. Versatile and flexible tool significantly reduces computation time and errors and reduces typing and reproduction time involved in preparation of cost estimates.

Pre-Mission Input Requirements to Enable Successful Sample Collection by A Remote Field/EVA Team

NASA Technical Reports Server (NTRS)

Cohen, B. A.; Lim, D. S. S.; Young, K. E.; Brunner, A.; Elphic, R. E.; Horne, A.; Kerrigan, M. C.; Osinski, G. R.; Skok, J. R.; Squyres, S. W.;

Analytical challenges for conducting rapid metabolism characterization for QIVIVE.

PubMed

Tolonen, Ari; Pelkonen, Olavi

2015-06-05

For quantitative in vitro-in vivo extrapolation (QIVIVE) of metabolism for the purposes of toxicokinetics prediction, a precise and robust analytical technique for identifying and measuring a chemical and its metabolites is an absolute prerequisite. Currently, high-resolution mass spectrometry (HR-MS) is a tool of choice for a majority of organic relatively lipophilic molecules, linked with a LC separation tool and simultaneous UV-detection. However, additional techniques such as gas chromatography, radiometric measurements and NMR, are required to cover the whole spectrum of chemical structures. To accumulate enough reliable and robust data for the validation of QIVIVE, there are some partially opposing needs: Detailed delineation of the in vitro test system to produce a reliable toxicokinetic measure for a studied chemical, and a throughput capacity of the in vitro set-up and the analytical tool as high as possible. We discuss current analytical challenges for the identification and quantification of chemicals and their metabolites, both stable and reactive, focusing especially on LC-MS techniques, but simultaneously attempting to pinpoint factors associated with sample preparation, testing conditions and strengths and weaknesses of a particular technique available for a particular task. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Monitoring of antisolvent crystallization of sodium scutellarein by combined FBRM-PVM-NIR.

PubMed

Liu, Xuesong; Sun, Di; Wang, Feng; Wu, Yongjiang; Chen, Yong; Wang, Longhu

2011-06-01

Antisolvent crystallization can be used as an alternative to cooling or evaporation for the separation and purification of solid product in the pharmaceutical industry. To improve the process understanding of antisolvent crystallization, the use of in-line tools is vital. In this study, the process analytical technology (PAT) tools including focused beam reflectance measurement (FBRM), particle video microscope (PVM), and near-infrared spectroscopy (NIRS) were utilized to monitor antisolvent crystallization of sodium scutellarein. FBRM was used to monitor chord count and chord length distribution of sodium scutellarein particles in the crystallizer, and PVM, as an in-line video camera, provided pictures imaging particle shape and dimension. In addition, a quantitative model of PLS was established by in-line NIRS to detect the concentration of sodium scutellarein in the solvent and good calibration statistics were obtained (r(2) = 0.976) with the residual predictive deviation value of 11.3. The discussion over sensitivities, strengths, and weaknesses of the PAT tools may be helpful in selection of suitable PAT techniques. These in-line techniques eliminate the need for sample preparation and offer a time-saving approach to understand and monitor antisolvent crystallization process. Copyright © 2011 Wiley-Liss, Inc.

Hekate: Software Suite for the Mass Spectrometric Analysis and Three-Dimensional Visualization of Cross-Linked Protein Samples

PubMed Central

2013-01-01

Chemical cross-linking of proteins combined with mass spectrometry provides an attractive and novel method for the analysis of native protein structures and protein complexes. Analysis of the data however is complex. Only a small number of cross-linked peptides are produced during sample preparation and must be identified against a background of more abundant native peptides. To facilitate the search and identification of cross-linked peptides, we have developed a novel software suite, named Hekate. Hekate is a suite of tools that address the challenges involved in analyzing protein cross-linking experiments when combined with mass spectrometry. The software is an integrated pipeline for the automation of the data analysis workflow and provides a novel scoring system based on principles of linear peptide analysis. In addition, it provides a tool for the visualization of identified cross-links using three-dimensional models, which is particularly useful when combining chemical cross-linking with other structural techniques. Hekate was validated by the comparative analysis of cytochrome c (bovine heart) against previously reported data.1 Further validation was carried out on known structural elements of DNA polymerase III, the catalytic α-subunit of the Escherichia coli DNA replisome along with new insight into the previously uncharacterized C-terminal domain of the protein. PMID:24010795

Coupling of metal-organic frameworks-containing monolithic capillary-based selective enrichment with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry for efficient analysis of protein phosphorylation.

PubMed

Li, Daojin; Yin, Danyang; Chen, Yang; Liu, Zhen

2017-05-19

Protein phosphorylation is a major post-translational modification, which plays a vital role in cellular signaling of numerous biological processes. Mass spectrometry (MS) has been an essential tool for the analysis of protein phosphorylation, for which it is a key step to selectively enrich phosphopeptides from complex biological samples. In this study, metal-organic frameworks (MOFs)-based monolithic capillary has been successfully prepared as an effective sorbent for the selective enrichment of phosphopeptides and has been off-line coupled with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for efficient analysis of phosphopeptides. Using š-casein as a representative phosphoprotein, efficient phosphorylation analysis by this off-line platform was verified. Phosphorylation analysis of a nonfat milk sample was also demonstrated. Through introducing large surface areas and highly ordered pores of MOFs into monolithic column, the MOFs-based monolithic capillary exhibited several significant advantages, such as excellent selectivity toward phosphopeptides, superb tolerance to interference and simple operation procedure. Because of these highly desirable properties, the MOFs-based monolithic capillary could be a useful tool for protein phosphorylation analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of heterogeneous gallstones using laser-induced breakdown spectroscopy (LIBS) and wavelength dispersive X-ray fluorescence (WD-XRF).

PubMed

Jaswal, Brij Bir S; Kumar, Vinay; Sharma, Jitendra; Rai, Pradeep K; Gondal, Mohammed A; Gondal, Bilal; Singh, Vivek K

2016-04-01

Laser-induced breakdown spectroscopy (LIBS) is an emerging analytical technique with numerous advantages such as rapidity, multi-elemental analysis, no specific sample preparation requirements, non-destructiveness, and versatility. It has been proven to be a robust elemental analysis tool attracting interest because of being applied to a wide range of materials including biomaterials. In this paper, we have performed spectroscopic studies on gallstones which are heterogeneous in nature using LIBS and wavelength dispersive X-ray fluorescence (WD-XRF) techniques. It has been observed that the presence and relative concentrations of trace elements in different kind of gallstones (cholesterol and pigment gallstones) can easily be determined using LIBS technique. From the experiments carried out on gallstones for trace elemental mapping and detection, it was found that LIBS is a robust tool for such biomedical applications. The stone samples studied in the present paper were classified using the Fourier transform infrared (FTIR) spectroscopy. WD-XRF spectroscopy has been applied for the qualitative and quantitative analysis of major and trace elements present in the gallstone which was compared with the LIBS data. The results obtained in the present paper show interesting prospects for LIBS and WD-XRF to study cholelithiasis better.

La 2-xSr xCuO 4-δ superconducting samples prepared by the wet-chemical method

NASA Astrophysics Data System (ADS)

Loose, A.; Gonzalez, J. L.; Lopez, A.; Borges, H. A.; Baggio-Saitovitch, E.

2009-10-01

In this work, we report on the physical properties of good-quality polycrystalline superconducting samples of La 2-xSr xCu 1-yZn yO 4-δ ( y=0, 0.02) prepared by a wet-chemical method, focusing on the temperature dependence of the critical current. Using the wet-chemical method, we were able to produce samples with improved homogeneity compared to the solid-state method. A complete set of samples with several carrier concentrations, ranging from the underdoped (strontium concentration x≈0.05) to the highly overdoped ( x≈0.25) region, were prepared and investigated. The X-ray diffraction analysis, zero-field cooling magnetization and electrical resistivity measurements were reported on earlier. The structural parameters of the prepared samples seem to be slightly modified by the preparation method and their critical temperatures were lower than reported in the literature. The temperature dependence of the critical current was explained by a theoretical model which took the granular structure of the samples into account.

The minimum information required for a glycomics experiment (MIRAGE) project: sample preparation guidelines for reliable reporting of glycomics datasets.

PubMed

Struwe, Weston B; Agravat, Sanjay; Aoki-Kinoshita, Kiyoko F; Campbell, Matthew P; Costello, Catherine E; Dell, Anne; Ten Feizi; Haslam, Stuart M; Karlsson, Niclas G; Khoo, Kay-Hooi; Kolarich, Daniel; Liu, Yan; McBride, Ryan; Novotny, Milos V; Packer, Nicolle H; Paulson, James C; Rapp, Erdmann; Ranzinger, Rene; Rudd, Pauline M; Smith, David F; Tiemeyer, Michael; Wells, Lance; York, William S; Zaia, Joseph; Kettner, Carsten

2016-09-01

The minimum information required for a glycomics experiment (MIRAGE) project was established in 2011 to provide guidelines to aid in data reporting from all types of experiments in glycomics research including mass spectrometry (MS), liquid chromatography, glycan arrays, data handling and sample preparation. MIRAGE is a concerted effort of the wider glycomics community that considers the adaptation of reporting guidelines as an important step towards critical evaluation and dissemination of datasets as well as broadening of experimental techniques worldwide. The MIRAGE Commission published reporting guidelines for MS data and here we outline guidelines for sample preparation. The sample preparation guidelines include all aspects of sample generation, purification and modification from biological and/or synthetic carbohydrate material. The application of MIRAGE sample preparation guidelines will lead to improved recording of experimental protocols and reporting of understandable and reproducible glycomics datasets. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Investigation with respect to content and general properties of physics 10 textbook in accordance with the 2013 secondary school physics curriculum

NASA Astrophysics Data System (ADS)

Kavcar, Nevzat; Korkmaz, Cihan

2017-02-01

Purpose of this work is to determine the physics teacher candidates' views on Physics 10 textbook' content and general properties suitable to the 2013 Secondary School Physics Curriculum. 23 teacher candidates at 2014-2015 school year constituted the sampling of the study in which scanning model based on qualitative research technique was used by performing document analysis. Data collection tool of the research was the files prepared with 51 and nine open ended questions including the subject content and general properties of the textbook. It was concluded that the textbook was sufficient for being life context -based, language, activity-based and student-centered approximation, development of social and inquiry skills, and was insufficient for referring educational gains of the Curriculum, involving activities, projects and homework about application. Activities and applications about affective area, such tools for assessment and evaluation practices as concept map, concept network and semantic analysis table may be involved in the textbook.

X-Ray Crystallography as a Tool to Determine Three-Dimensional Structures of Commercial Enzymes Subjected to Treatment in Pressurized Fluids.

PubMed

Feiten, Mirian Cristina; Di Luccio, Marco; Santos, Karine F; de Oliveira, Débora; Oliveira, J Vladimir

2017-06-01

The study of enzyme function often involves a multi-disciplinary approach. Several techniques are documented in the literature towards determining secondary and tertiary structures of enzymes, and X-ray crystallography is the most explored technique for obtaining three-dimensional structures of proteins. Knowledge of three-dimensional structures is essential to understand reaction mechanisms at the atomic level. Additionally, structures can be used to modulate or improve functional activity of enzymes by the production of small molecules that act as substrates/cofactors or by engineering selected mutants with enhanced biological activity. This paper presentes a short overview on how to streamline sample preparation for crystallographic studies of treated enzymes. We additionally revise recent developments on the effects of pressurized fluid treatment on activity and stability of commercial enzymes. Future directions and perspectives on the the role of crystallography as a tool to access the molecular mechanisms underlying enzymatic activity modulation upon treatment in pressurized fluids are also addressed.

A Clomazone Immunoassay to Study the Environmental Fate of the Herbicide in Rice (Oryza sativa) Agriculture

PubMed Central

Carlomagno, M.; Mathó, C.; Cantou, G.; Sanborn, J. R.; Last, J. A.; Hammock, B. D.; Roel, A.; González, D.; González-Sapienza, G.

2010-01-01

The environmental impact of rice agriculture is poorly studied in developing countries, mainly, due to limitations of the analytical capacity. Here we report the development of a clomazone ELISA as a fast and cost-effective tool to monitor the dissipation of this herbicide along the harvest. Antibodies were prepared using different strategies of hapten conjugation, and the best hapten/antibody pair was selected. It proved to be a reliable tool to measure the herbicide in the 2.0-20 ng/mL range in field samples, with excellent correlation with HPLC results. The assay was used to study the dissipation of the herbicide in floodwater of experimental rice paddies in Uruguay. Large differences in the residual amount of herbicide were observed depending on the flooding practices. Due to its robustness and simplicity, the assay may be useful to delineate and monitor management practices that can contribute to minimizing the release of the herbicide in the environment. PMID:20302341

Advanced 3D Characterization and Reconstruction of Reactor Materials FY16 Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fromm, Bradley; Hauch, Benjamin; Sridharan, Kumar

2016-12-01

A coordinated effort to link advanced materials characterization methods and computational modeling approaches is critical to future success for understanding and predicting the behavior of reactor materials that operate at extreme conditions. The difficulty and expense of working with nuclear materials have inhibited the use of modern characterization techniques on this class of materials. Likewise, mesoscale simulation efforts have been impeded due to insufficient experimental data necessary for initialization and validation of the computer models. The objective of this research is to develop methods to integrate advanced materials characterization techniques developed for reactor materials with state-of-the-art mesoscale modeling and simulationmore » tools. Research to develop broad-ion beam sample preparation, high-resolution electron backscatter diffraction, and digital microstructure reconstruction techniques; and methods for integration of these techniques into mesoscale modeling tools are detailed. Results for both irradiated and un-irradiated reactor materials are presented for FY14 - FY16 and final remarks are provided.« less

Non-Gated Laser Induced Breakdown Spectroscopy Provides a Powerful Segmentation Tool on Concomitant Treatment of Characteristic and Continuum Emission

PubMed Central

Dasari, Ramachandra Rao; Barman, Ishan; Gundawar, Manoj Kumar

2014-01-01

We demonstrate the application of non-gated laser induced breakdown spectroscopy (LIBS) for characterization and classification of organic materials with similar chemical composition. While use of such a system introduces substantive continuum background in the spectral dataset, we show that appropriate treatment of the continuum and characteristic emission results in accurate discrimination of pharmaceutical formulations of similar stoichiometry. Specifically, our results suggest that near-perfect classification can be obtained by employing suitable multivariate analysis on the acquired spectra, without prior removal of the continuum background. Indeed, we conjecture that pre-processing in the form of background removal may introduce spurious features in the signal. Our findings in this report significantly advance the prior results in time-integrated LIBS application and suggest the possibility of a portable, non-gated LIBS system as a process analytical tool, given its simple instrumentation needs, real-time capability and lack of sample preparation requirements. PMID:25084522

Food Forensics: Using Mass Spectrometry To Detect Foodborne Protein Contaminants, as Exemplified by Shiga Toxin Variants and Prion Strains.

PubMed

Silva, Christopher J

2018-06-13

Food forensicists need a variety of tools to detect the many possible food contaminants. As a result of its analytical flexibility, mass spectrometry is one of those tools. Use of the multiple reaction monitoring (MRM) method expands its use to quantitation as well as detection of infectious proteins (prions) and protein toxins, such as Shiga toxins. The sample processing steps inactivate prions and Shiga toxins; the proteins are digested with proteases to yield peptides suitable for MRM-based analysis. Prions are detected by their distinct physicochemical properties and differential covalent modification. Shiga toxin analysis is based on detecting peptides derived from the five identical binding B subunits comprising the toxin. 15 N-labeled internal standards are prepared from cloned proteins. These examples illustrate the power of MRM, in that the same instrument can be used to safely detect and quantitate protein toxins, prions, and small molecules that might contaminate our food.

[Recent advances in sample preparation methods of plant hormones].

PubMed

Wu, Qian; Wang, Lus; Wu, Dapeng; Duan, Chunfeng; Guan, Yafeng

2014-04-01

Plant hormones are a group of naturally occurring trace substances which play a crucial role in controlling the plant development, growth and environment response. With the development of the chromatography and mass spectroscopy technique, chromatographic analytical method has become a widely used way for plant hormone analysis. Among the steps of chromatographic analysis, sample preparation is undoubtedly the most vital one. Thus, a highly selective and efficient sample preparation method is critical for accurate identification and quantification of phytohormones. For the three major kinds of plant hormones including acidic plant hormones & basic plant hormones, brassinosteroids and plant polypeptides, the sample preparation methods are reviewed in sequence especially the recently developed methods. The review includes novel methods, devices, extractive materials and derivative reagents for sample preparation of phytohormones analysis. Especially, some related works of our group are included. At last, the future developments in this field are also prospected.

Identification of Particles in Parenteral Drug Raw Materials.

PubMed

Lee, Kathryn; Lankers, Markus; Valet, Oliver

2018-04-18

Particles in drug products are not good and are therefore regulated. These particles can come from the very beginning of the manufacturing process, from the raw materials. To prevent particles, it is important to understand what they are and where they come from so the raw material quality, processing, and shipping can be improved. Thus, it is important to correctly identify particles seen in raw materials. Raw materials need to be of a certain quality with respect to physical and chemical composition, and need to have no contaminants in the form of particles which could contaminate the product or indicate the raw materials are not pure enough to make a good quality product. Particles are often seen when handling raw materials due to color, size, or shape characteristics different from those in the raw materials. Particles may appear to the eye to be very different things than they actually are, so microscope, chemical, and elemental analyses are required for accuracy in proper identification. This paper shows how using three different spectroscopy tools correctly and together can be used to identify particles from extrinsic, intrinsic, and inherent particles. Sources of materials can be humans and the environment (extrinsic), from within the process (intrinsic), and part of the formulation (inherent). Microscope versions of Raman spectroscopy, laser-induced breakdown spectroscopy (LIBS), and IR spectroscopy are excellent tools for identifying particles because they are fast and accurate techniques needing minimal sample preparation that can provide chemical composition as well as images that can be used for identification. The micro analysis capabilities allow for easy analysis of different portions of samples so multiple components can be identified and sample preparation can be reduced. Using just one of these techniques may not be sufficient to give adequate identification results so that the source of contamination can be adequately identified. The complementarity of the techniques provides the advantage of identifying various chemical and molecular components, as well as elemental and image analyses. Correct interpretation of the results from these techniques is also very important. Copyright © 2018, Parenteral Drug Association.

Inverse supercritical fluid extraction as a sample preparation method for the analysis of the nanoparticle content in sunscreen agents.

PubMed

Müller, David; Cattaneo, Stefano; Meier, Florian; Welz, Roland; de Vries, Tjerk; Portugal-Cohen, Meital; Antonio, Diana C; Cascio, Claudia; Calzolai, Luigi; Gilliland, Douglas; de Mello, Andrew

2016-04-01

We demonstrate the use of inverse supercritical carbon dioxide (scCO2) extraction as a novel method of sample preparation for the analysis of complex nanoparticle-containing samples, in our case a model sunscreen agent with titanium dioxide nanoparticles. The sample was prepared for analysis in a simplified process using a lab scale supercritical fluid extraction system. The residual material was easily dispersed in an aqueous solution and analyzed by Asymmetrical Flow Field-Flow Fractionation (AF4) hyphenated with UV- and Multi-Angle Light Scattering detection. The obtained results allowed an unambiguous determination of the presence of nanoparticles within the sample, with almost no background from the matrix itself, and showed that the size distribution of the nanoparticles is essentially maintained. These results are especially relevant in view of recently introduced regulatory requirements concerning the labeling of nanoparticle-containing products. The novel sample preparation method is potentially applicable to commercial sunscreens or other emulsion-based cosmetic products and has important ecological advantages over currently used sample preparation techniques involving organic solvents. Copyright © 2016 Elsevier B.V. All rights reserved.

Sample preparation techniques for the determination of trace residues and contaminants in foods.

PubMed

Ridgway, Kathy; Lalljie, Sam P D; Smith, Roger M

2007-06-15

The determination of trace residues and contaminants in complex matrices, such as food, often requires extensive sample extraction and preparation prior to instrumental analysis. Sample preparation is often the bottleneck in analysis and there is a need to minimise the number of steps to reduce both time and sources of error. There is also a move towards more environmentally friendly techniques, which use less solvent and smaller sample sizes. Smaller sample size becomes important when dealing with real life problems, such as consumer complaints and alleged chemical contamination. Optimal sample preparation can reduce analysis time, sources of error, enhance sensitivity and enable unequivocal identification, confirmation and quantification. This review considers all aspects of sample preparation, covering general extraction techniques, such as Soxhlet and pressurised liquid extraction, microextraction techniques such as liquid phase microextraction (LPME) and more selective techniques, such as solid phase extraction (SPE), solid phase microextraction (SPME) and stir bar sorptive extraction (SBSE). The applicability of each technique in food analysis, particularly for the determination of trace organic contaminants in foods is discussed.

Sample Preparation Techniques for Grain Boundary Characterization of Annealed TRISO-Coated Particles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunzik-Gougar, M. L.; van Rooyen, I. J.; Hill, C. M.

Crystallographic information about chemical vapor deposition layers of silicon carbide (SiC) is essential to understanding layer performance, especially when the layers are in non planar geometries, such as spherical. We performed electron Back Scatter Diffraction (EBSD) analysis of spherical SiC layers using a different approach to sample focus ion beam milling technique to avoid the negative impacts of traditional sample polishing and to address the need of very small samples of irradiated materials for analysis. Mechanical and chemical grinding and polishing of sample surfaces can introduce lattice strains and result in unequal removal of SiC and surrounding layers of differentmore » material due to the hardness differences of these materials. The nature of layer interfaces is thought to play a key role in performance of the SiC; therefore, analysis of representative samples at these interfacial areas is crucial. In work reported here, a focused ion beam (FIB) was employed in a novel manner to prepare a more representative sample for EBSD analysis from TRISO layers free of effects introduced by mechanical and chemical preparation methods. In addition, the difficulty of handling neutron irradiated microscopic samples such as those analyzed in this work has been simplified with pre tilted mounting stages. Our study showed that although the average grain size of samples may be similar, the grain boundary characteristics may differ significantly. It was also found that low angle grain boundaries, comprises 25% in the FIB-prepared sample vs only 1-2% in the polished sample measured in the same particle. From this study it was determined that results of FIB prepared sample will provide more repeatable results, as the role of sample preparation is eliminated.« less

Sample Preparation Techniques for Grain Boundary Characterization of Annealed TRISO-Coated Particles

DOE PAGES

Dunzik-Gougar, M. L.; van Rooyen, I. J.; Hill, C. M.; ...

2016-08-25

Crystallographic information about chemical vapor deposition layers of silicon carbide (SiC) is essential to understanding layer performance, especially when the layers are in non planar geometries, such as spherical. We performed electron Back Scatter Diffraction (EBSD) analysis of spherical SiC layers using a different approach to sample focus ion beam milling technique to avoid the negative impacts of traditional sample polishing and to address the need of very small samples of irradiated materials for analysis. Mechanical and chemical grinding and polishing of sample surfaces can introduce lattice strains and result in unequal removal of SiC and surrounding layers of differentmore » material due to the hardness differences of these materials. The nature of layer interfaces is thought to play a key role in performance of the SiC; therefore, analysis of representative samples at these interfacial areas is crucial. In work reported here, a focused ion beam (FIB) was employed in a novel manner to prepare a more representative sample for EBSD analysis from TRISO layers free of effects introduced by mechanical and chemical preparation methods. In addition, the difficulty of handling neutron irradiated microscopic samples such as those analyzed in this work has been simplified with pre tilted mounting stages. Our study showed that although the average grain size of samples may be similar, the grain boundary characteristics may differ significantly. It was also found that low angle grain boundaries, comprises 25% in the FIB-prepared sample vs only 1-2% in the polished sample measured in the same particle. From this study it was determined that results of FIB prepared sample will provide more repeatable results, as the role of sample preparation is eliminated.« less

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

B. D. Miller; J. Gan; J. Madden

2012-05-01

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.

40 CFR 761.323 - Sample preparation.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Remediation Waste Samples § 761.323 Sample preparation. (a) The comparison study requires analysis of a minimum of 10 samples weighing at least 300 grams each. Samples of PCB remediation waste used in the... PCB remediation waste at the cleanup site, or must be the same kind of material as that waste. For...

Proteoglycomics: Recent Progress and Future Challenges

PubMed Central

Ly, Mellisa; Laremore, Tatiana N.

2010-01-01

Abstract Proteoglycomics is a systematic study of structure, expression, and function of proteoglycans, a posttranslationally modified subset of a proteome. Although relying on the established technologies of proteomics and glycomics, proteoglycomics research requires unique approaches for elucidating structure–function relationships of both proteoglycan components, glycosaminoglycan chain, and core protein. This review discusses our current understanding of structure and function of proteoglycans, major players in the development, normal physiology, and disease. A brief outline of the proteoglycomic sample preparation and analysis is provided along with examples of several recent proteoglycomic studies. Unique challenges in the characterization of glycosaminoglycan component of proteoglycans are discussed, with emphasis on the many analytical tools used and the types of information they provide. PMID:20450439

Modular microfluidic system for biological sample preparation

DOEpatents

Rose, Klint A.; Mariella, Jr., Raymond P.; Bailey, Christopher G.; Ness, Kevin Dean

2015-09-29

A reconfigurable modular microfluidic system for preparation of a biological sample including a series of reconfigurable modules for automated sample preparation adapted to selectively include a) a microfluidic acoustic focusing filter module, b) a dielectrophoresis bacteria filter module, c) a dielectrophoresis virus filter module, d) an isotachophoresis nucleic acid filter module, e) a lyses module, and f) an isotachophoresis-based nucleic acid filter.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wen, Haiming; Lin, Yaojun; Seidman, David N.

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 dimpledmore » 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.« less

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

DOE PAGES

Wen, Haiming; Lin, Yaojun; Seidman, David N.; ...

2015-09-09

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 dimpledmore » 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.« less

Critical evaluation of sample pretreatment techniques.

PubMed

Hyötyläinen, Tuulia

2009-06-01

Sample preparation before chromatographic separation is the most time-consuming and error-prone part of the analytical procedure. Therefore, selecting and optimizing an appropriate sample preparation scheme is a key factor in the final success of the analysis, and the judicious choice of an appropriate procedure greatly influences the reliability and accuracy of a given analysis. The main objective of this review is to critically evaluate the applicability, disadvantages, and advantages of various sample preparation techniques. Particular emphasis is placed on extraction techniques suitable for both liquid and solid samples.

The Biobanking Analysis Resource Catalogue (BARCdb): a new research tool for the analysis of biobank samples

PubMed Central

Galli, Joakim; Oelrich, Johan; Taussig, Michael J.; Andreasson, Ulrika; Ortega-Paino, Eva; Landegren, Ulf

2015-01-01

We report the development of a new database of technology services and products for analysis of biobank samples in biomedical research. BARCdb, the Biobanking Analysis Resource Catalogue (http://www.barcdb.org), is a freely available web resource, listing expertise and molecular resource capabilities of research centres and biotechnology companies. The database is designed for researchers who require information on how to make best use of valuable biospecimens from biobanks and other sample collections, focusing on the choice of analytical techniques and the demands they make on the type of samples, pre-analytical sample preparation and amounts needed. BARCdb has been developed as part of the Swedish biobanking infrastructure (BBMRI.se), but now welcomes submissions from service providers throughout Europe. BARCdb can help match resource providers with potential users, stimulating transnational collaborations and ensuring compatibility of results from different labs. It can promote a more optimal use of European resources in general, both with respect to standard and more experimental technologies, as well as for valuable biobank samples. This article describes how information on service and reagent providers of relevant technologies is made available on BARCdb, and how this resource may contribute to strengthening biomedical research in academia and in the biotechnology and pharmaceutical industries. PMID:25336620

Acid-etching technique of non-decalcified bone samples for visualizing osteocyte-lacuno-canalicular network using scanning electron microscope.

PubMed

Lampi, Tiina; Dekker, Hannah; Ten Bruggenkate, Chris M; Schulten, Engelbert A J M; Mikkonen, Jopi J W; Koistinen, Arto; Kullaa, Arja M

2018-01-01

The aim of this study was to define the acid-etching technique for bone samples embedded in polymethyl metacrylate (PMMA) in order to visualize the osteocyte lacuno-canalicular network (LCN) for scanning electron microscopy (SEM). Human jaw bone tissue samples (N = 18) were collected from the study population consisting of patients having received dental implant surgery. After collection, the bone samples were fixed in 70% ethanol and non-decalcified samples embedded routinely into polymethyl metacrylate (PMMA). The PMMA embedded specimens were acid-etched in either 9 or 37% phosphoric acid (PA) and prepared for SEM for further analysis. PMMA embedded bone specimens acid-etched by 9% PA concentration accomplishes the most informative and favorable visualization of the LCN to be observed by SEM. Etching of PMMA embedded specimens is recommendable to start with 30 s or 40 s etching duration in order to find the proper etching duration for the samples examined. Visualizing osteocytes and LCN provides a tool to study bone structure that reflects changes in bone metabolism and diseases related to bone tissue. By proper etching protocol of non-decalcified and using scanning electron microscope it is possible to visualize the morphology of osteocytes and the network supporting vitality of bone tissue.

The OSIRIS-Rex Asteroid Sample Return: Mission Operations Design

NASA Technical Reports Server (NTRS)

Gal-Edd, Jonathan; Cheuvront, Allan

2014-01-01

The OSIRIS-REx mission employs a methodical, phased approach to ensure success in meeting the missions science requirements. OSIRIS-REx launches in September 2016, with a backup launch period occurring one year later. Sampling occurs in 2019. The departure burn from Bennu occurs in March 2021. On September 24, 2023, the SRC lands at the Utah Test and Training Range (UTTR). Stardust heritage procedures are followed to transport the SRC to Johnson Space Center, where the samples are removed and delivered to the OSIRIS-REx curation facility. After a six-month preliminary examination period the mission will produce a catalog of the returned sample, allowing the worldwide community to request samples for detailed analysis.Traveling and returning a sample from an Asteroid that has not been explored before requires unique operations consideration. The Design Reference Mission (DRM) ties together space craft, instrument and operations scenarios. The project implemented lessons learned from other small body missions: APLNEAR, JPLDAWN and ESARosetta. The key lesson learned was expected the unexpected and implement planning tools early in the lifecycle. In preparation to PDR, the project changed the asteroid arrival date, to arrive one year earlier and provided additional time margin. STK is used for Mission Design and STKScheduler for instrument coverage analysis.

Pre-analytical issues in the haemostasis laboratory: guidance for the clinical laboratories.

PubMed

Magnette, A; Chatelain, M; Chatelain, B; Ten Cate, H; Mullier, F

2016-01-01

Ensuring quality has become a daily requirement in laboratories. In haemostasis, even more than in other disciplines of biology, quality is determined by a pre-analytical step that encompasses all procedures, starting with the formulation of the medical question, and includes patient preparation, sample collection, handling, transportation, processing, and storage until time of analysis. This step, based on a variety of manual activities, is the most vulnerable part of the total testing process and is a major component of the reliability and validity of results in haemostasis and constitutes the most important source of erroneous or un-interpretable results. Pre-analytical errors may occur throughout the testing process and arise from unsuitable, inappropriate or wrongly handled procedures. Problems may arise during the collection of blood specimens such as misidentification of the sample, use of inadequate devices or needles, incorrect order of draw, prolonged tourniquet placing, unsuccessful attempts to locate the vein, incorrect use of additive tubes, collection of unsuitable samples for quality or quantity, inappropriate mixing of a sample, etc. Some factors can alter the result of a sample constituent after collection during transportation, preparation and storage. Laboratory errors can often have serious adverse consequences. Lack of standardized procedures for sample collection accounts for most of the errors encountered within the total testing process. They can also have clinical consequences as well as a significant impact on patient care, especially those related to specialized tests as these are often considered as "diagnostic". Controlling pre-analytical variables is critical since this has a direct influence on the quality of results and on their clinical reliability. The accurate standardization of the pre-analytical phase is of pivotal importance for achieving reliable results of coagulation tests and should reduce the side effects of the influence factors. This review is a summary of the most important recommendations regarding the importance of pre-analytical factors for coagulation testing and should be a tool to increase awareness about the importance of pre-analytical factors for coagulation testing.

The archaeal diversity in a cave system and its implications for life on other planets

NASA Astrophysics Data System (ADS)

Leuko, Stefan; Rettberg, Petra; De Waele, Jo; Bessone, Loredana; Sauro, Francesco; Sanna, Laura

The quest of exploring and looking for life in new places is a human desire since centuries. Nowadays, we are not only looking on planet Earth any more, but our endeavours focus on nearby planets in our solar system. At this point in time, we are not able to send manned missions to other planets, but to be ready and prepared for the day, training today is pivotal. Developed by the European Space Agency (ESA) since 2008, these CAVES missions (Cooperative Adventure for Valuing and Exercising human behaviour and performance Skills), prepare astronauts to work safely and effectively and solve problems as a multicultural team while exploring uncharted underground natural areas (i.e. caves) using space procedures. The hypogean environment is also of great interest for astrobiological research as cave conditions may resemble those in extra-terrestrial environments. Besides the main focus of exploration and skill training, future astronauts are also trained in taking microbiological samples for analysis during the exploration and for further analysis in the lab. During the 2013 mission, astronauts collected soil samples and employed flocked swaps to sample areas with little or no visible soil. Microscopic analysis back in the lab revealed a diverse spectrum of different cell shapes and sizes. Samples were further analysed employing molecular tools such as RFLP analysis, 16s rRNA clone libraries and sequence analysis. RFLP pattern analysis revealed that the community can be divided in 9 main groups and several single patterns. The largest group (40% of all analysed clones) belong to the clade of ammonia oxidizing archaea Nitrosopumilus spp.. Dividing the results by sampling point, it showed that the highest diversity of organisms was located on the flocked swaps, which is interesting as the sample was taken from a rock surface with no visible organic matter. By analysis low energy systems like a cave, we may be able to find clues for what could be necessary to survive on a different planet.

Federal agency transportation management program handbook

DOT National Transportation Integrated Search

1999-04-01

This handbook provides Federal agencies in the National Capital Region (NCR) with procedures and guidelines on how to prepare a Transportation Management Program (TMP). The preparation of TMPs is one tool in the overall field of Travel Demand Managem...

Preparation of protein samples for mass spectrometry and N-terminal sequencing.

PubMed

Glenn, Gary

2014-01-01

The preparation of protein samples for mass spectrometry and N-terminal sequencing is a key step in successfully identifying proteins. Mass spectrometry is a very sensitive technique, and as such, samples must be prepared carefully since they can be subject to contamination of the sample (e.g., due to incomplete subcellular fractionation or purification of a multiprotein complex), overwhelming of the sample by highly abundant proteins, and contamination from skin or hair (keratin can be a very common hit). One goal of sample preparation for mass spec is to reduce the complexity of the sample - in the example presented here, mitochondria are purified, solubilized, and fractionated by sucrose density gradient sedimentation prior to preparative 1D SDS-PAGE. It is important to verify the purity and integrity of the sample so that you can have confidence in the hits obtained. More protein is needed for N-terminal sequencing and ideally it should be purified to a single band when run on an SDS-polyacrylamide gel. The example presented here involves stably expressing a tagged protein in HEK293 cells and then isolating the protein by affinity purification and SDS-PAGE. © 2014 Elsevier Inc. All rights reserved.

78 FR 45265 - Notice of Intent To Prepare a Resource Management Plan Amendment and Associated Environmental...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-26

... Environmental Assessment (EA) to consider leasing Federal coal in response to lease application ALES-55199 for.... A mine plan scenario will be prepared for the Federal coal resource as an analytical tool to inform... Prepare a Resource Management Plan Amendment and Associated Environmental Assessment for Coal Lease by...

Preparation and characterizations of activated carbon monolith from rubber wood and its effect on supercapacitor performances

NASA Astrophysics Data System (ADS)

Taer, E.; Taslim, R.; Deraman, M.

2016-02-01

Preparation of activated carbon monolith (ACM) from rubber wood was investigated. Two kind of preparation method were carried out by pre-carbonized of rubber wood saw dust and rubber wood material as it is naturally. The samples were prepared with pelletizing method and small cutting of rubber wood in cross sectional method. Both of samples were characterized by physical and electrochemical technique. The physical properties such as morphology and porosity were investigated. The electrochemical properties of both samples such as equivalent series resistances (ESR) and specific capacitances were also compared. In conclusion, this study showed that both of different preparation method would propose a simple method of ACM electrode preparation technique for supercapacitor applications.

A high-throughput robotic sample preparation system and HPLC-MS/MS for measuring urinary anatabine, anabasine, nicotine and major nicotine metabolites.

PubMed

Wei, Binnian; Feng, June; Rehmani, Imran J; Miller, Sharyn; McGuffey, James E; Blount, Benjamin C; Wang, Lanqing

2014-09-25

Most sample preparation methods characteristically involve intensive and repetitive labor, which is inefficient when preparing large numbers of samples from population-scale studies. This study presents a robotic system designed to meet the sampling requirements for large population-scale studies. Using this robotic system, we developed and validated a method to simultaneously measure urinary anatabine, anabasine, nicotine and seven major nicotine metabolites: 4-Hydroxy-4-(3-pyridyl)butanoic acid, cotinine-N-oxide, nicotine-N-oxide, trans-3'-hydroxycotinine, norcotinine, cotinine and nornicotine. We analyzed robotically prepared samples using high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometry in positive electrospray ionization mode using scheduled multiple reaction monitoring (sMRM) with a total runtime of 8.5 min. The optimized procedure was able to deliver linear analyte responses over a broad range of concentrations. Responses of urine-based calibrators delivered coefficients of determination (R(2)) of >0.995. Sample preparation recovery was generally higher than 80%. The robotic system was able to prepare four 96-well plate (384 urine samples) per day, and the overall method afforded an accuracy range of 92-115%, and an imprecision of <15.0% on average. The validation results demonstrate that the method is accurate, precise, sensitive, robust, and most significantly labor-saving for sample preparation, making it efficient and practical for routine measurements in large population-scale studies such as the National Health and Nutrition Examination Survey (NHANES) and the Population Assessment of Tobacco and Health (PATH) study. Published by Elsevier B.V.

Deriving Earth Science Data Analytics Tools/Techniques Requirements

NASA Astrophysics Data System (ADS)

Kempler, S. J.

2015-12-01

Data Analytics applications have made successful strides in the business world where co-analyzing extremely large sets of independent variables have proven profitable. Today, most data analytics tools and techniques, sometimes applicable to Earth science, have targeted the business industry. In fact, the literature is nearly absent of discussion about Earth science data analytics. Earth science data analytics (ESDA) is the process of examining large amounts of data from a variety of sources to uncover hidden patterns, unknown correlations, and other useful information. ESDA is most often applied to data preparation, data reduction, and data analysis. Co-analysis of increasing number and volume of Earth science data has become more prevalent ushered by the plethora of Earth science data sources generated by US programs, international programs, field experiments, ground stations, and citizen scientists. Through work associated with the Earth Science Information Partners (ESIP) Federation, ESDA types have been defined in terms of data analytics end goals. Goals of which are very different than those in business, requiring different tools and techniques. A sampling of use cases have been collected and analyzed in terms of data analytics end goal types, volume, specialized processing, and other attributes. The goal of collecting these use cases is to be able to better understand and specify requirements for data analytics tools and techniques yet to be implemented. This presentation will describe the attributes and preliminary findings of ESDA use cases, as well as provide early analysis of data analytics tools/techniques requirements that would support specific ESDA type goals. Representative existing data analytics tools/techniques relevant to ESDA will also be addressed.

Studies of tin-transition metal-carbon and tin-cobalt-transition metal-carbon negative electrode materials prepared by mechanical attrition

NASA Astrophysics Data System (ADS)

Ferguson, P. P.; Martine, M. L.; George, A. E.; Dahn, J. R.

Samples of Sn 30TM 30C 40 and of Sn 30Co 15TM 15C 40, with TM = 3d transition metals, were prepared by vertical-axis attritor milling. The structure and performance of these samples were studied by X-ray diffraction (XRD) and by electrochemical testing. The XRD patterns of Sn 30TM 30C 40 show an amorphous-like diffraction pattern only for the sample with TM = Co. The other prepared samples show broadened Bragg peaks of their main starting material, along with an amorphous-like background, even after 32 h of milling. Samples with TM = Co and TM = Ni show stable differential capacity versus potential plots and stable cycling for at least 100 cycles with reversible capacities of 425 and 250 mAh g -1, respectively. All samples prepared with 15 at.% Co show good capacity retention for at least 100 cycles ranging from 270 mAh g -1 for samples with TM = Ni to 500 mAh g -1 for samples with TM = Ti. The differential capacity versus potential plots for all the prepared Sn 30Co 15TM 15C 40 samples show similar structure to that of Sn 30Co 30C 40 except when TM = Cu. This shows the possibility of preparing tin-based negative electrode materials using a combination of cobalt and TM, especially if one looks to reduce the cobalt content.

Non-Contact Conductivity Measurement for Automated Sample Processing Systems

NASA Technical Reports Server (NTRS)

Beegle, Luther W.; Kirby, James P.

2012-01-01

A new method has been developed for monitoring and control of automated sample processing and preparation especially focusing on desalting of samples before analytical analysis (described in more detail in Automated Desalting Apparatus, (NPO-45428), NASA Tech Briefs, Vol. 34, No. 8 (August 2010), page 44). The use of non-contact conductivity probes, one at the inlet and one at the outlet of the solid phase sample preparation media, allows monitoring of the process, and acts as a trigger for the start of the next step in the sequence (see figure). At each step of the muti-step process, the system is flushed with low-conductivity water, which sets the system back to an overall low-conductivity state. This measurement then triggers the next stage of sample processing protocols, and greatly minimizes use of consumables. In the case of amino acid sample preparation for desalting, the conductivity measurement will define three key conditions for the sample preparation process. First, when the system is neutralized (low conductivity, by washing with excess de-ionized water); second, when the system is acidified, by washing with a strong acid (high conductivity); and third, when the system is at a basic condition of high pH (high conductivity). Taken together, this non-contact conductivity measurement for monitoring sample preparation will not only facilitate automation of the sample preparation and processing, but will also act as a way to optimize the operational time and use of consumables

Quantification of microscopic surface features of single point diamond turned optics with subsequent chemical polishing

NASA Astrophysics Data System (ADS)

Cardenas, Nelson; Kyrish, Matthew; Taylor, Daniel; Fraelich, Margaret; Lechuga, Oscar; Claytor, Richard; Claytor, Nelson

2015-03-01

Electro-Chemical Polishing is routinely used in the anodizing industry to achieve specular surface finishes of various metals products prior to anodizing. Electro-Chemical polishing functions by leveling the microscopic peaks and valleys of the substrate, thereby increasing specularity and reducing light scattering. The rate of attack is dependent of the physical characteristics (height, depth, and width) of the microscopic structures that constitute the surface finish. To prepare the sample, mechanical polishing such as buffing or grinding is typically required before etching. This type of mechanical polishing produces random microscopic structures at varying depths and widths, thus the electropolishing parameters are determined in an ad hoc basis. Alternatively, single point diamond turning offers excellent repeatability and highly specific control of substrate polishing parameters. While polishing, the diamond tool leaves behind an associated tool mark, which is related to the diamond tool geometry and machining parameters. Machine parameters such as tool cutting depth, speed and step over can be changed in situ, thus providing control of the spatial frequency of the microscopic structures characteristic of the surface topography of the substrate. By combining single point diamond turning with subsequent electro-chemical etching, ultra smooth polishing of both rotationally symmetric and free form mirrors and molds is possible. Additionally, machining parameters can be set to optimize post polishing for increased surface quality and reduced processing times. In this work, we present a study of substrate surface finish based on diamond turning tool mark spatial frequency with subsequent electro-chemical polishing.

On-machine precision preparation and dressing of ball-headed diamond wheel for the grinding of fused silica

NASA Astrophysics Data System (ADS)

Chen, Mingjun; Li, Ziang; Yu, Bo; Peng, Hui; Fang, Zhen

2013-09-01

In the grinding of high quality fused silica parts with complex surface or structure using ball-headed metal bonded diamond wheel with small diameter, the existing dressing methods are not suitable to dress the ball-headed diamond wheel precisely due to that they are either on-line in process dressing which may causes collision problem or without consideration for the effects of the tool setting error and electrode wear. An on-machine precision preparation and dressing method is proposed for ball-headed diamond wheel based on electrical discharge machining. By using this method the cylindrical diamond wheel with small diameter is manufactured to hemispherical-headed form. The obtained ball-headed diamond wheel is dressed after several grinding passes to recover geometrical accuracy and sharpness which is lost due to the wheel wear. A tool setting method based on high precision optical system is presented to reduce the wheel center setting error and dimension error. The effect of electrode tool wear is investigated by electrical dressing experiments, and the electrode tool wear compensation model is established based on the experimental results which show that the value of wear ratio coefficient K' tends to be constant with the increasing of the feed length of electrode and the mean value of K' is 0.156. Grinding experiments of fused silica are carried out on a test bench to evaluate the performance of the preparation and dressing method. The experimental results show that the surface roughness of the finished workpiece is 0.03 μm. The effect of the grinding parameter and dressing frequency on the surface roughness is investigated based on the measurement results of the surface roughness. This research provides an on-machine preparation and dressing method for ball-headed metal bonded diamond wheel used in the grinding of fused silica, which provides a solution to the tool setting method and the effect of electrode tool wear.

Cost Analysis of Various Low Pathogenic Avian Influenza Surveillance Systems in the Dutch Egg Layer Sector

PubMed Central

Rutten, Niels; Gonzales, José L.; Elbers, Armin R. W.; Velthuis, Annet G. J.

2012-01-01

Background As low pathogenic avian influenza viruses can mutate into high pathogenic viruses the Dutch poultry sector implemented a surveillance system for low pathogenic avian influenza (LPAI) based on blood samples. It has been suggested that egg yolk samples could be sampled instead of blood samples to survey egg layer farms. To support future decision making about AI surveillance economic criteria are important. Therefore a cost analysis is performed on systems that use either blood or eggs as sampled material. Methodology/Principal Findings The effectiveness of surveillance using egg or blood samples was evaluated using scenario tree models. Then an economic model was developed that calculates the total costs for eight surveillance systems that have equal effectiveness. The model considers costs for sampling, sample preparation, sample transport, testing, communication of test results and for the confirmation test on false positive results. The surveillance systems varied in sampled material (eggs or blood), sampling location (farm or packing station) and location of sample preparation (laboratory or packing station). It is shown that a hypothetical system in which eggs are sampled at the packing station and samples prepared in a laboratory had the lowest total costs (i.e. € 273,393) a year. Compared to this a hypothetical system in which eggs are sampled at the farm and samples prepared at a laboratory, and the currently implemented system in which blood is sampled at the farm and samples prepared at a laboratory have 6% and 39% higher costs respectively. Conclusions/Significance This study shows that surveillance for avian influenza on egg yolk samples can be done at lower costs than surveillance based on blood samples. The model can be used in future comparison of surveillance systems for different pathogens and hazards. PMID:22523543

Tools and Their Uses. Rate Training Manual.

ERIC Educational Resources Information Center

Naval Personnel Program Support Activity, Washington, DC.

One of a series of training manuals prepared for enlisted personnel in the Navy and Naval Reserve, this supplementary manual contains data pertinent to a variety of tools necessary to the satisfactory performance of modern technical equipment used by the Navy. It is designed to help the learner identify tools and fastening devices by their correct…

Mobilization Protocols for Hybrid Sensors for Environmental AOP Sampling (HySEAS) Observations

NASA Technical Reports Server (NTRS)

Hooker, Stanford B.

2014-01-01

The protocols presented here enable the proper mobilization of the latest-generation instruments for measuring the apparent optical properties (AOPs) of aquatic ecosystems. The protocols are designed for the Hybrid Sensors for Environmental AOP Sampling (HySEAS) class of instruments, but are applicable to the community of practice for AOP measurements. The protocols are organized into eleven sections beyond an introductory overview: a) cables and connectors, b) HySEAS instruments, c) platform preparation, d) instrument installation, e) cable installation, f) test deployment, g) test recovery, h) maintenance, i) shipping, j) storage, and k) smallboat operations. Each section concentrates on documenting how to prevent the most likely faults, remedy them should they occur, and accomplishing both with the proper application of a modest set of useful tools. Within the twelve sections, there are Socratic exercises to stimulate thought, and the answers to these exercises appear in Appendix A. Frequently asked questions (FAQs) are summarized in a separate section after the answers to the exercises in Appendix B. For practitioners unfamiliar with the nautical terms used throughout this document plus others likely encountered at sea, an abbreviated dictionary of nautical terms appears in Appendix C. An abbreviated dictionary of radiotelephone terms is presented in Appendix D. To ensure familiarity with many of the tools that are presented, Appendix E provides a description of the tools alongside a thumbnail picture. Abbreviated deployment checklists and cable diagrams are provided in Appendix F. The document concludes with an acknowledgments section, a glossary of acronyms, a definition of symbols, and a list of references.

An REU Project on the Precambrian Rocks of Yellowstone National Park: Some lessons learned

NASA Astrophysics Data System (ADS)

Henry, D.; Mogk, D. W.; Mueller, P. A.; Foster, D. A.

2014-12-01

An NSF-funded REU project (2011-2013), based in Yellowstone National Park (YNP), was designed to characterize the geology, geochemistry and geochronology of Precambrian rocks in northern YNP. Over two field seasons two cadres of 12 students (12 women and 12 men) were chosen from small-to-large state universities and private colleges. REU students participated in three major activities constituting a complete research experience: Field studies involved geologic mapping and sampling of Precambrian basement; formulation of testable research questions by smaller working groups; and mapping and sampling projects to address research questions; Analytical studies, sample preparation immediately followed field work with petrographic analysis at students' home institutions and a week-long visit to analytical laboratories to conduct follow-up studies by small research groups during the academic year (Univ. Florida - geochemistry and geochronology; Univ. Minnesota - EMPA analysis); Communicating results, each working group submitted an abstract and collectively presented 13 posters at the 2011 and 2012 GSA Rocky Mountain sectional meetings. We used directed discovery to engage students in a community of practice in the field and found that a long apprenticeship (2-3 weeks) is optimal for novice-master interactions in exploring natural setting. Initial group hikes were used to normalize methods and language of the discipline. Students developed a sense of ownership of the overall project and assumed personal responsibility for directed research projects. Training was provided to: guide students in selection and appropriate use of tools; develop sampling strategies; discuss communal ethics, values, and expectations; develop efficient work habits; stimulate independent thinking; and engage decision-making. It was important to scaffold the field experience to students' level of development to lead to mastery. Analytical activities were designed from rock to analysis so that each group mastered all preparation steps and instrumental techniques under supervision of graduate mentors and lab managers leading to a clearer understanding of data interpretation. Students were communally engaged in abstract and poster preparation to ensure proper focus, scientific breadth, and style of presentation.

An automation-assisted generic approach for biological sample preparation and LC-MS/MS method validation.

PubMed

Zhang, Jie; Wei, Shimin; Ayres, David W; Smith, Harold T; Tse, Francis L S

2011-09-01

Although it is well known that automation can provide significant improvement in the efficiency of biological sample preparation in quantitative LC-MS/MS analysis, it has not been widely implemented in bioanalytical laboratories throughout the industry. This can be attributed to the lack of a sound strategy and practical procedures in working with robotic liquid-handling systems. Several comprehensive automation assisted procedures for biological sample preparation and method validation were developed and qualified using two types of Hamilton Microlab liquid-handling robots. The procedures developed were generic, user-friendly and covered the majority of steps involved in routine sample preparation and method validation. Generic automation procedures were established as a practical approach to widely implement automation into the routine bioanalysis of samples in support of drug-development programs.

Thermoluminescence property of nano scale Al{sub 2}O{sub 3}: C by combustion method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bharthasaradhi, R.; Nehru, L. C.

In this study, thermoluminescence dosimetry material of carbon doped aluminium oxide by combustion method using Aluminium nitrate and Glycine. The Structure of the prepared Sample was carried out by XRD. The sample was nano crystalline in nature. Having hexagonal structure with unit cell parameters a=4.75Å, C=12.99Å. The surface morphology of the prepared nanopowder was carried out through (SEM). The morphology of the prepared sample is platelet structure and functional group analysis carried out through FT-IR Spectrum. The prepared sample was irradiated through γ-ray CO{sup 60} (100 Gy) was used as γ-ray source. The thermoluminescence glow curve of the irradiated samplemore » showed an isolated peak at around 200°C. The result suggest the prepared nanopowder is suitable for medical radiation dosimetry.« less

Automated digital image analysis of islet cell mass using Nikon's inverted eclipse Ti microscope and software to improve engraftment may help to advance the therapeutic efficacy and accessibility of islet transplantation across centers.

PubMed

Gmyr, Valery; Bonner, Caroline; Lukowiak, Bruno; Pawlowski, Valerie; Dellaleau, Nathalie; Belaich, Sandrine; Aluka, Isanga; Moermann, Ericka; Thevenet, Julien; Ezzouaoui, Rimed; Queniat, Gurvan; Pattou, Francois; Kerr-Conte, Julie

2015-01-01

Reliable assessment of islet viability, mass, and purity must be met prior to transplanting an islet preparation into patients with type 1 diabetes. The standard method for quantifying human islet preparations is by direct microscopic analysis of dithizone-stained islet samples, but this technique may be susceptible to inter-/intraobserver variability, which may induce false positive/negative islet counts. Here we describe a simple, reliable, automated digital image analysis (ADIA) technique for accurately quantifying islets into total islet number, islet equivalent number (IEQ), and islet purity before islet transplantation. Islets were isolated and purified from n = 42 human pancreata according to the automated method of Ricordi et al. For each preparation, three islet samples were stained with dithizone and expressed as IEQ number. Islets were analyzed manually by microscopy or automatically quantified using Nikon's inverted Eclipse Ti microscope with built-in NIS-Elements Advanced Research (AR) software. The AIDA method significantly enhanced the number of islet preparations eligible for engraftment compared to the standard manual method (p < 0.001). Comparisons of individual methods showed good correlations between mean values of IEQ number (r(2) = 0.91) and total islet number (r(2) = 0.88) and thus increased to r(2) = 0.93 when islet surface area was estimated comparatively with IEQ number. The ADIA method showed very high intraobserver reproducibility compared to the standard manual method (p < 0.001). However, islet purity was routinely estimated as significantly higher with the manual method versus the ADIA method (p < 0.001). The ADIA method also detected small islets between 10 and 50 µm in size. Automated digital image analysis utilizing the Nikon Instruments software is an unbiased, simple, and reliable teaching tool to comprehensively assess the individual size of each islet cell preparation prior to transplantation. Implementation of this technology to improve engraftment may help to advance the therapeutic efficacy and accessibility of islet transplantation across centers.

Final Report for X-ray Diffraction Sample Preparation Method Development

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ely, T. M.; Meznarich, H. K.; Valero, T.

WRPS-1500790, “X-ray Diffraction Saltcake Sample Preparation Method Development Plan/Procedure,” was originally prepared with the intent of improving the specimen preparation methodology used to generate saltcake specimens suitable for XRD-based solid phase characterization. At the time that this test plan document was originally developed, packed powder in cavity supports with collodion binder was the established XRD specimen preparation method. An alternate specimen preparation method less vulnerable, if not completely invulnerable to preferred orientation effects, was desired as a replacement for the method.

Fabricated CeO2 nanopowders as a novel sensing platform for advanced forensic, electrochemical and photocatalytic applications

NASA Astrophysics Data System (ADS)

Rohini, B. S.; Nagabhushana, H.; Darshan, G. P.; Basavaraj, R. B.; Sharma, S. C.; Sudarmani, R.

2017-11-01

In Forensic investigation, identification of various types of ridge details are essential in order to fix the criminals associated in various crimes. Even though several methods and labeling agents are available to visualize latent finger prints (LFPs) there is still simple, accurate, cost-effective, and non-destructive tool is required. In the present work, CeO2 nanopowders (NPs) are prepared via simple solution combustion route using Tamarindus indica fruit extract as a fuel. The optimized NPs are utilized for visualization of LFPs on various surfaces by powder dusting method. Results revealed that visualized LFPs exhibit Level 3 features such as pores and ridge contours under normal light with high sensitivity and without background hindrance. The photometric characteristics of the prepared samples exhibit blue color emission and highly useful in warm light emitting diodes. The photocatalytic studies were carried out with different Methylene blue (MB) dye concentration and pH values. The obtained results reveal that the CeO2 NPs exhibits an excellent catalytic properties which can act as a good catalytic reagent. The findings demonstrate that the prepared NPs are quite useful as a labeling agent for visualization of LFPs, efficient catalysts for dye degradation as well as solid-state lighting applications.

Pt decorated MoS2 nanoflakes for ultrasensitive resistive humidity sensor

NASA Astrophysics Data System (ADS)

Burman, Debasree; Santra, Sumita; Pramanik, Panchanan; Guha, Prasanta Kumar

2018-03-01

In this work, we report the fabrication of a low power, humidity sensor where platinum nanoparticles (NPs) decorated few-layered molybdenum disulphide (MoS2) nanoflakes have been used as the sensing layer. A mixed solvent was used to exfoliate the nanoflakes from the bulk powder. Then the Pt/MoS2 composites were prepared by reducing Pt NPs from chloroplatinic acid hexahydrate using a novel reduction technique using sulphide salt. The successful reduction and composite preparation were confirmed using various material characterization tools like scanning electron microscopy, atomic force microscopy, transmission electron microscopy, x-ray diffraction, x-ray photoelectron spectroscopy, Raman spectroscopy and UV-visible spectroscopy. The humidity sensors were prepared by drop-coating the Pt-decorated MoS2 on gold interdigitated electrodes and then exposed to various levels of relative humidity (RH). Composites with different weight ratios of Pt were tested and the best response was shown by the Pt/MoS2 (0.25:1) sample with a record high response of ˜4000 times at 85% RH. The response and recovery times were ˜92 s and ˜154 s respectively with repeatable behaviour. The sensor performance was found to be stable when tested over a few months. The underlying sensing mechanisms along with detailed characterization of the various composites have been discussed.

Prediction of recrystallization behavior of troglitazone/polyvinylpyrrolidone solid dispersion by solid-state NMR.

PubMed

Ito, Atsutoshi; Watanabe, Tomoyuki; Yada, Shuichi; Hamaura, Takeshi; Nakagami, Hiroaki; Higashi, Kenjirou; Moribe, Kunikazu; Yamamoto, Keiji

2010-01-04

The purpose of this study was to elaborate the relationship between the (13)C CP/MAS NMR spectra and the recrystallization behavior during the storage of troglitazone solid dispersions. The solid dispersions were prepared by either the solvent method or by co-grinding. The recrystallization behavior under storage conditions at 40 degrees C/94% RH was evaluated by the Kolmogorov-Johnson-Mehl-Avrami (KJMA) equation. Solid dispersions prepared by the solvent method or by prolonged grinding brought about inhibition of the nucleation and the nuclei growth at the same time. No differences in the PXRD profiles were found in the samples prepared by the co-grinding and solvent methods, however, (13)C CP/MAS NMR showed significant differences in the spectra. The correlation coefficients using partial least square regression analysis between the PXRD profiles and the apparent nuclei-growth constant or induction period to nucleation were 0.1305 or 0.6350, respectively. In contrast, those between the (13)C CP/MAS NMR spectra and the constant or the period were 0.9916 or 0.9838, respectively. The (13)C CP/MAS NMR spectra had good correlation with the recrystallization kinetic parameters evaluated by the KJMA equation. Consequently, solid-state NMR was judged to be a useful tool for the prediction of the recrystallization behavior of solid dispersions.

Quality assessment of compost prepared with municipal solid waste

NASA Astrophysics Data System (ADS)

Jodar, J. R.; Ramos, N.; Carreira, J. A.; Pacheco, R.; Fernández-Hernández, A.

2017-11-01

One way that helps maintain the sustainability of agro-ecosystems land is the application of compost from municipal solid waste as fertilizer, because it can recover the nutrients contained in them, minimizing the negative impact on the environment. Composting as a method for preparing organic fertilizers and amendments is economically and ecologically sound and may well represent an acceptable solution for disposing of municipal solid waste. In the present work, the quality of compost is studied made from municipal solid waste; the content of mineral nutrients: potassium, calcium, magnesium, sodium, zinc, manganese, cupper, iron, nickel, chromium and lead has been investigated. The objective was to evaluate the changes in mineral nutrient concentration during the composting process. The compost was prepared in a pilot-plant using the turning-pile system. Temperature was used as a monitoring parameter to follow the composting progress, which underwent the typical trend of municipal solid waste composting mixtures. The results showed a similar evolution on the content of mineral nutrients of the mixture of municipal solid waste. This evolution originated in a mature compost (end sample) with an adequate content of mineral elements and physical-chemical characteristics for its use in agriculture. So, the use of compost of municipal solid waste represents an important tool for fertilization requirements for its use in agriculture.

Acoustic Enrichment of Extracellular Vesicles from Biological Fluids.

PubMed

Ku, Anson; Lim, Hooi Ching; Evander, Mikael; Lilja, Hans; Laurell, Thomas; Scheding, Stefan; Ceder, Yvonne

2018-06-11

Extracellular vesicles (EVs) have emerged as a rich source of biomarkers providing diagnostic and prognostic information in diseases such as cancer. Large-scale investigations into the contents of EVs in clinical cohorts are warranted, but a major obstacle is the lack of a rapid, reproducible, efficient, and low-cost methodology to enrich EVs. Here, we demonstrate the applicability of an automated acoustic-based technique to enrich EVs, termed acoustic trapping. Using this technology, we have successfully enriched EVs from cell culture conditioned media and urine and blood plasma from healthy volunteers. The acoustically trapped samples contained EVs ranging from exosomes to microvesicles in size and contained detectable levels of intravesicular microRNAs. Importantly, this method showed high reproducibility and yielded sufficient quantities of vesicles for downstream analysis. The enrichment could be obtained from a sample volume of 300 μL or less, an equivalent to 30 min of enrichment time, depending on the sensitivity of downstream analysis. Taken together, acoustic trapping provides a rapid, automated, low-volume compatible, and robust method to enrich EVs from biofluids. Thus, it may serve as a novel tool for EV enrichment from large number of samples in a clinical setting with minimum sample preparation.

Determination of Carbonyl Compounds in Cork Agglomerates by GDME-HPLC-UV: Identification of the Extracted Compounds by HPLC-MS/MS.

PubMed

Brandão, Pedro Francisco; Ramos, Rui Miguel; Almeida, Paulo Joaquim; Rodrigues, José António

2017-02-08

A new approach is proposed for the extraction and determination of carbonyl compounds in solid samples, such as wood or cork materials. Cork products are used as building materials due to their singular characteristics; however, little is known about its aldehyde emission potential and content. Sample preparation was done by using a gas-diffusion microextraction (GDME) device for the direct extraction of volatile aldehydes and derivatization with 2,4-dinitrophenylhydrazine. Analytical determination of the extracts was done by HPLC-UV, with detection at 360 nm. The developed methodology proved to be a reliable tool for aldehyde determination in cork agglomerate samples with suitable method features. Mass spectrometry studies were performed for each sample, which enabled the identification, in the extracts, of the derivatization products of a total of 13 aldehydes (formaldehyde, acetaldehyde, furfural, propanal, 5-methylfurfural, butanal, benzaldehyde, pentanal, hexanal, trans-2-heptenal, heptanal, octanal, and trans-2-nonenal) and 4 ketones (3-hydroxy-2-butanone, acetone, cyclohexanone, and acetophenone). This new analytical methodology simultaneously proved to be consistent for the identification and determination of aldehydes in cork agglomerates and a very simple and straightforward procedure.

Studies of the micromorphology of sputtered TiN thin films by autocorrelation techniques

NASA Astrophysics Data System (ADS)

Smagoń, Kamil; Stach, Sebastian; Ţălu, Ştefan; Arman, Ali; Achour, Amine; Luna, Carlos; Ghobadi, Nader; Mardani, Mohsen; Hafezi, Fatemeh; Ahmadpourian, Azin; Ganji, Mohsen; Grayeli Korpi, Alireza

2017-12-01

Autocorrelation techniques are crucial tools for the study of the micromorphology of surfaces: They provide the description of anisotropic properties and the identification of repeated patterns on the surface, facilitating the comparison of samples. In the present investigation, some fundamental concepts of these techniques including the autocorrelation function and autocorrelation length have been reviewed and applied in the study of titanium nitride thin films by atomic force microscopy (AFM). The studied samples were grown on glass substrates by reactive magnetron sputtering at different substrate temperatures (from 25 {}°C to 400 {}°C , and their micromorphology was studied by AFM. The obtained AFM data were analyzed using MountainsMap Premium software obtaining the correlation function, the structure of isotropy and the spatial parameters according to ISO 25178 and EUR 15178N. These studies indicated that the substrate temperature during the deposition process is an important parameter to modify the micromorphology of sputtered TiN thin films and to find optimized surface properties. For instance, the autocorrelation length exhibited a maximum value for the sample prepared at a substrate temperature of 300 {}°C , and the sample obtained at 400 {}°C presented a maximum angle of the direction of the surface structure.

X-ray absorption spectroscopy study of annealing process on Sr1-xLaxCuO2 electron-doped cuprate thin films

NASA Astrophysics Data System (ADS)

Galdi, A.; Orgiani, P.; Sacco, C.; Gobaut, B.; Torelli, P.; Aruta, C.; Brookes, N. B.; Minola, M.; Harter, J. W.; Shen, K. M.; Schlom, D. G.; Maritato, L.

2018-03-01

The superconducting properties of Sr1-xLaxCuO2 thin films are strongly affected by sample preparation procedures, including the annealing step, which are not always well controlled. We have studied the evolution of Cu L2,3 and O K edge x-ray absorption spectra (XAS) of Sr1-xLaxCuO2 thin films as a function of reducing annealing, both qualitatively and quantitatively. By using linearly polarized radiation, we are able to identify the signatures of the presence of apical oxygen in the as-grown sample and its gradual removal as a function of duration of 350 °C Ar annealing performed on the same sample. Even though the as-grown sample appears to be hole doped, we cannot identify the signature of the Zhang-Rice singlet in the O K XAS, and it is extremely unlikely that the interstitial excess oxygen can give rise to a superconducting or even a metallic ground state. XAS and x-ray linear dichroism analyses are, therefore, shown to be valuable tools to improving the control over the annealing process of electron doped superconductors.

Application of semipermeable membrane devices for long-term monitoring of polycyclic aromatic hydrocarbons at various stages of drinking water treatment.

PubMed

Pogorzelec, Marta; Piekarska, Katarzyna

2018-08-01

The primary goal of the presented study was the investigation of occurrence and concentration of sixteen selected polycyclic aromatic hydrocarbons in samples from various stages of water treatment and verification of the applicability of semi-permeable membrane devices in the monitoring of drinking water. Another objective was to verify if weather seasons affect the concentration and complexity of PAHs. For these purposes, semipermeable membrane devices were installed in a surface water treatment plant located in Lower Silesia (Poland). Samples were collected monthly over a period of one year. To determine the effect of water treatment on PAH concentrations, four sampling sites were selected: raw water input, a stream of water in the pipe just before ozonation, treated water output and water after passing through the distribution system. After each month of sampling, SPMDs were exchanged for fresh ones and prepared for instrumental analysis. Concentrations of polycyclic aromatic hydrocarbons were determined by high-performance liquid chromatography (HPLC). The presented study indicates that semipermeable membrane devices can be an effective tool for the analysis of drinking water, in which organic micropollutants occur at very low concentrations. Copyright © 2018 Elsevier B.V. All rights reserved.

A compact LIBS system for industrial applications

NASA Astrophysics Data System (ADS)

Noharet, B.; Sterner, C.; Irebo, T.; Gurell, J.; Bengtson, A.; Vainik, R.; Karlsson, H.; Illy, E.

2015-03-01

In recent years, laser-induced breakdown spectroscopy (LIBS) has been established as a promising analytical tool for online chemical analysis. The emitted light spectrum is analyzed for instantaneous determination of the elemental composition of the sample, enabling on-line classification of materials. Two major strengths of the technique are the possibilities to perform both fast and remote chemical analysis to determine the elemental composition of the samples under test. In order to reduce the size of LIBS systems, the use of a compact Q-switched diode-pumped solid-state laser (DPSSL) in a LIBS system is evaluated for the industrial sorting of aluminium alloys. The DPSSL, which delivers 150μJ pulses of high beam quality at more than 7KHz repetition rate, provides irradiance on the target that is appropriate for LIBS measurements. The experimental results indicate that alloy classification and quantitative analysis are possible on scrap aluminium samples placed 50 cm apart from the focusing and collecting lenses, without sample preparation. Similar calibration curves and limits of detection are obtained for traditional high-energy low-frequency flashlamp-pumped and low-energy high-frequency diode-pumped lasers, showing the applicability of compact diode-pumped lasers for industrial LIBS applications.

Protocols for the analytical characterization of therapeutic monoclonal antibodies. II - Enzymatic and chemical sample preparation.

PubMed

Bobaly, Balazs; D'Atri, Valentina; Goyon, Alexandre; Colas, Olivier; Beck, Alain; Fekete, Szabolcs; Guillarme, Davy

2017-08-15

The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

Sol-gel applications for ceramic membrane preparation

NASA Astrophysics Data System (ADS)

Erdem, I.

2017-02-01

Ceramic membranes possessing superior properties compared to polymeric membranes are more durable under severe working conditions and therefore their service life is longer. The ceramic membranes are composed of some layers. The support is the layer composed of coarser ceramic structure and responsible for mechanical durability under filtration pressure and it is prepared by consolidation of ceramic powders. The top layer is composed of a finer ceramic micro-structure mainly responsible for the separation of components present in the fluid to be filtered and sol-gel method is a versatile tool to prepare such a tailor-made ceramic filtration structure with finer pores. Depending on the type of filtration (e.g. micro-filtration, ultra-filtration, nano-filtration) aiming separation of components with different sizes, sols with different particulate sizes should be prepared and consolidated with varying precursors and preparation conditions. The coating of sol on the support layer and heat treatment application to have a stable ceramic micro-structure are also important steps determining the final properties of the top layer. Sol-gel method with various controllable parameters (e.g. precursor type, sol formation kinetics, heat treatment conditions) is a practical tool for the preparation of top layers of ceramic composite membranes with desired physicochemical properties.

Microstructural and mechanical characteristics of porous iron prepared by powder metallurgy.

PubMed

Capek, Jaroslav; Vojtěch, Dalibor

2014-10-01

The demand for porous biodegradable load-bearing implants has been increasing recently. Based on investigations of biodegradable stents, porous iron may be a suitable material for such applications. In this study, we prepared porous iron samples with porosities of 34-51 vol.% by powder metallurgy using ammonium bicarbonate as a space-holder material. We studied sample microstructure (SEM-EDX and XRD), flexural and compressive behaviors (universal loading machine) and hardness HV5 (hardness tester) of the prepared samples. Sample porosity increased with the amount of spacer in the initial mixtures. Only the pore surfaces had insignificant oxidation and no other contamination was observed. Increasing porosity decreased the mechanical properties of the samples; although, the properties were still comparable with human bone and higher than those of porous non-metallic biomaterials and porous magnesium prepared in a similar way. Based on these results, powder metallurgy appears to be a suitable method for the preparation of porous iron for orthopedic applications. Copyright © 2014 Elsevier B.V. All rights reserved.

Synthesis of SrFe12O19 magnetic nanoparticles by EDTA complex method

NASA Astrophysics Data System (ADS)

Wang, Shifa; Li, Danming; Xiao, Yuhua; Dang, Wenqiang; Feng, Jie

2017-10-01

A modified polyacrylamide gel route was used to prepare SrFe12O19 magnetic nanoparticles; ethylenediaminetetraacetic acid (EDTA) was used as a carboxyl chelating agent. The phase purity, morphology and magnetic properties of as-prepared samples were analyzed via X-ray diffraction (XRD), Raman spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibrating sample magnetometery (VSM). XRD analysis indicates that high-purity SrFe12O19 magnetic nanoparticles can be synthesized at 700°C in air. The characteristic peaks of as-prepared sample at 210, 283, 321, 340, 381, 411, 432, 475, 532, 618, 686, and 726 cm-1 were observed in Raman spectra. SEM and TEM show that the synthesized SrFe12O19 magnetic nanoparticles are uniform with the mean particle size of 60 nm. VSM measurement shows that the maximum magnetic energy product (BH)max of sample prepared using EDTA as a chelating agent is higher than that of sample prepared using citric acid as a chelating agent.

'FloraArray' for screening of specific DNA probes representing the characteristics of a certain microbial community.

PubMed

Yokoi, Takahide; Kaku, Yoshiko; Suzuki, Hiroyuki; Ohta, Masayuki; Ikuta, Hajime; Isaka, Kazuichi; Sumino, Tatsuo; Wagatsuma, Masako

2007-08-01

To investigate uncharacterized microbial communities, a custom DNA microarray named 'FloraArray' was developed for screening specific probes that would represent the characteristics of a microbial community. The array was prepared by spotting 2000 plasmid DNAs from a genomic shotgun library of a sludge sample on a DNA microarray. By comparative hybridization of the array with two different samples of genomic DNA, one from the activated sludge and the other from a nonactivated sludge sample of an anaerobic ammonium oxidation (anammox) bacterial community, specific spots were visualized as a definite fluctuating profile in an MA (differential intensity ratio vs. spot intensity) plot. About 300 spots of the array accounted for the candidate probes to represent anammox reaction of the activated sludge. After sequence analysis of the probes and examination of the results of blastn searches against the reported anammox reference sequence, complete matches were found for 161 probes (58.3%) and >90% matches were found for 242 probes (87.1%). These results demonstrate that 'FloraArray' could be a useful tool for screening specific DNA molecules of unknown microbial communities.

The OSIRIS-REx Mission Sample Site Selection Process

NASA Astrophysics Data System (ADS)

Beshore, Edward C.; Lauretta, Dante

2014-11-01

In September of 2016, the OSIRIS-REx (Origins, Spectral Interpretation, Resource Identification, Security, REgolith eXplorer) spacecraft will depart for asteroid (101955) Bennu, and in doing so, will turn an important corner in the exploration of the solar system. After arriving at Bennu in the fall of 2018, OSIRIS-REx will undertake a program of observations designed to select a site suitable for retrieving a sample that will be returned to the Earth in 2023. The third mission in NASA’s New Frontiers program, OSIRIS-REx will return over 60 grams from Bennu’s surface.OSIRIS-REx is unique because the science team will have an operational role to play in preparing data products needed to select a sample site. These include products used to ensure flight system safety — topographic maps and shape models, temperature measurements, maps of hazards — as well as assessments of sampleability and science value. The timing and production of these will be presented, as will the high-level decision-making tools and processes for the interim and final site selection processes.

Increasing productivity for the analysis of trace contaminants in food by gas chromatography-mass spectrometry using automated liner exchange, backflushing and heart-cutting.

PubMed

David, Frank; Tienpont, Bart; Devos, Christophe; Lerch, Oliver; Sandra, Pat

2013-10-25

Laboratories focusing on residue analysis in food are continuously seeking to increase sample throughput by minimizing sample preparation. Generic sample extraction methods such as QuEChERS lack selectivity and consequently extracts are not free from non-volatile material that contaminates the analytical system. Co-extracted matrix constituents interfere with target analytes, even if highly sensitive and selective GC-MS/MS is used. A number of GC approaches are described that can be used to increase laboratory productivity. These techniques include automated inlet liner exchange and column backflushing for preservation of the performance of the analytical system and heart-cutting two-dimensional GC for increasing sensitivity and selectivity. The application of these tools is illustrated by the analysis of pesticides in vegetables and fruits, PCBs in milk powder and coplanar PCBs in fish. It is demonstrated that considerable increase in productivity can be achieved by decreasing instrument down-time, while analytical performance is equal or better compared to conventional trace contaminant analysis. Copyright © 2013 Elsevier B.V. All rights reserved.

Identification of Microbial Profile of Koji Using Single Molecule, Real-Time Sequencing Technology.

PubMed

Hui, Wenyan; Hou, Qiangchuan; Cao, Chenxia; Xu, Haiyan; Zhen, Yi; Kwok, Lai-Yu; Sun, Tiansong; Zhang, Heping; Zhang, Wenyi

2017-05-01

Koji is a kind of Japanese traditional fermented starter that has been used for centuries. Many fermented foods are made from koji, such as sake, miso, and soy sauce. This study used the single molecule real-time sequencing technology (SMRT) to investigate the bacterial and fungal microbiota of 3 Japanese koji samples. After SMRT analysis, a total of 39121 high-quality sequences were generated, including 14354 bacterial and 24767 fungal sequence reads. The high-quality gene sequences were assigned to 5 bacterial and 2 fungal plyla, dominated by Proteobacteria and Ascomycota, respectively. At the genus level, Ochrobactrum and Wickerhamomyces were the most abundant bacterial and fungal genera, respectively. The predominant bacterial and fungal species were Ochrobactrum lupini and Wickerhamomyces anomalus, respectively. Our study profiled the microbiota composition of 3 Japanese koji samples to the species level precision. The results may be useful for further development of traditional fermented products, especially optimization of koji preparation. Meanwhile, this study has demonstrated that SMRT is a robust tool for analyzing the microbial composition in food samples. © 2017 Institute of Food Technologists®.

Investigation of cloud point extraction for the analysis of metallic nanoparticles in a soil matrix

PubMed Central

Hadri, Hind El; Hackley, Vincent A.

2017-01-01

The characterization of manufactured nanoparticles (MNPs) in environmental samples is necessary to assess their behavior, fate and potential toxicity. Several techniques are available, but the limit of detection (LOD) is often too high for environmentally relevant concentrations. Therefore, pre-concentration of MNPs is an important component in the sample preparation step, in order to apply analytical tools with a LOD higher than the ng kg−1 level. The objective of this study was to explore cloud point extraction (CPE) as a viable method to pre-concentrate gold nanoparticles (AuNPs), as a model MNP, spiked into a soil extract matrix. To that end, different extraction conditions and surface coatings were evaluated in a simple matrix. The CPE method was then applied to soil extract samples spiked with AuNPs. Total gold, determined by inductively coupled plasma mass spectrometry (ICP-MS) following acid digestion, yielded a recovery greater than 90 %. The first known application of single particle ICP-MS and asymmetric flow field-flow fractionation to evaluate the preservation of the AuNP physical state following CPE extraction is demonstrated. PMID:28507763

LC-MS based analysis of endogenous steroid hormones in human hair.

PubMed

Gao, Wei; Kirschbaum, Clemens; Grass, Juliane; Stalder, Tobias

2016-09-01

The quantification of endogenous steroid hormone concentrations in hair is increasingly used as a method for obtaining retrospective information on long-term integrated hormone exposure. Several different analytical procedures have been employed for hair steroid analysis, with liquid chromatography-mass spectrometry (LC-MS) being recognized as a particularly powerful analytical tool. Several methodological aspects affect the performance of LC-MS systems for hair steroid analysis, including sample preparation and pretreatment, steroid extraction, post-incubation purification, LC methodology, ionization techniques and MS specifications. Here, we critically review the differential value of such protocol variants for hair steroid hormones analysis, focusing on both analytical quality and practical feasibility issues. Our results show that, when methodological challenges are adequately addressed, LC-MS protocols can not only yield excellent sensitivity and specificity but are also characterized by relatively simple sample processing and short run times. This makes LC-MS based hair steroid protocols particularly suitable as a high-quality option for routine application in research contexts requiring the processing of larger numbers of samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

Predictive engineering tools for injection-molded long-carbon-fiber thermoplastic composites - FY 2015 third quarterly report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nguyen, Ba Nghiep; Fifield, Leonard S.; Mori, Steven

During the third quarter of FY 2015, the following technical progress has been made toward project milestones: 1) Magna oversaw the tool build and prepared the molding plan for the complex part of Phase II. 2) PlastiComp hosted a visit by Magna and Toyota on April 23rd to finalize the molding scope and schedule. The plan for molding trials including selection of molding parameters for both LFT and D-LFT for the U-shape complex part was established. 3) Toyota shipped the U-shape complex part tool to Magna on May 28th, 2015. 4) Plasticomp provided 30wt% LCF/PP and 30wt% LCF/PA66 compounded pelletsmore » to Magna for molding the complex part. 5) Magna performed preliminary molding trials on June 2nd, 2015 to validate wall thickness, fill profile, tool temperature and shot size requirements for the complex part. 6) Magna performed the first complex part run on June 16th and 17th, 2015 at Magna’s Composite Centre of Excellence in Concord, ON, Canada. Dale Roland of Plasticomp, and Umesh Gandhi of Toyota also attended the molding. 7) Magna discussed and finalized the plan with PNNL and the team for cutting samples from molded parts at selected locations for fiber orientation and length measurements. 8) Magna provided the computer-aided design (CAD) files of the complex parts with and without ribs to PNNL and Autodesk to build the corresponding ASMI models for injection molding simulations. Magna also provided the actual parameters used. 9) Plasticomp’s provided knowledge and experience of molding LCF materials essential to the successful molding of the parts including optimization of fill speed, tool temperatures, and plasticizing conditions for the 30wt% LCF/PP and 30wt% LCF/PA66 materials in both rib and non-rib versions. 10) Magna molded additional parts for evaluation of mechanical property testing including torsional stiffness on June 29th and 30th, 2015 at Magna’s Composite Center of Excellence. 11) Toyota began preparation for the torsion test of the specimens. Preparation of a computer-aided engineering (CAE) model to predict the performance is in progress. 12) Autodesk fixed an error in the implementation of the proper orthogonal decomposition (POD) calculation of fiber length that had caused the ASMI solution to crash and provided an updated build of ASMI containing the fix. 13)Autodesk reviewed and provided feedback for the complex part molding and measurement locations. 14) Autodesk provided support to set up the workflow for ASMI-ABAQUS® analysis, and provided a fix and workaround for a bug in the ASMI-ABAQUS® output command. 15) Autodesk helped build ASMI analysis models for the complex parts with and without ribs. 16) Autodesk worked on improving the orientation prediction accuracy in the shearing layer for 3D meshes based on comparison to measured data of the plaque moldings. 17) PNNL installed a new ASMI version received from Autodesk and performed comparative analyses to assess mid-plane versus 3D fiber length predictions using the full fiber length model and the reduced-order model (ROM) using POD. 18) PNNL presented the project scope, accomplishments, significant results and future plans to DOE and the USCAR Materials Tech Team on June 3rd, 2015. 19) PNNL discussed the cutting of samples from molded parts and finalized a plan with Magna and the team suggesting the sample size, locations and number of samples per location. 20) PNNL and Autodesk built ASMI models for the complex parts with and without ribs, and preliminary analyses of the part with ribs were conducted using the actual molding parameters received from Magna. 21) PNNL worked on a procedure to extract fiber orientation and length results from a 3D ASMI analysis to a 3D ABAQUS model. This procedure is essential to import ASMI fiber orientation and length to a 3D ABAQUS model of the part allowing future part structural analysis for weight reduction study.« less

Chip-Based Sensors for Disease Diagnosis

NASA Astrophysics Data System (ADS)

Fang, Zhichao

Nucleic acid analysis is one of the most important disease diagnostic approaches in medical practice, and has been commonly used in cancer biomarker detection, bacterial speciation and many other fields in laboratory. Currently, the application of powerful research methods for genetic analysis, including the polymerase chain reaction (PCR), DNA sequencing, and gene expression profiling using fluorescence microarrays, are not widely used in hospitals and extended-care units due to high-cost, long detection times, and extensive sample preparation. Bioassays, especially chip-based electrochemical sensors, may be suitable for the next generation of rapid, sensitive, and multiplexed detection tools. Herein, we report three different microelectrode platforms with capabilities enabled by nano- and microtechnology: nanoelectrode ensembles (NEEs), nanostructured microelectrodes (NMEs), and hierarchical nanostructured microelectrodes (HNMEs), all of which are able to directly detect unpurified RNA in clinical samples without enzymatic amplification. Biomarkers that are cancer and infectious disease relevant to clinical medicine were chosen to be the targets. Markers were successfully detected with clinically-relevant sensitivity. Using peptide nucleic acids (PNAs) as probes and an electrocatalytic reporter system, NEEs were able to detect prostate cancer-related gene fusions in tumor tissue samples with 100 ng of RNA. The development of NMEs improved the sensitivity of the assay further to 10 aM of DNA target, and multiplexed detection of RNA sequences of different prostate cancer-related gene fusion types was achieved on the chip-based NMEs platform. An HNMEs chip integrated with a bacterial lysis device was able to detect as few as 25 cfu bacteria in 30 minutes and monitor the detection in real time. Bacterial detection could also be performed in neat urine samples. The development of these versatile clinical diagnostic tools could be extended to the detection of various cancers, genetic, and infectious diseases.

Discrimination of gastrointestinal nematode eggs from crude fecal egg preparations by inhibitor-resistant conventional and real-time PCR.

PubMed

Demeler, Janina; Ramünke, Sabrina; Wolken, Sonja; Ianiello, Davide; Rinaldi, Laura; Gahutu, Jean Bosco; Cringoli, Giuseppe; von Samson-Himmelstjerna, Georg; Krücken, Jürgen

2013-01-01

Diagnosis of gastrointestinal nematodes relies predominantly on coproscopic methods such as flotation, Kato-Katz, McMaster or FLOTAC. Although FLOTAC allows accurate quantification, many nematode eggs can only be differentiated to genus or family level. Several molecular diagnostic tools discriminating closely related species suffer from high costs for DNA isolation from feces and limited sensitivity since most kits use only small amounts of feces (<1 g). A direct PCR from crude egg preparations was designed for full compatibility with FLOTAC to accurately quantify eggs per gram feces (epg) and determine species composition. Eggs were recovered from the flotation solution and concentrated by sieving. Lysis was achieved by repeated boiling and freezing cycles - only Trichuris eggs required additional mechanic disruption. Egg lysates were directly used as template for PCR with Phusion DNA polymerase which is particularly resistant to PCR inhibitors. Qualitative results were obtained with feces of goats, cattle, horses, swine, cats, dogs and mice. The finally established protocol was also compatible with quantitative real-time PCR in the presence of EvaGreen and no PCR inhibition was detectable when extracts were diluted at least fourfold. Sensitivity was comparable to DNA isolation protocols and spiked samples with five epg were reliably detected. For Toxocara cati a detection limit below one epg was demonstrated. It was possible to distinguish T. cati and Toxocara canis using high resolution melt (HRM) analysis, a rapid tool for species identification. In human samples, restriction fragment length polymorphism (RFLP) and HRM analysis were used to discriminate Necator americanus and Ancylostoma duodenale. The method is able to significantly improve molecular diagnosis of gastrointestinal nematodes by increasing speed and sensitivity while decreasing overall costs. For identification of species or resistance alleles, analysis of PCR products with many different post PCR methods can be used such as RFLP, reverse-line-blot, Sanger sequencing and HRM.

Monoolein Lipid Phases as Incorporation and Enrichment Materials for Membrane Protein Crystallization

PubMed Central

Wallace, Ellen; Dranow, David; Laible, Philip D.; Christensen, Jeff; Nollert, Peter

2011-01-01

The crystallization of membrane proteins in amphiphile-rich materials such as lipidic cubic phases is an established methodology in many structural biology laboratories. The standard procedure employed with this methodology requires the generation of a highly viscous lipidic material by mixing lipid, for instance monoolein, with a solution of the detergent solubilized membrane protein. This preparation is often carried out with specialized mixing tools that allow handling of the highly viscous materials while minimizing dead volume to save precious membrane protein sample. The processes that occur during the initial mixing of the lipid with the membrane protein are not well understood. Here we show that the formation of the lipidic phases and the incorporation of the membrane protein into such materials can be separated experimentally. Specifically, we have investigated the effect of different initial monoolein-based lipid phase states on the crystallization behavior of the colored photosynthetic reaction center from Rhodobacter sphaeroides. We find that the detergent solubilized photosynthetic reaction center spontaneously inserts into and concentrates in the lipid matrix without any mixing, and that the initial lipid material phase state is irrelevant for productive crystallization. A substantial in-situ enrichment of the membrane protein to concentration levels that are otherwise unobtainable occurs in a thin layer on the surface of the lipidic material. These results have important practical applications and hence we suggest a simplified protocol for membrane protein crystallization within amphiphile rich materials, eliminating any specialized mixing tools to prepare crystallization experiments within lipidic cubic phases. Furthermore, by virtue of sampling a membrane protein concentration gradient within a single crystallization experiment, this crystallization technique is more robust and increases the efficiency of identifying productive crystallization parameters. Finally, we provide a model that explains the incorporation of the membrane protein from solution into the lipid phase via a portal lamellar phase. PMID:21909395

A method for screening active components from Chinese herbs by cell membrane chromatography-offline-high performance liquid chromatography/mass spectrometry and an online statistical tool for data processing.

PubMed

Cao, Yan; Wang, Shaozhan; Li, Yinghua; Chen, Xiaofei; Chen, Langdong; Wang, Dongyao; Zhu, Zhenyu; Yuan, Yongfang; Lv, Diya

2018-03-09

Cell membrane chromatography (CMC) has been successfully applied to screen bioactive compounds from Chinese herbs for many years, and some offline and online two-dimensional (2D) CMC-high performance liquid chromatography (HPLC) hyphenated systems have been established to perform screening assays. However, the requirement of sample preparation steps for the second-dimensional analysis in offline systems and the need for an interface device and technical expertise in the online system limit their extensive use. In the present study, an offline 2D CMC-HPLC analysis combined with the XCMS (various forms of chromatography coupled to mass spectrometry) Online statistical tool for data processing was established. First, our previously reported online 2D screening system was used to analyze three Chinese herbs that were reported to have potential anti-inflammatory effects, and two binding components were identified. By contrast, the proposed offline 2D screening method with XCMS Online analysis was applied, and three more ingredients were discovered in addition to the two compounds revealed by the online system. Then, cross-validation of the three compounds was performed, and they were confirmed to be included in the online data as well, but were not identified there because of their low concentrations and lack of credible statistical approaches. Last, pharmacological experiments showed that these five ingredients could inhibit IL-6 release and IL-6 gene expression on LPS-induced RAW cells in a dose-dependent manner. Compared with previous 2D CMC screening systems, this newly developed offline 2D method needs no sample preparation steps for the second-dimensional analysis, and it is sensitive, efficient, and convenient. It will be applicable in identifying active components from Chinese herbs and practical in discovery of lead compounds derived from herbs. Copyright © 2018 Elsevier B.V. All rights reserved.

Discrimination of Gastrointestinal Nematode Eggs from Crude Fecal Egg Preparations by Inhibitor-Resistant Conventional and Real-Time PCR

PubMed Central

Demeler, Janina; Ramünke, Sabrina; Wolken, Sonja; Ianiello, Davide; Rinaldi, Laura; Gahutu, Jean Bosco; Cringoli, Giuseppe; von Samson-Himmelstjerna, Georg; Krücken, Jürgen

2013-01-01

Diagnosis of gastrointestinal nematodes relies predominantly on coproscopic methods such as flotation, Kato-Katz, McMaster or FLOTAC. Although FLOTAC allows accurate quantification, many nematode eggs can only be differentiated to genus or family level. Several molecular diagnostic tools discriminating closely related species suffer from high costs for DNA isolation from feces and limited sensitivity since most kits use only small amounts of feces (<1 g). A direct PCR from crude egg preparations was designed for full compatibility with FLOTAC to accurately quantify eggs per gram feces (epg) and determine species composition. Eggs were recovered from the flotation solution and concentrated by sieving. Lysis was achieved by repeated boiling and freezing cycles – only Trichuris eggs required additional mechanic disruption. Egg lysates were directly used as template for PCR with Phusion DNA polymerase which is particularly resistant to PCR inhibitors. Qualitative results were obtained with feces of goats, cattle, horses, swine, cats, dogs and mice. The finally established protocol was also compatible with quantitative real-time PCR in the presence of EvaGreen and no PCR inhibition was detectable when extracts were diluted at least fourfold. Sensitivity was comparable to DNA isolation protocols and spiked samples with five epg were reliably detected. For Toxocara cati a detection limit below one epg was demonstrated. It was possible to distinguish T. cati and Toxocara canis using high resolution melt (HRM) analysis, a rapid tool for species identification. In human samples, restriction fragment length polymorphism (RFLP) and HRM analysis were used to discriminate Necator americanus and Ancylostoma duodenale. The method is able to significantly improve molecular diagnosis of gastrointestinal nematodes by increasing speed and sensitivity while decreasing overall costs. For identification of species or resistance alleles, analysis of PCR products with many different post PCR methods can be used such as RFLP, reverse-line-blot, Sanger sequencing and HRM. PMID:23620739

School Turnaround Teachers: Selection Toolkit. Part of the School Turnaround Collection from Public Impact

ERIC Educational Resources Information Center

Public Impact, 2008

2008-01-01

This toolkit includes these separate sections: (1) Selection Preparation Guide; (2) Day-of-Interview Tools; (3) Candidate Rating Tools; and (4) Candidate Comparison and Decision Tools. Each of the sections is designed to be used at different stages of the selection process. The first section provides turnaround teacher competencies that are the…

School Turnaround Leaders: Selection Toolkit. Part of the School Turnaround Collection from Public Impact

ERIC Educational Resources Information Center

Public Impact, 2008

2008-01-01

This toolkit includes the following separate sections: (1) Selection Preparation Guide; (2) Day-of-Interview Tools; (3) Candidate Rating Tools; and (4) Candidate Comparison and Decision Tools. Each of the sections is designed to be used at different stages of the selection process. The first section provides a list of competencies that would…

Standardizing the experimental conditions for using urine in NMR-based metabolomic studies with a particular focus on diagnostic studies: a review.

PubMed

Emwas, Abdul-Hamid; Luchinat, Claudio; Turano, Paola; Tenori, Leonardo; Roy, Raja; Salek, Reza M; Ryan, Danielle; Merzaban, Jasmeen S; Kaddurah-Daouk, Rima; Zeri, Ana Carolina; Nagana Gowda, G A; Raftery, Daniel; Wang, Yulan; Brennan, Lorraine; Wishart, David S

The metabolic composition of human biofluids can provide important diagnostic and prognostic information. Among the biofluids most commonly analyzed in metabolomic studies, urine appears to be particularly useful. It is abundant, readily available, easily stored and can be collected by simple, noninvasive techniques. Moreover, given its chemical complexity, urine is particularly rich in potential disease biomarkers. This makes it an ideal biofluid for detecting or monitoring disease processes. Among the metabolomic tools available for urine analysis, NMR spectroscopy has proven to be particularly well-suited, because the technique is highly reproducible and requires minimal sample handling. As it permits the identification and quantification of a wide range of compounds, independent of their chemical properties, NMR spectroscopy has been frequently used to detect or discover disease fingerprints and biomarkers in urine. Although protocols for NMR data acquisition and processing have been standardized, no consensus on protocols for urine sample selection, collection, storage and preparation in NMR-based metabolomic studies have been developed. This lack of consensus may be leading to spurious biomarkers being reported and may account for a general lack of reproducibility between laboratories. Here, we review a large number of published studies on NMR-based urine metabolic profiling with the aim of identifying key variables that may affect the results of metabolomics studies. From this survey, we identify a number of issues that require either standardization or careful accounting in experimental design and provide some recommendations for urine collection, sample preparation and data acquisition.

Evaluation of a recent product to remove lipids and other matrix co-extractives in the analysis of pesticide residues and environmental contaminants in foods.

PubMed

Han, Lijun; Matarrita, Jessie; Sapozhnikova, Yelena; Lehotay, Steven J

2016-06-03

This study demonstrates the application of a novel lipid removal product to the residue analysis of 65 pesticides and 52 environmental contaminants in kale, pork, salmon, and avocado by fast, low pressure gas chromatography - tandem mass spectrometry (LPGC-MS/MS). Sample preparation involves QuEChERS extraction followed by use of EMR-Lipid ("enhanced matrix removal of lipids") and an additional salting out step for cleanup. The optimal amount of EMR-Lipid was determined to be 500mg for 2.5mL extracts for most of the analytes. The co-extractive removal efficiency by the EMR-Lipid cleanup step was 83-98% for fatty samples and 79% for kale, including 76% removal of chlorophyll. Matrix effects were typically less than ±20%, in part because analyte protectants were used in the LPGC-MS/MS analysis. The recoveries of polycyclic aromatic hydrocarbons and diverse pesticides were mostly 70-120%, whereas recoveries of nonpolar polybrominated diphenyl ethers and polychlorinated biphenyls were mostly lower than 70% through the cleanup procedure. With the use of internal standards, method validation results showed that 76-85 of the 117 analytes achieved satisfactory results (recoveries of 70-120% and RSD≤20%) in pork, avocado, and kale, while 53 analytes had satisfactory results in salmon. Detection limits were 5-10ng/g for all but a few analytes. EMR-Lipid is a new sample preparation tool that serves as another useful option for cleanup in multiresidue analysis, particularly of fatty foods. Published by Elsevier B.V.

Rapid screening of abused drugs by direct analysis in real time (DART) coupled to time-of-flight mass spectrometry (TOF-MS) combined with ion mobility spectrometry (IMS).

PubMed

Lian, Ru; Wu, Zhongping; Lv, Xiaobao; Rao, Yulan; Li, Haiyang; Li, Jinghua; Wang, Rong; Ni, Chunfang; Zhang, Yurong

2017-10-01

Increasing in cases involving drugs of abuse leads to heavy burden for law enforcement agencies, exacerbating demand for rapid screening technique. In this study, atmospheric pressure ionization technologies including direct analysis in real time (DART) ion source coupled to a time-of-flight mass spectrometer (DART-TOF-MS)as well asdopant-assisted positive photoionization ion mobility spectrometry (DAPP-IMS) without radioactivity were utilized together as the powerful analytical tool for the rapid screening and identification of 53 abused drugs.The limits of detection (LOD) were 0.05-2μg/mL when using DART-TOF-MS and 0.02-2μg when using DAPP-IMS which could satisfy the actual requirement in forensic science laboratory. The advantages of this method included fast response, high-throughput potential, high specificity, and minimal sample preparation. A screening library of reduced mobility (K 0 ), accurate mass of informative precursor ion ([M+H] + ) and fragment ions was established respectively by employing a bench-top DAPP-IMS and TOF-MS in-source collision induced dissociation (CID) mode. Then the standardized screening procedure was developed with criteria for the confirmation of positive result. A total of 50 seized drug samples provided by local forensic laboratory we reanalyzed to testify the utility of the method. This study suggests that a method combing DART-TOF-MS and DAPP-IMS is promising for the rapid screening and identification of abused drugs with minimal sample preparation and absence of chromatography. Copyright © 2017 Elsevier B.V. All rights reserved.

Analysis and differentiation of paper samples by capillary electrophoresis and multivariate analysis.

PubMed

Fernández de la Ossa, Ma Ángeles; Ortega-Ojeda, Fernando; García-Ruiz, Carmen

2014-11-01

This work reports an investigation for the analysis of different paper samples using CE with laser-induced detection. Papers from four different manufactures (white-copy paper) and four different paper sources (white and recycled-copy papers, adhesive yellow paper notes and restaurant serviettes) were pulverized by scratching with a surgical scalpel prior to their derivatization with a fluorescent labeling agent, 8-aminopyrene-1,3,6-trisulfonic acid. Methodological conditions were evaluated, specifically the derivatization conditions with the aim to achieve the best S/N signals and the separation conditions in order to obtain optimum values of sensitivity and reproducibility. The best conditions, in terms of fastest, and easiest sample preparation procedure, minimal sample consumption, as well as the use of the simplest and fastest CE-procedure for obtaining the best analytical parameters, were applied to the analysis of the different paper samples. The registered electropherograms were pretreated (normalized and aligned) and subjected to multivariate analysis (principal component analysis). A successful discrimination among paper samples without entanglements was achieved. To the best of our knowledge, this work presents the first approach to achieve a successful differentiation among visually similar white-copy paper samples produced by different manufactures and paper from different paper sources through their direct analysis by CE-LIF and subsequent comparative study of the complete cellulose electropherogram by chemometric tools. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Monolithic methacrylate packed 96-tips for high throughput bioanalysis.

PubMed

Altun, Zeki; Skoglund, Christina; Abdel-Rehim, Mohamed

2010-04-16

In the pharmaceutical industry the growing number of samples to be analyzed requires high throughput and fully automated analytical techniques. Commonly used sample-preparation methods are solid-phase extraction (SPE), liquid-liquid extraction (LLE) and protein precipitation. In this paper we will discus a new sample-preparation technique based on SPE for high throughput drug extraction developed and used by our group. This new sample-preparation method is based on monolithic methacrylate polymer as packing sorbent for 96-tip robotic device. Using this device a 96-well plate could be handled in 2-4min. The key aspect of the monolithic phase is that monolithic material can offer both good binding capacity and low back-pressure properties compared to e.g. silica phases. The present paper presents the successful application of monolithic 96-tips and LC-MS/MS by the sample preparation of busulphan, rescovitine, metoprolol, pindolol and local anaesthetics from human plasma samples and cyklophosphamid from mice blood samples. Copyright 2009 Elsevier B.V. All rights reserved.

Electrodeposition as an alternate method for preparation of environmental samples for iodide by AMS

DOE PAGES

Adamic, M. L.; Lister, T. E.; Dufek, E. J.; ...

2015-03-25

This paper presents an evaluation of an alternate method for preparing environmental samples for 129I analysis by accelerator mass spectrometry (AMS) at Idaho National Laboratory. The optimal sample preparation method is characterized by ease of preparation, capability of processing very small quantities of iodide, and ease of loading into a cathode. Electrodeposition of iodide on a silver wire was evaluated using these criteria. This study indicates that the electrochemically-formed silver iodide deposits produce ion currents similar to those from precipitated silver iodide for the same sample mass. Furthermore, precipitated silver iodide samples are usually mixed with niobium or silver powdermore » prior to loading in a cathode. Using electrodeposition, the silver is already mixed with the sample and can simply be picked up with tweezers, placed in the sample die, and pressed into a cathode. The major advantage of this method is that the silver wire/electrodeposited silver iodide is much easier to load into a cathode.« less

(Project 13-5292) Correlating thermal and mechanical coupling based multiphysics behavior of nuclear materials through in-situ measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tomar, Vikas

Irradiations and post characterization experiments were performed first on Zr samples. This step will help understand the effect of the 2.5% alloying elements on the behavior of Zircaloy-4 (PWR cladding material) when compared to pure Zr. Irradiation flux measurements and sample temperature calibrations were performed at different energies prior to the irradiation experiments. Irradiations were performed with two different energy regimes1: non-displacment energies and displacement energies. Time was also dedicated to optimize transmission electron microscopy (TEM) sample preparation conditions via electropolishing technique. This step is crucial to prepare TEM samples for the in-situ TEM/irradiation experiments (Year 2). In addition, Zircaloy-4more » samples are being prepared for irradiation, and a setup is built by one of our collaborators (Dr. Mert Efe) to prepare ultrafine (UF) and nanocrystalline (NC) Zircaloy-4 samples for comparison with the commercial Zircaloy-4 samples.« less

Media preparation and bacteriological tools.

PubMed

Elbing, Karen; Brent, Roger

2002-08-01

Recipes are provided in this unit for minimal liquid media, rich liquid media, solid media, top agar, and stab agar. Also included are descriptions and useful information about tools used with growth media such as inoculating loops, sterile toothpicks and spreaders.

A Five-Year Evaluation of Examination Structure in a Cardiovascular Pharmacotherapy Course

PubMed Central

Kolar, Claire; Janke, Kristin K.

2015-01-01

Objective. To evaluate the composition and effectiveness as an assessment tool of a criterion-referenced examination comprised of clinical cases tied to practice decisions, to examine the effect of varying audience response system (ARS) questions on student examination preparation, and to articulate guidelines for structuring examinations to maximize evaluation of student learning. Design. Multiple-choice items developed over 5 years were evaluated using Bloom’s Taxonomy classification, point biserial correlation, item difficulty, and grade distribution. In addition, examination items were classified into categories based on similarity to items used in ARS preparation. Assessment. As the number of items directly tied to clinical practice rose, Bloom’s Taxonomy level and item difficulty also rose. In examination years where Bloom’s levels were high but preparation was minimal, average grade distribution was lower compared with years in which student preparation was higher. Conclusion. Criterion-referenced examinations can benefit from systematic evaluation of their composition and effectiveness as assessment tools. Calculated design and delivery of classroom preparation is an asset in improving examination performance on rigorous, practice-relevant examinations. PMID:27168611

Analysis of nutrition-relevant trace elements in human blood and serum by means of total reflection X-ray fluorescence (TXRF) spectroscopy

NASA Astrophysics Data System (ADS)

Stosnach, Hagen; Mages, Margarete

2009-04-01

In clinical service laboratories, one of the most common analytical tasks with regard to inorganic traces is the determination of the nutrition-relevant elements Fe, Cu, Zn, and Se. Because of the high numbers of samples and the commercial character of these analyses, a time-consuming sample preparation must be avoided. In this presentation, the results of total reflection X-ray fluorescence measurements with a low-power system and different sample preparation procedures are compared with those derived from analysis with common methods like Atomic Absorption Spectroscopy (AAS) and Inductively Coupled Plasma Mass Spectroscopy (ICP-MS). The results of these investigations indicate that the optimal total reflection X-ray fluorescence analysis of the nutrition-relevant elements Fe, Cu, Zn, and Se can be performed by preparing whole blood and serum samples after dilution with ultrapure water and transferring 10 μl of internally standardized sample to an unsiliconized quartz glass sample carrier with subsequent drying in a laboratory oven. Suitable measurement time was found to be 600 s. The enhanced sample preparation by means of microwave or open digestion, in parts combined with cold plasma ashing, led to an improvement of detection limits by a factor of 2 for serum samples while for whole blood samples an improvement was only observed for samples prepared by means of microwave digestion. As the matrix elements P, S, Cl, and for whole blood Fe have a major influence on the detection limits, most probably a further enhancement of analytical quality requires the removal of the organic matrix. However, for the routine analysis of the nutrition-relevant elements, the dilution preparation was found to be sufficient.

Preparation of poly-L-lysine functionalized magnetic nanoparticles and their influence on viability of cancer cells

NASA Astrophysics Data System (ADS)

Khmara, I.; Koneracka, M.; Kubovcikova, M.; Zavisova, V.; Antal, I.; Csach, K.; Kopcansky, P.; Vidlickova, I.; Csaderova, L.; Pastorekova, S.; Zatovicova, M.

2017-04-01

This study was aimed at development of biocompatible amino-functionalized magnetic nanoparticles as carriers of specific antibodies able to detect and/or target cancer cells. Poly-L-lysine (PLL)-modified magnetic nanoparticle samples with different PLL/Fe3O4 content were prepared and tested to define the optimal PLL/Fe3O4 weight ratio. The samples were characterized for particle size and morphology (SEM, TEM and DLS), and surface properties (zeta potential measurements). The optimal PLL/Fe3O4 weight ratio of 1.0 based on both zeta potential and DLS measurements was in agreement with the UV/VIS measurements. Magnetic nanoparticles with the optimal PLL content were conjugated with antibody specific for the cancer biomarker carbonic anhydrase IX (CA IX), which is induced by hypoxia, a physiologic stress present in solid tumors and linked with aggressive tumor behavior. CA IX is localized on the cell surface with the antibody-binding epitope facing the extracellular space and is therefore suitable for antibody-based targeting of tumor cells. Here we showed that PLL/Fe3O4 magnetic nanoparticles exhibit cytotoxic activities in a cell type-dependent manner and bind to cells expressing CA IX when conjugated with the CA IX-specific antibody. These data support further investigations of the CA IX antibody-conjugated, magnetic field-guided/activated nanoparticles as tools in anticancer strategies.