Sizing Single Cantilever Beam Specimens for Characterizing Facesheet/Core Peel Debonding in Sandwich Structure

NASA Technical Reports Server (NTRS)

Ratcliffe, James G.

2010-01-01

This paper details part of an effort focused on the development of a standardized facesheet/core peel debonding test procedure. The purpose of the test is to characterize facesheet/core peel in sandwich structure, accomplished through the measurement of the critical strain energy release rate associated with the debonding process. The specific test method selected for the standardized test procedure utilizes a single cantilever beam (SCB) specimen configuration. The objective of the current work is to develop a method for establishing SCB specimen dimensions. This is achieved by imposing specific limitations on specimen dimensions, with the objectives of promoting a linear elastic specimen response, and simplifying the data reduction method required for computing the critical strain energy release rate associated with debonding. The sizing method is also designed to be suitable for incorporation into a standardized test protocol. Preliminary application of the resulting sizing method yields practical specimen dimensions.

Switching industrial production processes from complex to defined media: method development and case study using the example of Penicillium chrysogenum.

PubMed

Posch, Andreas E; Spadiut, Oliver; Herwig, Christoph

2012-06-22

Filamentous fungi are versatile cell factories and widely used for the production of antibiotics, organic acids, enzymes and other industrially relevant compounds at large scale. As a fact, industrial production processes employing filamentous fungi are commonly based on complex raw materials. However, considerable lot-to-lot variability of complex media ingredients not only demands for exhaustive incoming components inspection and quality control, but unavoidably affects process stability and performance. Thus, switching bioprocesses from complex to defined media is highly desirable. This study presents a strategy for strain characterization of filamentous fungi on partly complex media using redundant mass balancing techniques. Applying the suggested method, interdependencies between specific biomass and side-product formation rates, production of fructooligosaccharides, specific complex media component uptake rates and fungal strains were revealed. A 2-fold increase of the overall penicillin space time yield and a 3-fold increase in the maximum specific penicillin formation rate were reached in defined media compared to complex media. The newly developed methodology enabled fast characterization of two different industrial Penicillium chrysogenum candidate strains on complex media based on specific complex media component uptake kinetics and identification of the most promising strain for switching the process from complex to defined conditions. Characterization at different complex/defined media ratios using only a limited number of analytical methods allowed maximizing the overall industrial objectives of increasing both, method throughput and the generation of scientific process understanding.

Switching industrial production processes from complex to defined media: method development and case study using the example of Penicillium chrysogenum

PubMed Central

2012-01-01

Background Filamentous fungi are versatile cell factories and widely used for the production of antibiotics, organic acids, enzymes and other industrially relevant compounds at large scale. As a fact, industrial production processes employing filamentous fungi are commonly based on complex raw materials. However, considerable lot-to-lot variability of complex media ingredients not only demands for exhaustive incoming components inspection and quality control, but unavoidably affects process stability and performance. Thus, switching bioprocesses from complex to defined media is highly desirable. Results This study presents a strategy for strain characterization of filamentous fungi on partly complex media using redundant mass balancing techniques. Applying the suggested method, interdependencies between specific biomass and side-product formation rates, production of fructooligosaccharides, specific complex media component uptake rates and fungal strains were revealed. A 2-fold increase of the overall penicillin space time yield and a 3-fold increase in the maximum specific penicillin formation rate were reached in defined media compared to complex media. Conclusions The newly developed methodology enabled fast characterization of two different industrial Penicillium chrysogenum candidate strains on complex media based on specific complex media component uptake kinetics and identification of the most promising strain for switching the process from complex to defined conditions. Characterization at different complex/defined media ratios using only a limited number of analytical methods allowed maximizing the overall industrial objectives of increasing both, method throughput and the generation of scientific process understanding. PMID:22727013

[Results of a study of the diagnostic qualities of brucellosis and tularemic antigenic erythrocytic diagnostica].

PubMed

Tsybin, B P; Taran, I F; Tinker, A I

1975-09-01

The authors elaborated methods of preparation of brucella and tularemia antigenic erythrocytic diagnostic agents which were characterized as highly specific, specific and stable preparations in mass examination of humans and animals at various stages of the vaccinal and infectous processes. The simplicity of obtaining specific antigens intended for the sensitization of formalinized erythrocytes and stability of the results of results of reproduction of the methods of preparation of the antigenic erythrocytic diagnostic agents offered a possibility of recommending the mentioned methods of industrial preparation of the diagnostic agents.

WIPP waste characterization program sampling and analysis guidance manual

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

Not Available

1991-01-01

The Waste Isolation Pilot Plant (WIPP) Waste Characterization Program Sampling and Analysis Guidance Manual (Guidance Manual) provides a unified source of information on the sampling and analytical techniques that enable Department of Energy (DOE) facilities to comply with the requirements established in the current revision of the Quality Assurance Program Plan (QAPP) for the WIPP Experimental-Waste Characterization Program (the Program). This Guidance Manual includes all of the sampling and testing methodologies accepted by the WIPP Project Office (DOE/WPO) for use in implementing the Program requirements specified in the QAPP. This includes methods for characterizing representative samples of transuranic (TRU) wastesmore » at DOE generator sites with respect to the gas generation controlling variables defined in the WIPP bin-scale and alcove test plans, as well as waste container headspace gas sampling and analytical procedures to support waste characterization requirements under the WIPP test program and the Resource Conservation and Recovery Act (RCRA). The procedures in this Guidance Manual are comprehensive and detailed and are designed to provide the necessary guidance for the preparation of site specific procedures. The use of these procedures is intended to provide the necessary sensitivity, specificity, precision, and comparability of analyses and test results. The solutions to achieving specific program objectives will depend upon facility constraints, compliance with DOE Orders and DOE facilities' operating contractor requirements, and the knowledge and experience of the TRU waste handlers and analysts. With some analytical methods, such as gas chromatography/mass spectrometry, the Guidance Manual procedures may be used directly. With other methods, such as nondestructive/destructive characterization, the Guidance Manual provides guidance rather than a step-by-step procedure.« less

Characterization and identification of ubiquitin conjugation sites with E3 ligase recognition specificities.

PubMed

Nguyen, Van-Nui; Huang, Kai-Yao; Huang, Chien-Hsun; Chang, Tzu-Hao; Bretaña, Neil; Lai, K; Weng, Julia; Lee, Tzong-Yi

2015-01-01

In eukaryotes, ubiquitin-conjugation is an important mechanism underlying proteasome-mediated degradation of proteins, and as such, plays an essential role in the regulation of many cellular processes. In the ubiquitin-proteasome pathway, E3 ligases play important roles by recognizing a specific protein substrate and catalyzing the attachment of ubiquitin to a lysine (K) residue. As more and more experimental data on ubiquitin conjugation sites become available, it becomes possible to develop prediction models that can be scaled to big data. However, no development that focuses on the investigation of ubiquitinated substrate specificities has existed. Herein, we present an approach that exploits an iteratively statistical method to identify ubiquitin conjugation sites with substrate site specificities. In this investigation, totally 6259 experimentally validated ubiquitinated proteins were obtained from dbPTM. After having filtered out homologous fragments with 40% sequence identity, the training data set contained 2658 ubiquitination sites (positive data) and 5532 non-ubiquitinated sites (negative data). Due to the difficulty in characterizing the substrate site specificities of E3 ligases by conventional sequence logo analysis, a recursively statistical method has been applied to obtain significant conserved motifs. The profile hidden Markov model (profile HMM) was adopted to construct the predictive models learned from the identified substrate motifs. A five-fold cross validation was then used to evaluate the predictive model, achieving sensitivity, specificity, and accuracy of 73.07%, 65.46%, and 67.93%, respectively. Additionally, an independent testing set, completely blind to the training data of the predictive model, was used to demonstrate that the proposed method could provide a promising accuracy (76.13%) and outperform other ubiquitination site prediction tool. A case study demonstrated the effectiveness of the characterized substrate motifs for identifying ubiquitination sites. The proposed method presents a practical means of preliminary analysis and greatly diminishes the total number of potential targets required for further experimental confirmation. This method may help unravel their mechanisms and roles in E3 recognition and ubiquitin-mediated protein degradation.

Comparative advantages and limitations of the basic metrology methods applied to the characterization of nanomaterials.

PubMed

Linkov, Pavel; Artemyev, Mikhail; Efimov, Anton E; Nabiev, Igor

2013-10-07

Fabrication of modern nanomaterials and nanostructures with specific functional properties is both scientifically promising and commercially profitable. The preparation and use of nanomaterials require adequate methods for the control and characterization of their size, shape, chemical composition, crystalline structure, energy levels, pathways and dynamics of physical and chemical processes during their fabrication and further use. In this review, we discuss different instrumental methods for the analysis and metrology of materials and evaluate their advantages and limitations at the nanolevel.

High-Throughput and Cost-Effective Characterization of Induced Pluripotent Stem Cells.

PubMed

D'Antonio, Matteo; Woodruff, Grace; Nathanson, Jason L; D'Antonio-Chronowska, Agnieszka; Arias, Angelo; Matsui, Hiroko; Williams, Roy; Herrera, Cheryl; Reyna, Sol M; Yeo, Gene W; Goldstein, Lawrence S B; Panopoulos, Athanasia D; Frazer, Kelly A

2017-04-11

Reprogramming somatic cells to induced pluripotent stem cells (iPSCs) offers the possibility of studying the molecular mechanisms underlying human diseases in cell types difficult to extract from living patients, such as neurons and cardiomyocytes. To date, studies have been published that use small panels of iPSC-derived cell lines to study monogenic diseases. However, to study complex diseases, where the genetic variation underlying the disorder is unknown, a sizable number of patient-specific iPSC lines and controls need to be generated. Currently the methods for deriving and characterizing iPSCs are time consuming, expensive, and, in some cases, descriptive but not quantitative. Here we set out to develop a set of simple methods that reduce cost and increase throughput in the characterization of iPSC lines. Specifically, we outline methods for high-throughput quantification of surface markers, gene expression analysis of in vitro differentiation potential, and evaluation of karyotype with markedly reduced cost. Published by Elsevier Inc.

Food powders flowability characterization: theory, methods, and applications.

PubMed

Juliano, Pablo; Barbosa-Cánovas, Gustavo V

2010-01-01

Characterization of food powders flowability is required for predicting powder flow from hoppers in small-scale systems such as vending machines or at the industrial scale from storage silos or bins dispensing into powder mixing systems or packaging machines. This review covers conventional and new methods used to measure flowability in food powders. The method developed by Jenike (1964) for determining hopper outlet diameter and hopper angle has become a standard for the design of bins and is regarded as a standard method to characterize flowability. Moreover, there are a number of shear cells that can be used to determine failure properties defined by Jenike's theory. Other classic methods (compression, angle of repose) and nonconventional methods (Hall flowmeter, Johanson Indicizer, Hosokawa powder tester, tensile strength tester, powder rheometer), used mainly for the characterization of food powder cohesiveness, are described. The effect of some factors preventing flow, such as water content, temperature, time consolidation, particle composition and size distribution, is summarized for the characterization of specific food powders with conventional and other methods. Whereas time-consuming standard methods established for hopper design provide flow properties, there is yet little comparative evidence demonstrating that other rapid methods may provide similar flow prediction.

Characterization and prediction of residues determining protein functional specificity.

PubMed

Capra, John A; Singh, Mona

2008-07-01

Within a homologous protein family, proteins may be grouped into subtypes that share specific functions that are not common to the entire family. Often, the amino acids present in a small number of sequence positions determine each protein's particular functional specificity. Knowledge of these specificity determining positions (SDPs) aids in protein function prediction, drug design and experimental analysis. A number of sequence-based computational methods have been introduced for identifying SDPs; however, their further development and evaluation have been hindered by the limited number of known experimentally determined SDPs. We combine several bioinformatics resources to automate a process, typically undertaken manually, to build a dataset of SDPs. The resulting large dataset, which consists of SDPs in enzymes, enables us to characterize SDPs in terms of their physicochemical and evolutionary properties. It also facilitates the large-scale evaluation of sequence-based SDP prediction methods. We present a simple sequence-based SDP prediction method, GroupSim, and show that, surprisingly, it is competitive with a representative set of current methods. We also describe ConsWin, a heuristic that considers sequence conservation of neighboring amino acids, and demonstrate that it improves the performance of all methods tested on our large dataset of enzyme SDPs. Datasets and GroupSim code are available online at http://compbio.cs.princeton.edu/specificity/. Supplementary data are available at Bioinformatics online.

Evaluation of epididymal function through specific protein on spermatozoa.

PubMed

Del Río, A G; De Sánchez, L Z; Sirena, A

1984-01-01

Investigations were focused on the characterization of specific epididymal proteins on the human spermatozoa as a representative parameter for epididymal function. An easy and attainable method, suitable for investigators and clinical use, is proposed in this article.

Physical interpretation and development of ultrasonic nondestructive evaluation techniques applied to the quantitative characterization of textile composite materials

NASA Technical Reports Server (NTRS)

Miller, James G.

1993-01-01

In this Progress Report, we describe our current research activities concerning the development and implementation of advanced ultrasonic nondestructive evaluation methods applied to the characterization of stitched composite materials and bonded aluminum plate specimens. One purpose of this investigation is to identify and characterize specific features of polar backscatter interrogation which enhance the ability of ultrasound to detect flaws in a stitched composite laminate. Another focus is to explore the feasibility of implementing medical linear array imaging technology as a viable ultrasonic-based nondestructive evaluation method to inspect and characterize bonded aluminum lap joints. As an approach to implementing quantitative ultrasonic inspection methods to both of these materials, we focus on the physics that underlies the detection of flaws in such materials.

Oromucosal film preparations: classification and characterization methods.

PubMed

Preis, Maren; Woertz, Christina; Kleinebudde, Peter; Breitkreutz, Jörg

2013-09-01

Recently, the regulatory authorities have enlarged the variety of 'oromucosal preparations' by buccal films and orodispersible films. Various film preparations have entered the market and pharmacopoeias. Due to the novelty of the official monographs, no standardized characterization methods and quality specifications are included. This review reports the methods of choice to characterize oromucosal film preparations with respect to biorelevant characterization and quality control. Commonly used dissolution tests for other dosage forms are not transferable for films in all cases. Alternatives and guidance on decision, which methods are favorable for film preparations are discussed. Furthermore, issues about requirements for film dosage forms are reflected. Oromucosal film preparations offer a wide spectrum of opportunities. There are a lot of suggestions in the literature on how to control the quality of these innovative products, but no standardized tests are available. Regulatory authorities need to define the standards and quality requirements more precisely.

New method for evaluating irreversible adsorption and stationary phase bleed in gas chromatographic capillary columns.

PubMed

Wright, Bob W; Wright, Cherylyn W

2012-10-26

A novel method is described for the evaluation of irreversible adsorption and column bleed in gas chromatographic (GC) columns using a tandem GC approach. This work specifically determined the degree of irreversible adsorption behavior of specific sulfur and phosphorous containing test probe compounds at levels ranging from approximately 50 picograms (pg) to 1 nanogram (ng) on selected gas chromatographic columns. This method does not replace existing evaluation methods that characterize reversible adsorption but provides an additional tool. The test compounds were selected due to their ease of adsorption and their importance in the specific trace analytical detection methodology being developed. Replicate chromatographic columns with 5% phenylmethylpolysiloxane (PMS), polyethylene glycol (wax), trifluoropropylpolysiloxane (TFP), or 78% cyanopropylpolysiloxane stationary phases from a variety of vendors were evaluated. As expected, the results demonstrate that the different chromatographic phases exhibit differing degrees of irreversible adsorption behavior. The results also indicate that all manufacturers do not produce equally inert columns nor are columns from a given manufacturer identical. The wax-coated columns for the test probes used were more inert as a group than 5% PMS coated columns, and they were more reproducibly manufactured. Both TFP and 78% cyanopropylpolysiloxane columns displayed superior inertness to the test compounds compared to either 5% PMS- or wax-coated columns. Irreversible adsorption behavior was characterized for a limited range of stationary phase film thicknesses. In addition, the method was shown effective for characterizing column bleed and methods to remove bleed components. This method is useful in screening columns for demanding applications and to obtain diagnostic information related to improved preparation methods. Copyright © 2012 Elsevier B.V. All rights reserved.

Bacteriophage Amplification-Coupled Detection and Identification of Bacterial Pathogens

NASA Astrophysics Data System (ADS)

Cox, Christopher R.; Voorhees, Kent J.

Current methods of species-specific bacterial detection and identification are complex, time-consuming, and often require expensive specialized equipment and highly trained personnel. Numerous biochemical and genotypic identification methods have been applied to bacterial characterization, but all rely on tedious microbiological culturing practices and/or costly sequencing protocols which render them impractical for deployment as rapid, cost-effective point-of-care or field detection and identification methods. With a view towards addressing these shortcomings, we have exploited the evolutionarily conserved interactions between a bacteriophage (phage) and its bacterial host to develop species-specific detection methods. Phage amplification-coupled matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) was utilized to rapidly detect phage propagation resulting from species-specific in vitro bacterial infection. This novel signal amplification method allowed for bacterial detection and identification in as little as 2 h, and when combined with disulfide bond reduction methods developed in our laboratory to enhance MALDI-TOF-MS resolution, was observed to lower the limit of detection by several orders of magnitude over conventional spectroscopy and phage typing methods. Phage amplification has been combined with lateral flow immunochromatography (LFI) to develop rapid, easy-to-operate, portable, species-specific point-of-care (POC) detection devices. Prototype LFI detectors have been developed and characterized for Yersinia pestis and Bacillus anthracis, the etiologic agents of plague and anthrax, respectively. Comparable sensitivity and rapidity was observed when phage amplification was adapted to a species-specific handheld LFI detector, thus allowing for rapid, simple, POC bacterial detection and identification while eliminating the need for bacterial culturing or DNA isolation and amplification techniques.

Linear polarizer local characterizations by polarimetric imaging for applications to polarimetric sensors for torque measurement for hybrid cars

NASA Astrophysics Data System (ADS)

Georges, F.; Remouche, M.; Meyrueis, P.

2011-06-01

Usually manufacturer's specifications do not deal with the ability of linear sheet polarizers to have a constant transmittance function over their geometric area. These parameters are fundamental for developing low cost polarimetric sensors(for instance rotation, torque, displacement) specifically for hybrid car (thermic + electricity power). It is then necessary to specially characterize commercial polarizers sheets to find if they are adapted to this kind of applications. In this paper, we present measuring methods and bench developed for this purpose, and some preliminary characterization results. We state conclusions for effective applications to hybrid car gearbox control and monitoring.

APPLICATIONS OF BOREHOLE-ACOUSTIC METHODS IN ROCK MECHANICS.

USGS Publications Warehouse

Paillet, Frederick L.

1985-01-01

Acoustic-logging methods using a considerable range of wavelengths and frequencies have proven very useful in the in situ characterization of deeply buried crystalline rocks. Seismic velocities are useful in investigating the moduli of unfractured rock, and in producing a continuous record of rock quality for comparison with discontinuous intervals of core. The considerable range of frequencies makes the investigation of scale effects possible in both fractured and unfractured rock. Several specific methods for the characterization of in situ permeability have been developed and verified in the field.

NetMHCIIpan-2.0 - Improved pan-specific HLA-DR predictions using a novel concurrent alignment and weight optimization training procedure.

PubMed

Nielsen, Morten; Justesen, Sune; Lund, Ole; Lundegaard, Claus; Buus, Søren

2010-11-13

Binding of peptides to Major Histocompatibility class II (MHC-II) molecules play a central role in governing responses of the adaptive immune system. MHC-II molecules sample peptides from the extracellular space allowing the immune system to detect the presence of foreign microbes from this compartment. Predicting which peptides bind to an MHC-II molecule is therefore of pivotal importance for understanding the immune response and its effect on host-pathogen interactions. The experimental cost associated with characterizing the binding motif of an MHC-II molecule is significant and large efforts have therefore been placed in developing accurate computer methods capable of predicting this binding event. Prediction of peptide binding to MHC-II is complicated by the open binding cleft of the MHC-II molecule, allowing binding of peptides extending out of the binding groove. Moreover, the genes encoding the MHC molecules are immensely diverse leading to a large set of different MHC molecules each potentially binding a unique set of peptides. Characterizing each MHC-II molecule using peptide-screening binding assays is hence not a viable option. Here, we present an MHC-II binding prediction algorithm aiming at dealing with these challenges. The method is a pan-specific version of the earlier published allele-specific NN-align algorithm and does not require any pre-alignment of the input data. This allows the method to benefit also from information from alleles covered by limited binding data. The method is evaluated on a large and diverse set of benchmark data, and is shown to significantly out-perform state-of-the-art MHC-II prediction methods. In particular, the method is found to boost the performance for alleles characterized by limited binding data where conventional allele-specific methods tend to achieve poor prediction accuracy. The method thus shows great potential for efficient boosting the accuracy of MHC-II binding prediction, as accurate predictions can be obtained for novel alleles at highly reduced experimental costs. Pan-specific binding predictions can be obtained for all alleles with know protein sequence and the method can benefit by including data in the training from alleles even where only few binders are known. The method and benchmark data are available at http://www.cbs.dtu.dk/services/NetMHCIIpan-2.0.

Isolation and characterization of living circulating tumor cells in patients by immunomagnetic negative enrichment coupled with flow cytometry.

PubMed

Lu, Yusheng; Liang, Haiyan; Yu, Ting; Xie, Jingjing; Chen, Shuming; Dong, Haiyan; Sinko, Patrick J; Lian, Shu; Xu, Jianguo; Wang, Jichuang; Yu, Suhong; Shao, Jingwei; Yuan, Bo; Wang, Lie; Jia, Lee

2015-09-01

This study was aimed at establishing a sensitive and specific isolation, characterization, and enumeration method for living circulating tumor cells (CTCs) in patients with colorectal carcinoma. Quantitative isolation and characterization of CTCs were performed through a combination of immunomagnetic negative enrichment and fluorescence-activated cell sorting. Isolated CTCs were identified by immunofluorescence staining. The viability and purity of the sorted cells were determined by flow cytometry. Blood samples spiked with HCT116 cells (range, 3-250 cells) were used to determine specificity, recovery, and sensitivity. The method was used to enumerate, characterize, and isolate living CTCs in 10 mL of blood from patients with colorectal carcinoma. The average recovery of HCT116 cells was 61% or more at each spiking level, and the correlation coefficient was 0.992. An analysis of samples from all 18 patients with colorectal carcinoma revealed that 94.4% were positive for CTCs with an average of 33 ± 24 CTCs per 10 mL of blood and with a diameter of 14 to 20 μm (vs 8-12 μm for lymphoma). All patients were CD47(+) , with only 4.3% to 61.2% being CD44(+) . The number of CTCs was well correlated with the patient TNM stage and could be detected in patients at an early cancer stage. The sorted cells could be recultured, and their viability was preserved. This method provides a novel technique for highly sensitive and specific detection and isolation of CTCs in patients with colorectal carcinoma. This method complements the existing approaches for the de novo functional identification of a wide variety of CTC types. It is likely to help in predicting a patient's disease progression and potentially in selecting the appropriate treatment. © 2015 American Cancer Society.

Analysis and feasibility of asphalt pavement performance-based specifications for WisDOT.

DOT National Transportation Integrated Search

2016-12-25

Literature review of most recent methods used for effective characterization of asphalt mixtures resulted in selecting aset of test methods for measuring mixture resistance for rutting and moisture damage at high temperature, fatigue cracking at inte...

Aerosol generation and characterization of multi-walled carbon nanotubes exposed to cells cultured at the air-liquid interface.

PubMed

Polk, William W; Sharma, Monita; Sayes, Christie M; Hotchkiss, Jon A; Clippinger, Amy J

2016-04-23

Aerosol generation and characterization are critical components in the assessment of the inhalation hazards of engineered nanomaterials (NMs). An extensive review was conducted on aerosol generation and exposure apparatus as part of an international expert workshop convened to discuss the design of an in vitro testing strategy to assess pulmonary toxicity following exposure to aerosolized particles. More specifically, this workshop focused on the design of an in vitro method to predict the development of pulmonary fibrosis in humans following exposure to multi-walled carbon nanotubes (MWCNTs). Aerosol generators, for dry or liquid particle suspension aerosolization, and exposure chambers, including both commercially available systems and those developed by independent researchers, were evaluated. Additionally, characterization methods that can be used and the time points at which characterization can be conducted in order to interpret in vitro exposure results were assessed. Summarized below is the information presented and discussed regarding the relevance of various aerosol generation and characterization techniques specific to aerosolized MWCNTs exposed to cells cultured at the air-liquid interface (ALI). The generation of MWCNT aerosols relevant to human exposures and their characterization throughout exposure in an ALI system is critical for extrapolation of in vitro results to toxicological outcomes in humans.

Serological purification of polysaccharide antigens from Streptococcus mutans serotypes a and d: characterization of multiple antigenic determinants.

PubMed

Linzer, R; Mukasa, H; Slade, H D

1975-10-01

The polysaccharide antigen preparations from serotype a and serotype d strains of Streptococcus mutans contained both a serotype-specific antigenic determinant and a common a-d antigenic determinant, as demonstrated by agar gel diffusion studies and a quantitative cross-precipitin assay. The chromatographically purified antigens were isolated by a method which depended on their serological specificity to determine if these two antigenic determinants were located on the same molecule. The a and d polysaccharides were recovered from specific antigen-antibody complexes and characterized with respect to their immunological specificity and chemical composition. Agar gel diffusion tests demonstrated that, in both the a and d preparations, the serotype-specific antigenic determinant and the common a-d antigenic determinant were present in one molecule.

Preparation and Evaluation of Multiple Nanoemulsions Containing Gadolinium (III) Chelate as a Potential Magnetic Resonance Imaging (MRI) Contrast Agent.

PubMed

Sigward, Estelle; Corvis, Yohann; Doan, Bich-Thuy; Kindsiko, Kadri; Seguin, Johanne; Scherman, Daniel; Brossard, Denis; Mignet, Nathalie; Espeau, Philippe; Crauste-Manciet, Sylvie

2015-09-01

The objective was to develop, characterize and assess the potentiality of W1/O/W2 self-emulsifying multiple nanoemulsions to enhance signal/noise ratio for Magnetic Resonance Imaging (MRI). For this purpose, a new formulation, was designed for encapsulation efficiency and stability. Various methods were used to characterize encapsulation efficiency ,in particular calorimetric methods (Differential Scanning Calorimetry (DSC), thermogravimetry analysis) and ultrafiltration. MRI in vitro relaxivities were assessed on loaded DTPA-Gd multiple nanoemulsions. Characterization of the formulation, in particular of encapsulation efficiency was a challenge due to interactions found with ultrafiltration method. Thanks to the specifically developed DSC protocol, we were able to confirm the formation of multiple nanoemulsions, differentiate loaded from unloaded nanoemulsions and measure the encapsulation efficiency which was found to be quite high with a 68% of drug loaded. Relaxivity studies showed that the self-emulsifying W/O/W nanoemulsions were positive contrast agents, exhibiting higher relaxivities than those of the DTPA-Gd solution taken as a reference. New self-emulsifying multiple nanoemulsions that were able to load satisfactory amounts of contrasting agent were successfully developed as potential MRI contrasting agents. A specific DSC protocol was needed to be developed to characterize these complex systems as it would be useful to develop these self-formation formulations.

Characterization of lipid-rich plaques using spectroscopic optical coherence tomography

NASA Astrophysics Data System (ADS)

Nam, Hyeong Soo; Song, Joon Woo; Jang, Sun-Joo; Lee, Jae Joong; Oh, Wang-Yuhl; Kim, Jin Won; Yoo, Hongki

2016-07-01

Intravascular optical coherence tomography (IV-OCT) is a high-resolution imaging method used to visualize the internal structures of walls of coronary arteries in vivo. However, accurate characterization of atherosclerotic plaques with gray-scale IV-OCT images is often limited by various intrinsic artifacts. In this study, we present an algorithm for characterizing lipid-rich plaques with a spectroscopic OCT technique based on a Gaussian center of mass (GCOM) metric. The GCOM metric, which reflects the absorbance properties of lipids, was validated using a lipid phantom. In addition, the proposed characterization method was successfully demonstrated in vivo using an atherosclerotic rabbit model and was found to have a sensitivity and specificity of 94.3% and 76.7% for lipid classification, respectively.

Detection of nucleic acids by multiple sequential invasive cleavages

DOEpatents

Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann D.

1999-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

Nucleic acid detection kits

DOEpatents

Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann; Kwiatkowski, Robert W.; Vavra, Stephanie H.

2005-03-29

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of nucleic acid from various viruses in a sample.

Detection of nucleic acids by multiple sequential invasive cleavages 02

DOEpatents

Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann D.

2002-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

Detection of nucleic acids by multiple sequential invasive cleavages

DOEpatents

Hall, Jeff G; Lyamichev, Victor I; Mast, Andrea L; Brow, Mary Ann D

2012-10-16

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

Thickness-Independent Ultrasonic Imaging Applied to Abrasive Cut-Off Wheels: An Advanced Aerospace Materials Characterization Method for the Abrasives Industry. A NASA Lewis Research Center Technology Transfer Case History

NASA Technical Reports Server (NTRS)

Roth, Don J.; Farmer, Donald A.

1998-01-01

Abrasive cut-off wheels are at times unintentionally manufactured with nonuniformity that is difficult to identify and sufficiently characterize without time-consuming, destructive examination. One particular nonuniformity is a density variation condition occurring around the wheel circumference or along the radius, or both. This density variation, depending on its severity, can cause wheel warpage and wheel vibration resulting in unacceptable performance and perhaps premature failure of the wheel. Conventional nondestructive evaluation methods such as ultrasonic c-scan imaging and film radiography are inaccurate in their attempts at characterizing the density variation because a superimposing thickness variation exists as well in the wheel. In this article, the single transducer thickness-independent ultrasonic imaging method, developed specifically to allow more accurate characterization of aerospace components, is shown to precisely characterize the extent of the density variation in a cut-off wheel having a superimposing thickness variation. The method thereby has potential as an effective quality control tool in the abrasives industry for the wheel manufacturer.

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.

Multiscale Structure of UXO Site Characterization: Spatial Estimation and Uncertainty Quantification

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

Ostrouchov, George; Doll, William E.; Beard, Les P.

2009-01-01

Unexploded ordnance (UXO) site characterization must consider both how the contamination is generated and how we observe that contamination. Within the generation and observation processes, dependence structures can be exploited at multiple scales. We describe a conceptual site characterization process, the dependence structures available at several scales, and consider their statistical estimation aspects. It is evident that most of the statistical methods that are needed to address the estimation problems are known but their application-specific implementation may not be available. We demonstrate estimation at one scale and propose a representation for site contamination intensity that takes full account of uncertainty,more » is flexible enough to answer regulatory requirements, and is a practical tool for managing detailed spatial site characterization and remediation. The representation is based on point process spatial estimation methods that require modern computational resources for practical application. These methods have provisions for including prior and covariate information.« less

Methods And Devices For Characterizing Duplex Nucleic Acid Molecules

DOEpatents

Akeson, Mark; Vercoutere, Wenonah; Haussler, David; Winters-Hilt, Stephen

2005-08-30

Methods and devices are provided for characterizing a duplex nucleic acid, e.g., a duplex DNA molecule. In the subject methods, a fluid conducting medium that includes a duplex nucleic acid molecule is contacted with a nanopore under the influence of an applied electric field and the resulting changes in current through the nanopore caused by the duplex nucleic acid molecule are monitored. The observed changes in current through the nanopore are then employed as a set of data values to characterize the duplex nucleic acid, where the set of data values may be employed in raw form or manipulated, e.g., into a current blockade profile. Also provided are nanopore devices for practicing the subject methods, where the subject nanopore devices are characterized by the presence of an algorithm which directs a processing means to employ monitored changes in current through a nanopore to characterize a duplex nucleic acid molecule responsible for the current changes. The subject methods and devices find use in a variety of applications, including, among other applications, the identification of an analyte duplex DNA molecule in a sample, the specific base sequence at a single nulceotide polymorphism (SNP), and the sequencing of duplex DNA molecules.

Method for the evaluation 3D noncontact inspection systems

NASA Astrophysics Data System (ADS)

Harding, Kevin

2011-08-01

Three dimensional, optical measurement systems are becoming more widely used in applications ranging from aerospace to automotive. These systems offer the potential for high speed, good accuracy, and more complete information than older contact based technology. However, the primary standards employed by many to evaluate these systems were specifically designed around touch probe based coordinate measurement machines (CMMs). These standards were designed to work with the limitations of touch probes, and in many cases cannot measure the types of features and errors associated with non-contact systems. This paper will discuss the deficiencies of employing contact based characterization tests to non-contact systems, and suggest a new set of tests specifically to cover the many aspects pertinent to non-contact, optical 3D measurement systems. Some of the performance aspects addressed in this characterization method include: sensitivity to surface reflectivity and roughness, the effect of angle of incidence of measurements, means to characterize volumetric variations that may fit complex functions, and considerations of both spatial and depth resolutions. Specific application areas will be discussed as well as the use of artifacts to provide practical functional data that can predict system performance on real world parts.

Optimal mask characterization by Surrogate Wafer Print (SWaP) method

NASA Astrophysics Data System (ADS)

Kimmel, Kurt R.; Hoellein, Ingo; Peters, Jan Hendrick; Ackmann, Paul; Connolly, Brid; West, Craig

2008-10-01

Traditionally, definition of mask specifications is done completely by the mask user, while characterization of the mask relative to the specifications is done completely by the mask maker. As the challenges of low-k1 imaging continue to grow in scope of designs and in absolute complexity, the inevitable partnership between wafer lithographers and mask makers has strengthened as well. This is reflected in the jointly owned mask facilities and device manufacturers' continued maintenance of fully captive mask shops which foster the closer mask-litho relationships. However, while some device manufacturers have leveraged this to optimize mask specifications before the mask is built and, therefore, improve mask yield and cost, the opportunity for post-fabrication partnering on mask characterization is more apparent and compelling. The Advanced Mask Technology Center (AMTC) has been investigating the concept of assessing how a mask images, rather than the mask's physical attributes, as a technically superior and lower-cost method to characterize a mask. The idea of printing a mask under its intended imaging conditions, then characterizing the imaged wafer as a surrogate for traditional mask inspections and measurements represents the ultimate method to characterize a mask's performance, which is most meaningful to the user. Surrogate wafer print (SWaP) is already done as part of leading-edge wafer fab mask qualification to validate defect and dimensional performance. In the past, the prospect of executing this concept has generally been summarily discarded as technically untenable and logistically intractable. The AMTC published a paper at BACUS 2007 successfully demonstrating the performance of SWaP for the characterization of defects as an alternative to traditional mask inspection [1]. It showed that this concept is not only feasible, but, in some cases, desirable. This paper expands on last year's work at AMTC to assess the full implementation of SWaP as an enhancement to mask characterization quality including defectivity, dimensional control, pattern fidelity, and in-plane distortion. We present a thorough analysis of both the technical and logistical challenges coupled with an objective view of the advantages and disadvantages from both the technical and financial perspectives. The analysis and model used by the AMTC will serve to provoke other mask shops to prepare their own analyses then consider this new paradigm for mask characterization and qualification.

Species-Specific Thiol-Disulfide Equilibrium Constant: A Tool To Characterize Redox Transitions of Biological Importance.

PubMed

Mirzahosseini, Arash; Somlyay, Máté; Noszál, Béla

2015-08-13

Microscopic redox equilibrium constants, a new species-specific type of physicochemical parameters, were introduced and determined to quantify thiol-disulfide equilibria of biological significance. The thiol-disulfide redox equilibria of glutathione with cysteamine, cysteine, and homocysteine were approached from both sides, and the equilibrium mixtures were analyzed by quantitative NMR methods to characterize the highly composite, co-dependent acid-base and redox equilibria. The directly obtained, pH-dependent, conditional constants were then decomposed by a new evaluation method, resulting in pH-independent, microscopic redox equilibrium constants for the first time. The 80 different, microscopic redox equilibrium constant values show close correlation with the respective thiolate basicities and provide sound means for the development of potent agents against oxidative stress.

Isolation & characterization of Brucella melitensis isolated from patients suspected for human brucellosis in India

PubMed Central

Barua, Anita; Kumar, Ashu; Thavaselvam, Duraipandian; Mangalgi, Smita; Prakash, Archana; Tiwari, Sapana; Arora, Sonia; Sathyaseelan, Kannusamy

2016-01-01

Background & objectives: Brucellosis is endemic in the southern part of India. A combination of biochemical, serological and molecular methods is required for identification and biotyping of Brucella. The present study describes the isolation and biochemical, molecular characterization of Brucella melitensis from patients suspected for human brucellosis. Methods: The blood samples were collected from febrile patients suspected to have brucellosis. A total of 18 isolates were obtained from 102 blood samples subjected to culture. The characterization of these 18 isolates was done by growth on Brucella specific medium, biochemical reactions, CO2 requirement, H2S production, agglutination with A and M mono-specific antiserum, dye sensitivity to basic fuchsin and thionin. Further, molecular characterization of the isolates was done by amplification of B. melitensis species specific IS711 repetitive DNA fragment and 16S (rRNA) sequence analysis. PCR-restriction fragment length polymorphism (RFLP) analysis of omp2 locus and IS711 gene was also done for molecular characterization. Results: All 102 suspected samples were subjected to bacteria isolation and of these, 18 isolates could be recovered on blood culture. The biochemical, PCR and PCR-RFLP and 16s rRNA sequencing revealed that all isolates were of B. melitensis and matched exactly with reference strain B. melitensis 16M. Interpretation & conclusions: The present study showed an overall isolation rate of 17.64 per cent for B. melitensis. There is a need to establish facilities for isolation and characterization of Brucella species for effective clinical management of the disease among patients as well as surveillance and control of infection in domestic animals. Further studies are needed from different geographical areas of the country with different level of endemicity to plan and execute control strategies against human brucellosis. PMID:27488010

Detection, Characterization, and Typing of Shiga Toxin-Producing Escherichia coli.

PubMed

Parsons, Brendon D; Zelyas, Nathan; Berenger, Byron M; Chui, Linda

2016-01-01

Shiga toxin-producing Escherichia coli (STEC) are responsible for gastrointestinal diseases reported in numerous outbreaks around the world. Given the public health importance of STEC, effective detection, characterization and typing is critical to any medical laboratory system. While non-O157 serotypes account for the majority of STEC infections, frontline microbiology laboratories may only screen for STEC using O157-specific agar-based methods. As a result, non-O157 STEC infections are significantly under-reported. This review discusses recent advances on the detection, characterization and typing of STEC with emphasis on work performed at the Alberta Provincial Laboratory for Public Health (ProvLab). Candidates for the detection of all STEC serotypes include chromogenic agars, enzyme immunoassays (EIA) and quantitative real time polymerase chain reaction (qPCR). Culture methods allow further characterization of isolates, whereas qPCR provides the greatest sensitivity and specificity, followed by EIA. The virulence gene profiles using PCR arrays and stx gene subtypes can subsequently be determined. Different non-O157 serotypes exhibit markedly different virulence gene profiles and a greater prevalence of stx1 than stx2 subtypes compared to O157:H7 isolates. Finally, recent innovations in whole genome sequencing (WGS) have allowed it to emerge as a candidate for the characterization and typing of STEC in diagnostic surveillance isolates. Methods of whole genome analysis such as single nucleotide polymorphisms and k-mer analysis are concordant with epidemiological data and standard typing methods, such as pulsed-field gel electrophoresis and multiple-locus variable number tandem repeat analysis while offering additional strain differentiation. Together these findings highlight improved strategies for STEC detection using currently available systems and the development of novel approaches for future surveillance.

Detection, Characterization, and Typing of Shiga Toxin-Producing Escherichia coli

PubMed Central

Parsons, Brendon D.; Zelyas, Nathan; Berenger, Byron M.; Chui, Linda

2016-01-01

Shiga toxin-producing Escherichia coli (STEC) are responsible for gastrointestinal diseases reported in numerous outbreaks around the world. Given the public health importance of STEC, effective detection, characterization and typing is critical to any medical laboratory system. While non-O157 serotypes account for the majority of STEC infections, frontline microbiology laboratories may only screen for STEC using O157-specific agar-based methods. As a result, non-O157 STEC infections are significantly under-reported. This review discusses recent advances on the detection, characterization and typing of STEC with emphasis on work performed at the Alberta Provincial Laboratory for Public Health (ProvLab). Candidates for the detection of all STEC serotypes include chromogenic agars, enzyme immunoassays (EIA) and quantitative real time polymerase chain reaction (qPCR). Culture methods allow further characterization of isolates, whereas qPCR provides the greatest sensitivity and specificity, followed by EIA. The virulence gene profiles using PCR arrays and stx gene subtypes can subsequently be determined. Different non-O157 serotypes exhibit markedly different virulence gene profiles and a greater prevalence of stx1 than stx2 subtypes compared to O157:H7 isolates. Finally, recent innovations in whole genome sequencing (WGS) have allowed it to emerge as a candidate for the characterization and typing of STEC in diagnostic surveillance isolates. Methods of whole genome analysis such as single nucleotide polymorphisms and k-mer analysis are concordant with epidemiological data and standard typing methods, such as pulsed-field gel electrophoresis and multiple-locus variable number tandem repeat analysis while offering additional strain differentiation. Together these findings highlight improved strategies for STEC detection using currently available systems and the development of novel approaches for future surveillance. PMID:27148176

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.

An overview of the characterization of occupational exposure to nanoaerosols in workplaces

NASA Astrophysics Data System (ADS)

Castellano, Paola; Ferrante, Riccardo; Curini, Roberta; Canepari, Silvia

2009-05-01

Currently, there is a lack of standardized sampling and metric methods that can be applied to measure the level of exposure to nanosized aerosols. Therefore, any attempt to characterize exposure to nanoparticles (NP) in a workplace must involve a multifaceted approach characterized by different sampling and analytical techniques to measure all relevant characteristics of NP exposure. Furthermore, as NP aerosols are always complex mixtures of multiple origins, sampling and analytical methods need to be improved to selectively evaluate the apportionment from specific sources to the final nanomaterials. An open question at the world's level is how to relate specific toxic effects of NP with one or more among several different parameters (such as particle size, mass, composition, surface area, number concentration, aggregation or agglomeration state, water solubility and surface chemistry). As the evaluation of occupational exposure to NP in workplaces needs dimensional and chemical characterization, the main problem is the choice of the sampling and dimensional separation techniques. Therefore a convenient approach to allow a satisfactory risk assessment could be the contemporary use of different sampling and measuring techniques for particles with known toxicity in selected workplaces. Despite the lack of specific NP exposure limit values, exposure metrics, appropriate to nanoaerosols, are discussed in the Technical Report ISO/TR 27628:2007 with the aim to enable occupational hygienists to characterize and monitor nanoaerosols in workplaces. Moreover, NIOSH has developed the Document Approaches to Safe Nanotechnology (intended to be an information exchange with NIOSH) in order to address current and future research needs to understanding the potential risks that nanotechnology may have to workers.

Methods for Instructional Diagnosis with Limited Available Resources.

ERIC Educational Resources Information Center

Gillmore, Gerald M.; Clark, D. Joseph

College teaching should be approached with the same careful delineation of problems and systematic attempts to find solutions which characterize research. Specific methods for the diagnosis of instructional problems include audio-video taping, use of teaching assistants, colleague assistance, classroom tests, student projects in and out of class,…

Methodology for the specification of communication activities within the framework of a multi-layered architecture: Toward the definition of a knowledge base

NASA Astrophysics Data System (ADS)

Amyay, Omar

A method defined in terms of synthesis and verification steps is presented. The specification of the services and protocols of communication within a multilayered architecture of the Open Systems Interconnection (OSI) type is an essential issue for the design of computer networks. The aim is to obtain an operational specification of the protocol service couple of a given layer. Planning synthesis and verification steps constitute a specification trajectory. The latter is based on the progressive integration of the 'initial data' constraints and verification of the specification originating from each synthesis step, through validity constraints that characterize an admissible solution. Two types of trajectories are proposed according to the style of the initial specification of the service protocol couple: operational type and service supplier viewpoint; knowledge property oriented type and service viewpoint. Synthesis and verification activities were developed and formalized in terms of labeled transition systems, temporal logic and epistemic logic. The originality of the second specification trajectory and the use of the epistemic logic are shown. An 'artificial intelligence' approach enables a conceptual model to be defined for a knowledge base system for implementing the method proposed. It is structured in three levels of representation of the knowledge relating to the domain, the reasoning characterizing synthesis and verification activities and the planning of the steps of a specification trajectory.

Rapid characterization of microorganisms by mass spectrometry--what can be learned and how?

PubMed

Fenselau, Catherine C

2013-08-01

Strategies for the rapid and reliable analysis of microorganisms have been sought to meet national needs in defense, homeland security, space exploration, food and water safety, and clinical diagnosis. Mass spectrometry has long been a candidate technique because it is extremely rapid and can provide highly specific information. It has excellent sensitivity. Molecular and fragment ion masses provide detailed fingerprints, which can also be interpreted. Mass spectrometry is also a broad band method--everything has a mass--and it is automatable. Mass spectrometry is a physiochemical method that is orthogonal and complementary to biochemical and morphological methods used to characterize microorganisms.

Capillary zone electrophoresis method for a highly glycosylated and sialylated recombinant protein: development, characterization and application for process development.

PubMed

Zhang, Le; Lawson, Ken; Yeung, Bernice; Wypych, Jette

2015-01-06

A purity method based on capillary zone electrophoresis (CZE) has been developed for the separation of isoforms of a highly glycosylated protein. The separation was found to be driven by the number of sialic acids attached to each isoform. The method has been characterized using orthogonal assays and shown to have excellent specificity, precision and accuracy. We have demonstrated the CZE method is a useful in-process assay to support cell culture and purification development of this glycoprotein. Compared to isoelectric focusing (IEF), the CZE method provides more quantitative results and higher sample throughput with excellent accuracy, qualities that are required for process development. In addition, the CZE method has been applied in the stability testing of purified glycoprotein samples.

Engineering peptide ligase specificity by proteomic identification of ligation sites.

PubMed

Weeks, Amy M; Wells, James A

2018-01-01

Enzyme-catalyzed peptide ligation is a powerful tool for site-specific protein bioconjugation, but stringent enzyme-substrate specificity limits its utility. We developed an approach for comprehensively characterizing peptide ligase specificity for N termini using proteome-derived peptide libraries. We used this strategy to characterize the ligation efficiency for >25,000 enzyme-substrate pairs in the context of the engineered peptide ligase subtiligase and identified a family of 72 mutant subtiligases with activity toward N-terminal sequences that were previously recalcitrant to modification. We applied these mutants individually for site-specific bioconjugation of purified proteins, including antibodies, and in algorithmically selected combinations for sequencing of the cellular N terminome with reduced sequence bias. We also developed a web application to enable algorithmic selection of the most efficient subtiligase variant(s) for bioconjugation to user-defined sequences. Our methods provide a new toolbox of enzymes for site-specific protein modification and a general approach for rapidly defining and engineering peptide ligase specificity.

EDITORIAL: (Nano)characterization of semiconductor materials and structures (Nano)characterization of semiconductor materials and structures

NASA Astrophysics Data System (ADS)

Bonanni, Alberta

2011-06-01

The latest impressive advancements in the epitaxial fabrication of semiconductors and in the refinement of characterization techniques have the potential to allow insight into the deep relation between materials' structural properties and their physical and chemical functionalities. Furthermore, while the comprehensive (nano)characterization of semiconductor materials and structures is becoming more and more necessary, a compendium of the currently available techniques is lacking. We are positive that an overview of the hurdles related to the specific methods, often leading to deceptive interpretations, will be most informative for the broad community working on semiconductors, and will help in shining some light onto a plethora of controversial reports found in the literature. From this perspective, with this special issue we address and highlight the challenges and misinterpretations related to complementary local (nanoscale) and more global experimental methods for the characterization of semiconductors. The six topical reviews and the three invited papers by leading experts in the specific fields collected in here are intended to provide the required broad overview on the possibilities of actual (nano)characterization methods, from the microscopy of single quantum structures, over the synchrotron-based absorption and diffraction of nano-objects, to the contentious detection of tiny magnetic signals by quantum interference and resonance techniques. We are grateful to all the authors for their valuable contributions. Moreover, I would like to thank the Editorial Board of the journal for supporting the realization of this special issue and for inviting me to serve as Guest Editor. We greatly appreciate the work of the reviewers, of the editorial staff of Semiconductor Science and Technology and of IOP Publishing. In particular, the efforts of Alice Malhador in coordinating this special issue are acknowledged.

Engineering entrainment and adaptation in limit cycle systems : From biological inspiration to applications in robotics.

PubMed

Buchli, Jonas; Righetti, Ludovic; Ijspeert, Auke Jan

2006-12-01

Periodic behavior is key to life and is observed in multiple instances and at multiple time scales in our metabolism, our natural environment, and our engineered environment. A natural way of modeling or generating periodic behavior is done by using oscillators, i.e., dynamical systems that exhibit limit cycle behavior. While there is extensive literature on methods to analyze such dynamical systems, much less work has been done on methods to synthesize an oscillator to exhibit some specific desired characteristics. The goal of this article is twofold: (1) to provide a framework for characterizing and designing oscillators and (2) to review how classes of well-known oscillators can be understood and related to this framework. The basis of the framework is to characterize oscillators in terms of their fundamental temporal and spatial behavior and in terms of properties that these two behaviors can be designed to exhibit. This focus on fundamental properties is important because it allows us to systematically compare a large variety of oscillators that might at first sight appear very different from each other. We identify several specifications that are useful for design, such as frequency-locking behavior, phase-locking behavior, and specific output signal shape. We also identify two classes of design methods by which these specifications can be met, namely offline methods and online methods. By relating these specifications to our framework and by presenting several examples of how oscillators have been designed in the literature, this article provides a useful methodology and toolbox for designing oscillators for a wide range of purposes. In particular, the focus on synthesis of limit cycle dynamical systems should be useful both for engineering and for computational modeling of physical or biological phenomena.

Characterization of Change and Significance for Clinical Findings in Radiology Reports Through Natural Language Processing.

PubMed

Hassanpour, Saeed; Bay, Graham; Langlotz, Curtis P

2017-06-01

We built a natural language processing (NLP) method to automatically extract clinical findings in radiology reports and characterize their level of change and significance according to a radiology-specific information model. We utilized a combination of machine learning and rule-based approaches for this purpose. Our method is unique in capturing different features and levels of abstractions at surface, entity, and discourse levels in text analysis. This combination has enabled us to recognize the underlying semantics of radiology report narratives for this task. We evaluated our method on radiology reports from four major healthcare organizations. Our evaluation showed the efficacy of our method in highlighting important changes (accuracy 99.2%, precision 96.3%, recall 93.5%, and F1 score 94.7%) and identifying significant observations (accuracy 75.8%, precision 75.2%, recall 75.7%, and F1 score 75.3%) to characterize radiology reports. This method can help clinicians quickly understand the key observations in radiology reports and facilitate clinical decision support, review prioritization, and disease surveillance.

Silicon cantilever functionalization for cellulose-specific chemical force imaging of switchgrass

DOE PAGES

Lee, Ida; Evans, Barbara R.; Foston, Marcus B.; ...

2015-05-08

A method for direct functionalization of silicon and silicon nitride cantilevers with bifunctional silanes was tested with model surfaces to determine adhesive forces for different hydrogen-bonding chemistries. Application for biomass surface characterization was tested by mapping switchgrass and isolated switchgrass cellulose in topographic and force-volume mode using a cellulose-specific cantilever.

A combined parasitological molecular approach for noninvasive characterization of parasitic nematode communities in wild hosts.

PubMed

Budischak, Sarah A; Hoberg, Eric P; Abrams, Art; Jolles, Anna E; Ezenwa, Vanessa O

2015-09-01

Most hosts are concurrently or sequentially infected with multiple parasites; thus, fully understanding interactions between individual parasite species and their hosts depends on accurate characterization of the parasite community. For parasitic nematodes, noninvasive methods for obtaining quantitative, species-specific infection data in wildlife are often unreliable. Consequently, characterization of gastrointestinal nematode communities of wild hosts has largely relied on lethal sampling to isolate and enumerate adult worms directly from the tissues of dead hosts. The necessity of lethal sampling severely restricts the host species that can be studied, the adequacy of sample sizes to assess diversity, the geographic scope of collections and the research questions that can be addressed. Focusing on gastrointestinal nematodes of wild African buffalo, we evaluated whether accurate characterization of nematode communities could be made using a noninvasive technique that combined conventional parasitological approaches with molecular barcoding. To establish the reliability of this new method, we compared estimates of gastrointestinal nematode abundance, prevalence, richness and community composition derived from lethal sampling with estimates derived from our noninvasive approach. Our noninvasive technique accurately estimated total and species-specific worm abundances, as well as worm prevalence and community composition when compared to the lethal sampling method. Importantly, the rate of parasite species discovery was similar for both methods, and only a modest number of barcoded larvae (n = 10) were needed to capture key aspects of parasite community composition. Overall, this new noninvasive strategy offers numerous advantages over lethal sampling methods for studying nematode-host interactions in wildlife and can readily be applied to a range of study systems. © 2015 John Wiley & Sons Ltd.

Novel methods of time-resolved fluorescence data analysis for in-vivo tissue characterization: application to atherosclerosis.

PubMed

Jo, J A; Fang, Q; Papaioannou, T; Qiao, J H; Fishbein, M C; Dorafshar, A; Reil, T; Baker, D; Freischlag, J; Marcu, L

2004-01-01

This study investigates the ability of new analytical methods of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) data to characterize tissue in-vivo, such as the composition of atherosclerotic vulnerable plaques. A total of 73 TR-LIFS measurements were taken in-vivo from the aorta of 8 rabbits, and subsequently analyzed using the Laguerre deconvolution technique. The investigated spots were classified as normal aorta, thin or thick lesions, and lesions rich in either collagen or macrophages/foam-cells. Different linear and nonlinear classification algorithms (linear discriminant analysis, stepwise linear discriminant analysis, principal component analysis, and feedforward neural networks) were developed using spectral and TR features (ratios of intensity values and Laguerre expansion coefficients, respectively). Normal intima and thin lesions were discriminated from thick lesions (sensitivity >90%, specificity 100%) using only spectral features. However, both spectral and time-resolved features were necessary to discriminate thick lesions rich in collagen from thick lesions rich in foam cells (sensitivity >85%, specificity >93%), and thin lesions rich in foam cells from normal aorta and thin lesions rich in collagen (sensitivity >85%, specificity >94%). Based on these findings, we believe that TR-LIFS information derived from the Laguerre expansion coefficients can provide a valuable additional dimension for in-vivo tissue characterization.

Cleavage of nucleic acids

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor L.; Brow, Mary Ann D.; Dahlberg, James E.

2007-12-11

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Invasive cleavage of nucleic acids

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann D.; Dahlberg, James E.

1999-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Invasive cleavage of nucleic acids

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann D.; Dahlberg, James E.

2002-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Cleavage of nucleic acids

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow; Mary Ann D.; Dahlberg, James E.

2010-11-09

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Cleavage of nucleic acids

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann D.; Dahlberg, James E.

2000-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Nucleic acid detection assays

DOEpatents

Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann; Dahlberg, James E.

2005-04-05

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Functional characterization of mouse spinal cord infiltrating CD8+ lymphocytes

PubMed Central

Deb, Chandra; Howe, Charles L

2011-01-01

Understanding the immunopathogenesis of neuroimmunological diseases of the CNS requires a robust method for isolating and characterizing the immune effector cells that infiltrate the spinal cord in animal models. We have developed a simple and rapid isolation method that produces high yields of spinal cord infiltrating leukocytes from a single demyelinated spinal cord and which maintains high surface expression of key immunophenotyping antigens. Using this method and the Theiler’s virus model of chronic demyelination, we report the presence of spinal cord infiltrating acute effector CD8+ lymphocytes that are CD45hiCD44loCD62L− and a population of spinal cord infiltrating target effector memory CD8+ lymphocytes that are CD45hiCD44hiCD62L−. These cells respond robustly to ex vivo stimulation by producing interferon γ but do not exhibit specificity for Theiler’s virus in a cytotoxicity assay. We conclude that target-derived lymphocytes in a mouse model of chronic spinal cord demyelination may have unique functional specificities. PMID:19596449

A look-up table based approach to characterize crystal twinning for synchrotron X-ray Laue microdiffraction scans

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

Li, Yao; Wan, Liang; Chen, Kai

An automated method has been developed to characterize the type and spatial distribution of twinning in crystal orientation maps from synchrotron X-ray Laue microdiffraction results. The method relies on a look-up table approach. Taking into account the twin axis and twin plane for plausible rotation and reflection twins, respectively, and the point group symmetry operations for a specific crystal, a look-up table listing crystal-specific rotation angle–axis pairs, which reveal the orientation relationship between the twin and the parent lattice, is generated. By comparing these theoretical twin–parent orientation relationships in the look-up table with the measured misorientations, twin boundaries are mappedmore » automatically from Laue microdiffraction raster scans with thousands of data points. Finally, taking advantage of the high orientation resolution of the Laue microdiffraction method, this automated approach is also applicable to differentiating twinning elements among multiple twinning modes in any crystal system.« less

A look-up table based approach to characterize crystal twinning for synchrotron X-ray Laue microdiffraction scans

DOE PAGES

Li, Yao; Wan, Liang; Chen, Kai

2015-04-25

An automated method has been developed to characterize the type and spatial distribution of twinning in crystal orientation maps from synchrotron X-ray Laue microdiffraction results. The method relies on a look-up table approach. Taking into account the twin axis and twin plane for plausible rotation and reflection twins, respectively, and the point group symmetry operations for a specific crystal, a look-up table listing crystal-specific rotation angle–axis pairs, which reveal the orientation relationship between the twin and the parent lattice, is generated. By comparing these theoretical twin–parent orientation relationships in the look-up table with the measured misorientations, twin boundaries are mappedmore » automatically from Laue microdiffraction raster scans with thousands of data points. Finally, taking advantage of the high orientation resolution of the Laue microdiffraction method, this automated approach is also applicable to differentiating twinning elements among multiple twinning modes in any crystal system.« less

Photogrammetry and Videogrammetry Methods Development for Solar Sail Structures. Masters Thesis awarded by George Washington Univ.

NASA Technical Reports Server (NTRS)

Pappa, Richard S. (Technical Monitor); Black, Jonathan T.

2003-01-01

This report discusses the development and application of metrology methods called photogrammetry and videogrammetry that make accurate measurements from photographs. These methods have been adapted for the static and dynamic characterization of gossamer structures, as four specific solar sail applications demonstrate. The applications prove that high-resolution, full-field, non-contact static measurements of solar sails using dot projection photogrammetry are possible as well as full-field, non-contact, dynamic characterization using dot projection videogrammetry. The accuracy of the measurement of the resonant frequencies and operating deflection shapes that were extracted surpassed expectations. While other non-contact measurement methods exist, they are not full-field and require significantly more time to take data.

Characterization of Cerebral White Matter Properties Using Quantitative Magnetic Resonance Imaging Stains

PubMed Central

Hurley, Samuel A.; Samsonov, Alexey A.; Adluru, Nagesh; Hosseinbor, Ameer Pasha; Mossahebi, Pouria; Tromp, Do P.M.; Zakszewski, Elizabeth; Field, Aaron S.

2011-01-01

Abstract The image contrast in magnetic resonance imaging (MRI) is highly sensitive to several mechanisms that are modulated by the properties of the tissue environment. The degree and type of contrast weighting may be viewed as image filters that accentuate specific tissue properties. Maps of quantitative measures of these mechanisms, akin to microstructural/environmental-specific tissue stains, may be generated to characterize the MRI and physiological properties of biological tissues. In this article, three quantitative MRI (qMRI) methods for characterizing white matter (WM) microstructural properties are reviewed. All of these measures measure complementary aspects of how water interacts with the tissue environment. Diffusion MRI, including diffusion tensor imaging, characterizes the diffusion of water in the tissues and is sensitive to the microstructural density, spacing, and orientational organization of tissue membranes, including myelin. Magnetization transfer imaging characterizes the amount and degree of magnetization exchange between free water and macromolecules like proteins found in the myelin bilayers. Relaxometry measures the MRI relaxation constants T1 and T2, which in WM have a component associated with the water trapped in the myelin bilayers. The conduction of signals between distant brain regions occurs primarily through myelinated WM tracts; thus, these methods are potential indicators of pathology and structural connectivity in the brain. This article provides an overview of the qMRI stain mechanisms, acquisition and analysis strategies, and applications for these qMRI stains. PMID:22432902

Detection of nucleic acid sequences by invader-directed cleavage

DOEpatents

Brow, Mary Ann D.; Hall, Jeff Steven Grotelueschen; Lyamichev, Victor; Olive, David Michael; Prudent, James Robert

1999-01-01

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The 5' nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based by charge.

A DC electrophoresis method for determining electrophoretic mobility through the pressure driven negation of electro osmosis

NASA Astrophysics Data System (ADS)

Karam, Pascal; Pennathur, Sumita

2016-11-01

Characterization of the electrophoretic mobility and zeta potential of micro and nanoparticles is important for assessing properties such as stability, charge and size. In electrophoretic techniques for such characterization, the bulk fluid motion due to the interaction between the fluid and the charged surface must be accounted for. Unlike current industrial systems which rely on DLS and oscillating potentials to mitigate electroosmotic flow (EOF), we propose a simple alternative electrophoretic method for optically determining electrophoretic mobility using a DC electric fields. Specifically, we create a system where an adverse pressure gradient counters EOF, and design the geometry of the channel so that the flow profile of the pressure driven flow matches that of the EOF in large regions of the channel (ie. where we observe particle flow). Our specific COMSOL-optimized geometry is two large cross sectional areas adjacent to a central, high aspect ratio channel. We show that this effectively removes EOF from a large region of the channel and allows for the accurate optical characterization of electrophoretic particle mobility, no matter the wall charge or particle size.

Principle, system, and applications of tip-enhanced Raman spectroscopy

NASA Astrophysics Data System (ADS)

Zhang, MingQian; Wang, Rui; Wu, XiaoBin; Wang, Jia

2012-08-01

Raman spectroscopy is a powerful technique in chemical information characterization. However, this spectral method is subject to two obstacles in nano-material detection. One is diffraction limited spatial resolution, and the other is its inherent small Raman cross section and weak signaling. To resolve these problems, a new approach has been developed, denoted as tip-enhanced Raman spectroscopy (TERS). TERS is capable of high-resolution and high-sensitivity detection and demonstrated to be a promising spectroscopic and micro-topographic method to characterize nano-materials and nanostructures. In this paper, the principle and experimental system of TERS are discussed. The latest application of TERS in molecule detection, biological specimen identification, nanao-material characterization, and semi-conductor material determination with some specific experimental examples are presented.

Statistical methods for analysis of radiation effects with tumor and dose location-specific information with application to the WECARE study of asynchronous contralateral breast cancer

PubMed Central

Langholz, Bryan; Thomas, Duncan C.; Stovall, Marilyn; Smith, Susan A.; Boice, John D.; Shore, Roy E.; Bernstein, Leslie; Lynch, Charles F.; Zhang, Xinbo; Bernstein, Jonine L.

2009-01-01

Summary Methods for the analysis of individually matched case-control studies with location-specific radiation dose and tumor location information are described. These include likelihood methods for analyses that just use cases with precise location of tumor information and methods that also include cases with imprecise tumor location information. The theory establishes that each of these likelihood based methods estimates the same radiation rate ratio parameters, within the context of the appropriate model for location and subject level covariate effects. The underlying assumptions are characterized and the potential strengths and limitations of each method are described. The methods are illustrated and compared using the WECARE study of radiation and asynchronous contralateral breast cancer. PMID:18647297

Insights from native mass spectrometry approaches for top- and middle- level characterization of site-specific antibody-drug conjugates

PubMed Central

Botzanowski, Thomas; Erb, Stéphane; Hernandez-Alba, Oscar; Ehkirch, Anthony; Colas, Olivier; Wagner-Rousset, Elsa; Rabuka, David; Beck, Alain; Drake, Penelope M.; Cianférani, Sarah

2017-01-01

ABSTRACT Antibody-drug conjugates (ADCs) have emerged as a family of compounds with promise as efficient immunotherapies. First-generation ADCs were generated mostly via reactions on either lysine side-chain amines or cysteine thiol groups after reduction of the interchain disulfide bonds, resulting in heterogeneous populations with a variable number of drug loads per antibody. To control the position and the number of drug loads, new conjugation strategies aiming at the generation of more homogeneous site-specific conjugates have been developed. We report here the first multi-level characterization of a site-specific ADC by state-of-the-art mass spectrometry (MS) methods, including native MS and its hyphenation to ion mobility (IM-MS). We demonstrate the versatility of native MS methodologies for site-specific ADC analysis, with the unique ability to provide several critical quality attributes within one single run, along with a direct snapshot of ADC homogeneity/heterogeneity without extensive data interpretation. The capabilities of native IM-MS to directly access site-specific ADC conformational information are also highlighted. Finally, the potential of these techniques for assessing an ADC's heterogeneity/homogeneity is illustrated by comparing the analytical characterization of a site-specific DAR4 ADC to that of first-generation ADCs. Altogether, our results highlight the compatibility, versatility, and benefits of native MS approaches for the analytical characterization of all types of ADCs, including site-specific conjugates. Thus, we envision integrating native MS and IM-MS approaches, even in their latest state-of-the-art forms, into workflows that benchmark bioconjugation strategies. PMID:28406343

Detection of Memory B Activity Against a Therapeutic Protein in Treatment-Naïve Subjects.

PubMed

Liao, Karen; Derbyshire, Stacy; Wang, Kai-Fen; Caucci, Cherilyn; Tang, Shuo; Holland, Claire; Loercher, Amy; Gunn, George R

2018-03-16

Bridging immunoassays commonly used to detect and characterize immunogenicity during biologic development do not provide direct information on the presence or development of a memory anti-drug antibody (ADA) response. In this study, a B cell ELISPOT assay method was used to evaluate pre-existing ADA for anti-TNFR1 domain antibody, GSK1995057, an experimental biologic in treatment naive subjects. This assay utilized a 7-day activation of PBMCs by a combination of GSK1995057 (antigen) and polyclonal stimulator followed by GSK1995057-specific ELISPOT for the enumeration of memory B cells that have differentiated into antibody secreting cells (ASC) in vitro. We demonstrated that GSK1995057-specific ASC were detectable in treatment-naïve subjects with pre-existing ADA; the frequency of drug-specific ASC was low and ranged from 1 to 10 spot forming units (SFU) per million cells. Interestingly, the frequency of drug-specific ASC correlated with the ADA level measured using an in vitro ADA assay. We further confirmed that the ASC originated from CD27 + memory B cells, not from CD27 - -naïve B cells. Our data demonstrated the utility of the B cell ELISPOT method in therapeutic protein immunogenicity evaluation, providing a novel way to confirm and characterize the cell population producing pre-existing ADA. This novel application of a B cell ELISPOT assay informs and characterizes immune memory activity regarding incidence and magnitude associated with a pre-existing ADA response.

Evaluation of field methods for vertical high resolution aquifer characterization

NASA Astrophysics Data System (ADS)

Vienken, T.; Tinter, M.; Rogiers, B.; Leven, C.; Dietrich, P.

2012-12-01

The delineation and characterization of subsurface (hydro)-stratigraphic structures is one of the challenging tasks of hydrogeological site investigations. The knowledge about the spatial distribution of soil specific properties and hydraulic conductivity (K) is the prerequisite for understanding flow and fluid transport processes. This is especially true for heterogeneous unconsolidated sedimentary deposits with a complex sedimentary architecture. One commonly used approach to investigate and characterize sediment heterogeneity is soil sampling and lab analyses, e.g. grain size distribution. Tests conducted on 108 samples show that calculation of K based on grain size distribution is not suitable for high resolution aquifer characterization of highly heterogeneous sediments due to sampling effects and large differences of calculated K values between applied formulas (Vienken & Dietrich 2011). Therefore, extensive tests were conducted at two test sites under different geological conditions to evaluate the performance of innovative Direct Push (DP) based approaches for the vertical high resolution determination of K. Different DP based sensor probes for the in-situ subsurface characterization based on electrical, hydraulic, and textural soil properties were used to obtain high resolution vertical profiles. The applied DP based tools proved to be a suitable and efficient alternative to traditional approaches. Despite resolution differences, all of the applied methods captured the main aquifer structure. Correlation of the DP based K estimates and proxies with DP based slug tests show that it is possible to describe the aquifer hydraulic structure on less than a meter scale by combining DP slug test data and continuous DP measurements. Even though correlations are site specific and appropriate DP tools must be chosen, DP is reliable and efficient alternative for characterizing even strongly heterogeneous sites with complex structured sedimentary aquifers (Vienken et al. 2012). References: Vienken, T., Leven, C., and Dietrich, P. 2012. Use of CPT and other direct push methods for (hydro-) stratigraphic aquifer characterization — a field study. Canadian Geotechnical Journal, 49(2): 197-206. Vienken, T., and Dietrich, P. 2011. Field evaluation of methods for determining hydraulic conductivity from grain size data. Journal of Hydrology, 400(1-2): 58-71.

A Method for Improved Interpretation of "Spot" Biomarker Data ...

EPA Pesticide Factsheets

A Method for Improved Interpretation of "Spot" Biomarker Data The National Exposure Research Laboratory (NERL) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA mission to protect human health and the environment. HEASD research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.

The Item-Specific Deficit Approach to evaluating verbal memory dysfunction: rationale, psychometrics, and application.

PubMed

Wright, Matthew J; Woo, Ellen; Schmitter-Edgecombe, Maureen; Hinkin, Charles H; Miller, Eric N; Gooding, Amanda L

2009-10-01

In the current study, we introduce the Item-Specific Deficit Approach (ISDA), a novel method for characterizing memory process deficits in list-learning data. To meet this objective, we applied the ISDA to California Verbal Learning Test (CVLT) data collected from a sample of 132 participants (53 healthy participants and 79 neurologically compromised participants). Overall, the ISDA indices measuring encoding, consolidation, and retrieval deficits demonstrated advantages over some traditional indices and indicated acceptable reliability and validity. Currently, the ISDA is intended for experimental use, although further research may support its utility for characterizing memory impairments in clinical assessments.

The integrated contaminant elution and tracer test toolkit, ICET3, for improved characterization of mass transfer, attenuation, and mass removal

NASA Astrophysics Data System (ADS)

Brusseau, Mark L.; Guo, Zhilin

2018-01-01

It is evident based on historical data that groundwater contaminant plumes persist at many sites, requiring costly long-term management. High-resolution site-characterization methods are needed to support accurate risk assessments and to select, design, and operate effective remediation operations. Most subsurface characterization methods are generally limited in their ability to provide unambiguous, real-time delineation of specific processes affecting mass-transfer, transformation, and mass removal, and accurate estimation of associated rates. An integrated contaminant elution and tracer test toolkit, comprising a set of local-scale groundwater extraction-and injection tests, was developed to ameliorate the primary limitations associated with standard characterization methods. The test employs extended groundwater extraction to stress the system and induce hydraulic and concentration gradients. Clean water can be injected, which removes the resident aqueous contaminant mass present in the higher-permeability zones and isolates the test zone from the surrounding plume. This ensures that the concentrations and fluxes measured within the isolated area are directly and predominantly influenced by the local mass-transfer and transformation processes controlling mass removal. A suite of standard and novel tracers can be used to delineate specific mass-transfer and attenuation processes that are active at a given site, and to quantify the associated mass-transfer and transformation rates. The conceptual basis for the test is first presented, followed by an illustrative application based on simulations produced with a 3-D mathematical model and a brief case study application.

Numerical Characterization of Piezoceramics Using Resonance Curves

PubMed Central

Pérez, Nicolás; Buiochi, Flávio; Brizzotti Andrade, Marco Aurélio; Adamowski, Julio Cezar

2016-01-01

Piezoelectric materials characterization is a challenging problem involving physical concepts, electrical and mechanical measurements and numerical optimization techniques. Piezoelectric ceramics such as Lead Zirconate Titanate (PZT) belong to the 6 mm symmetry class, which requires five elastic, three piezoelectric and two dielectric constants to fully represent the material properties. If losses are considered, the material properties can be represented by complex numbers. In this case, 20 independent material constants are required to obtain the full model. Several numerical methods have been used to adjust the theoretical models to the experimental results. The continuous improvement of the computer processing ability has allowed the use of a specific numerical method, the Finite Element Method (FEM), to iteratively solve the problem of finding the piezoelectric constants. This review presents the recent advances in the numerical characterization of 6 mm piezoelectric materials from experimental electrical impedance curves. The basic strategy consists in measuring the electrical impedance curve of a piezoelectric disk, and then combining the Finite Element Method with an iterative algorithm to find a set of material properties that minimizes the difference between the numerical impedance curve and the experimental one. Different methods to validate the results are also discussed. Examples of characterization of some common piezoelectric ceramics are presented to show the practical application of the described methods. PMID:28787875

Numerical Characterization of Piezoceramics Using Resonance Curves.

PubMed

Pérez, Nicolás; Buiochi, Flávio; Brizzotti Andrade, Marco Aurélio; Adamowski, Julio Cezar

2016-01-27

Piezoelectric materials characterization is a challenging problem involving physical concepts, electrical and mechanical measurements and numerical optimization techniques. Piezoelectric ceramics such as Lead Zirconate Titanate (PZT) belong to the 6 mm symmetry class, which requires five elastic, three piezoelectric and two dielectric constants to fully represent the material properties. If losses are considered, the material properties can be represented by complex numbers. In this case, 20 independent material constants are required to obtain the full model. Several numerical methods have been used to adjust the theoretical models to the experimental results. The continuous improvement of the computer processing ability has allowed the use of a specific numerical method, the Finite Element Method (FEM), to iteratively solve the problem of finding the piezoelectric constants. This review presents the recent advances in the numerical characterization of 6 mm piezoelectric materials from experimental electrical impedance curves. The basic strategy consists in measuring the electrical impedance curve of a piezoelectric disk, and then combining the Finite Element Method with an iterative algorithm to find a set of material properties that minimizes the difference between the numerical impedance curve and the experimental one. Different methods to validate the results are also discussed. Examples of characterization of some common piezoelectric ceramics are presented to show the practical application of the described methods.

Filtration Characterization Method as Tool to Assess Membrane Bioreactor Sludge Filterability—The Delft Experience

PubMed Central

Lousada-Ferreira, Maria; Krzeminski, Pawel; Geilvoet, Stefan; Moreau, Adrien; Gil, Jose A.; Evenblij, Herman; van Lier, Jules B.; van der Graaf, Jaap H. J. M.

2014-01-01

Prevention and removal of fouling is often the most energy intensive process in Membrane Bioreactors (MBRs), responsible for 40% to 50% of the total specific energy consumed in submerged MBRs. In the past decade, methods were developed to quantify and qualify fouling, aiming to support optimization in MBR operation. Therefore, there is a need for an evaluation of the lessons learned and how to proceed. In this article, five different methods for measuring MBR activated sludge filterability and critical flux are described, commented and evaluated. Both parameters characterize the fouling potential in full-scale MBRs. The article focuses on the Delft Filtration Characterization method (DFCm) as a convenient tool to characterize sludge properties, namely on data processing, accuracy, reproducibility, reliability, and applicability, defining the boundaries of the DFCm. Significant progress was made concerning fouling measurements in particular by using straight forward approaches focusing on the applicability of the obtained results. Nevertheless, a fouling measurement method is still to be defined which is capable of being unequivocal, concerning the fouling parameters definitions; practical and simple, in terms of set-up and operation; broad and useful, in terms of obtained results. A step forward would be the standardization of the aforementioned method to assess the sludge filtration quality. PMID:24957174

Reflexion measurements for inverse characterization of steel diffusion bond mechanical properties

NASA Astrophysics Data System (ADS)

Le Bourdais, Florian; Cachon, Lionel; Rigal, Emmanuel

2017-02-01

The present work describes a non-destructive testing method aimed at securing high manufacturing quality of the innovative compact heat exchanger developed under the framework of the CEA R&D program dedicated to the Advanced Sodium Technological Reactor for Industrial Demonstration (ASTRID). The heat exchanger assembly procedure currently proposed involves high temperature and high pressure diffusion welding of stainless steel plates. The aim of the non-destructive method presented herein is to characterize the quality of the welds obtained through this assembly process. Based on a low-frequency model developed by Baik and Thompson [1], pulse-echo normal incidence measurements are calibrated according to a specific procedure and allow the determination of the welding interface stiffness using a nonlinear fitting procedure in the frequency domain. Performing the characterization of plates after diffusion welding using this method allows a useful assessment of the material state as a function of the diffusion bonding process.

Shallow Reflection Method for Water-Filled Void Detection and Characterization

NASA Astrophysics Data System (ADS)

Zahari, M. N. H.; Madun, A.; Dahlan, S. H.; Joret, A.; Hazreek, Z. A. M.; Mohammad, A. H.; Izzaty, R. A.

2018-04-01

Shallow investigation is crucial in enhancing the characteristics of subsurface void commonly encountered in civil engineering, and one such technique commonly used is seismic-reflection technique. An assessment of the effectiveness of such an approach is critical to determine whether the quality of the works meets the prescribed requirements. Conventional quality testing suffers limitations including: limited coverage (both area and depth) and problems with resolution quality. Traditionally quality assurance measurements use laboratory and in-situ invasive and destructive tests. However geophysical approaches, which are typically non-invasive and non-destructive, offer a method by which improvement of detection can be measured in a cost-effective way. Of this seismic reflection have proved useful to assess void characteristic, this paper evaluates the application of shallow seismic-reflection method in characterizing the water-filled void properties at 0.34 m depth, specifically for detection and characterization of void measurement using 2-dimensional tomography.

Characterization of coronary plaque regions in intravascular ultrasound images using a hybrid ensemble classifier.

PubMed

Hwang, Yoo Na; Lee, Ju Hwan; Kim, Ga Young; Shin, Eun Seok; Kim, Sung Min

2018-01-01

The purpose of this study was to propose a hybrid ensemble classifier to characterize coronary plaque regions in intravascular ultrasound (IVUS) images. Pixels were allocated to one of four tissues (fibrous tissue (FT), fibro-fatty tissue (FFT), necrotic core (NC), and dense calcium (DC)) through processes of border segmentation, feature extraction, feature selection, and classification. Grayscale IVUS images and their corresponding virtual histology images were acquired from 11 patients with known or suspected coronary artery disease using 20 MHz catheter. A total of 102 hybrid textural features including first order statistics (FOS), gray level co-occurrence matrix (GLCM), extended gray level run-length matrix (GLRLM), Laws, local binary pattern (LBP), intensity, and discrete wavelet features (DWF) were extracted from IVUS images. To select optimal feature sets, genetic algorithm was implemented. A hybrid ensemble classifier based on histogram and texture information was then used for plaque characterization in this study. The optimal feature set was used as input of this ensemble classifier. After tissue characterization, parameters including sensitivity, specificity, and accuracy were calculated to validate the proposed approach. A ten-fold cross validation approach was used to determine the statistical significance of the proposed method. Our experimental results showed that the proposed method had reliable performance for tissue characterization in IVUS images. The hybrid ensemble classification method outperformed other existing methods by achieving characterization accuracy of 81% for FFT and 75% for NC. In addition, this study showed that Laws features (SSV and SAV) were key indicators for coronary tissue characterization. The proposed method had high clinical applicability for image-based tissue characterization. Copyright © 2017 Elsevier B.V. All rights reserved.

Group Theoretical Characterization of Wave Equations

NASA Astrophysics Data System (ADS)

Nisticò, Giuseppe

2017-12-01

Group theoretical methods, worked out in particular by Mackey and Wigner, allow to attain the explicit Quantum Theory of a free particle through a purely deductive development based on symmetry principles. The extension of these methods to the case of an interacting particle finds a serious obstacle in the loss of the symmetry condition for the transformations of Galilei's group. The known attempts towards such an extension introduce restrictions which lead to theories empirically too limited. In the present article we show how the difficulties raised by the loss of symmetry can be overcome without the restrictions that affect tha past attempts. According to our results, the different specific forms of the wave equation of an interacting particle are implied by particular first order invariance properties that characterize the interaction with respect to specific sub-groups of galileian transformations. Moreover, the possibility of yet unknown forms of the wave equation is left open.

Molecular diagnostics for the detection and characterization of microbial pathogens.

PubMed

Procop, Gary W

2007-09-01

New and advanced methods of molecular diagnostics are changing the way we practice clinical microbiology, which affects the practice of medicine. Signal amplification and real-time nucleic acid amplification technologies offer a sensitive and specific result with a more rapid turnaround time than has ever before been possible. Numerous methods of postamplification analysis afford the simultaneous detection and differentiation of numerous microbial pathogens, their mechanisms of resistance, and the construction of disease-specific assays. The technical feasibility of these assays has already been demonstrated. How these new, often more expensive tests will be incorporated into routine practice and the impact they will have on patient care remain to be determined. One of the most attractive uses for such techniques is to achieve a more rapid characterization of the infectious agent so that a narrower-spectrum antimicrobial agent may be used, which should have an impact on resistance patterns.

A new analytical method for characterizing nonlinear visual processes with stimuli of arbitrary distribution: Theory and applications.

PubMed

Hayashi, Ryusuke; Watanabe, Osamu; Yokoyama, Hiroki; Nishida, Shin'ya

2017-06-01

Characterization of the functional relationship between sensory inputs and neuronal or observers' perceptual responses is one of the fundamental goals of systems neuroscience and psychophysics. Conventional methods, such as reverse correlation and spike-triggered data analyses are limited in their ability to resolve complex and inherently nonlinear neuronal/perceptual processes because these methods require input stimuli to be Gaussian with a zero mean. Recent studies have shown that analyses based on a generalized linear model (GLM) do not require such specific input characteristics and have advantages over conventional methods. GLM, however, relies on iterative optimization algorithms and its calculation costs become very expensive when estimating the nonlinear parameters of a large-scale system using large volumes of data. In this paper, we introduce a new analytical method for identifying a nonlinear system without relying on iterative calculations and yet also not requiring any specific stimulus distribution. We demonstrate the results of numerical simulations, showing that our noniterative method is as accurate as GLM in estimating nonlinear parameters in many cases and outperforms conventional, spike-triggered data analyses. As an example of the application of our method to actual psychophysical data, we investigated how different spatiotemporal frequency channels interact in assessments of motion direction. The nonlinear interaction estimated by our method was consistent with findings from previous vision studies and supports the validity of our method for nonlinear system identification.

Tools for the functional interpretation of metabolomic experiments.

PubMed

Chagoyen, Monica; Pazos, Florencio

2013-11-01

The so-called 'omics' approaches used in modern biology aim at massively characterizing the molecular repertories of living systems at different levels. Metabolomics is one of the last additions to the 'omics' family and it deals with the characterization of the set of metabolites in a given biological system. As metabolomic techniques become more massive and allow characterizing larger sets of metabolites, automatic methods for analyzing these sets in order to obtain meaningful biological information are required. Only recently the first tools specifically designed for this task in metabolomics appeared. They are based on approaches previously used in transcriptomics and other 'omics', such as annotation enrichment analysis. These, together with generic tools for metabolic analysis and visualization not specifically designed for metabolomics will for sure be in the toolbox of the researches doing metabolomic experiments in the near future.

Agricultural areas in potentially contaminated sites: characterization, risk, management.

PubMed

Vanni, Fabiana; Scaini, Federica; Beccaloni, Eleonora

2016-01-01

In Italy, the current legislation for contaminants in soils provides two land uses: residential/public or private gardens and commercial/industrial; there are not specific reference values for agricultural soils, even if a special decree has been developed and is currently going through the legislative approval process. The topic of agricultural areas is relevant, also in consideration of their presence near potentially contaminated sites. Aim and results. In this paper, contamination sources and transport modes of contaminants from sources to the target in agricultural areas are examined and a suitable "conceptual model" to define appropriate characterization methods and risk assessment procedures is proposed. These procedures have already been used by the National Institute of Health in various Italian areas characterized by different agricultural settings. Finally, specific remediation techniques are suggested to preserve soil resources and, if possible, its particular land use.

Allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), a powerful method for diagnosing loss of imprinting of the 11p15 region in Russell Silver and Beckwith Wiedemann syndromes.

PubMed

Azzi, Salah; Steunou, Virginie; Rousseau, Alexandra; Rossignol, Sylvie; Thibaud, Nathalie; Danton, Fabienne; Le Jule, Marilyne; Gicquel, Christine; Le Bouc, Yves; Netchine, Irène

2011-02-01

Many human syndromes involve a loss of imprinting (LOI) due to a loss (LOM) or a gain of DNA methylation (GOM). Most LOI occur as mosaics and can therefore be difficult to detect with conventional methods. The human imprinted 11p15 region is crucial for the control of fetal growth, and LOI at this locus is associated with two clinical disorders with opposite phenotypes: Beckwith-Wiedemann syndrome (BWS), characterized by fetal overgrowth and a high risk of tumors, and Russell-Silver syndrome (RSS), characterized by intrauterine and postnatal growth restriction. Until recently, we have been using Southern blotting for the diagnosis of RSS and BWS. We describe here a powerful quantitative technique, allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), for the diagnosis of these two complex disorders. We first checked the specificity of the probes and primers used for ASMM RTQ-PCR. We then carried out statistical validation for this method, on both retrospective and prospective populations of patients. This analysis demonstrated that ASMM RTQ-PCR is more sensitive than Southern blotting for detecting low degree of LOI. Moreover, ASMM RTQ-PCR is a very rapid, reliable, simple, safe, and cost effective method. © 2011 Wiley-Liss, Inc.

Quantitative impact characterization of aeronautical CFRP materials with non-destructive testing methods

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

Kiefel, Denis, E-mail: Denis.Kiefel@airbus.com, E-mail: Rainer.Stoessel@airbus.com; Stoessel, Rainer, E-mail: Denis.Kiefel@airbus.com, E-mail: Rainer.Stoessel@airbus.com; Grosse, Christian, E-mail: Grosse@tum.de

2015-03-31

In recent years, an increasing number of safety-relevant structures are designed and manufactured from carbon fiber reinforced polymers (CFRP) in order to reduce weight of airplanes by taking the advantage of their specific strength into account. Non-destructive testing (NDT) methods for quantitative defect analysis of damages are liquid- or air-coupled ultrasonic testing (UT), phased array ultrasonic techniques, and active thermography (IR). The advantage of these testing methods is the applicability on large areas. However, their quantitative information is often limited on impact localization and size. In addition to these techniques, Airbus Group Innovations operates a micro x-ray computed tomography (μ-XCT)more » system, which was developed for CFRP characterization. It is an open system which allows different kinds of acquisition, reconstruction, and data evaluation. One main advantage of this μ-XCT system is its high resolution with 3-dimensional analysis and visualization opportunities, which enables to gain important quantitative information for composite part design and stress analysis. Within this study, different NDT methods will be compared at CFRP samples with specified artificial impact damages. The results can be used to select the most suitable NDT-method for specific application cases. Furthermore, novel evaluation and visualization methods for impact analyzes are developed and will be presented.« less

Numerical analysis of Venous External Scaffolding Technology for Saphenous Vein Grafts.

PubMed

Meirson, T; Orion, E; Avrahami, I

2015-07-16

This paper presents a method for analyzing and comparing numerically Saphenous Vein Grafts (SVGs) following Coronary Artery Bypass Graft surgery (CABG). The method analyses the flow dynamics inside vein grafts with and without supporting using Venous External Scaffolding Technology (VEST). The numerical method uses patients׳ specific computational fluid dynamics (CFD) methods to characterize the relevant hemodynamic parameters of patients׳ SVGs. The method was used to compare the hemodynamics of six patient׳s specific model and flow conditions of stented and non-stented SVGs, 12 months post-transplantation. The flow parameters used to characterize the grafts׳ hemodynamics include Time Averaged Wall Shear Stress (TAWSS), Oscillatory Shear Index (OSI) and Relative Residence Time (RRT). The effect of stenting was clearly demonstrated by the chosen parameters. SVGs under constriction of VEST were associated with similar spatial average of TAWSS (10.73 vs 10.29 dyn/cm(2)), yet had fewer lesions with low TAWSS, lower OSI (0.041 vs 0.08) and RRT (0.12 vs 0.24), and more uniform flow with less flow discrepancies. In conclusion, the suggested method and parameters well demonstrated the advantage of VEST support. Stenting vein grafts with VEST improved hemodynamic factors which are correlated to graft failure following CABG procedure. Copyright © 2015 Elsevier Ltd. All rights reserved.

Color-coded automated signal intensity curves for detection and characterization of breast lesions: preliminary evaluation of a new software package for integrated magnetic resonance-based breast imaging.

PubMed

Pediconi, Federica; Catalano, Carlo; Venditti, Fiammetta; Ercolani, Mauro; Carotenuto, Luigi; Padula, Simona; Moriconi, Enrica; Roselli, Antonella; Giacomelli, Laura; Kirchin, Miles A; Passariello, Roberto

2005-07-01

The objective of this study was to evaluate the value of a color-coded automated signal intensity curve software package for contrast-enhanced magnetic resonance mammography (CE-MRM) in patients with suspected breast cancer. Thirty-six women with suspected breast cancer based on mammographic and sonographic examinations were preoperatively evaluated on CE-MRM. CE-MRM was performed on a 1.5-T magnet using a 2D Flash dynamic T1-weighted sequence. A dosage of 0.1 mmol/kg of Gd-BOPTA was administered at a flow rate of 2 mL/s followed by 10 mL of saline. Images were analyzed with the new software package and separately with a standard display method. Statistical comparison was performed of the confidence for lesion detection and characterization with the 2 methods and of the diagnostic accuracy for characterization compared with histopathologic findings. At pathology, 54 malignant lesions and 14 benign lesions were evaluated. All 68 (100%) lesions were detected with both methods and good correlation with histopathologic specimens was obtained. Confidence for both detection and characterization was significantly (P < or = 0.025) better with the color-coded method, although no difference (P > 0.05) between the methods was noted in terms of the sensitivity, specificity, and overall accuracy for lesion characterization. Excellent agreement between the 2 methods was noted for both the determination of lesion size (kappa = 0.77) and determination of SI/T curves (kappa = 0.85). The novel color-coded signal intensity curve software allows lesions to be visualized as false color maps that correspond to conventional signal intensity time curves. Detection and characterization of breast lesions with this method is quick and easily interpretable.

Molecular characterization and identification of members of the Anopheles subpictus complex in Sri Lanka

PubMed Central

2013-01-01

Background Anopheles subpictus sensu lato is a major malaria vector in South and Southeast Asia. Based initially on polytene chromosome inversion polymorphism, and subsequently on morphological characterization, four sibling species A-D were reported from India. The present study uses molecular methods to further characterize and identify sibling species in Sri Lanka. Methods Mosquitoes from Sri Lanka were morphologically identified to species and sequenced for the ribosomal internal transcribed spacer-2 (ITS2) and the mitochondrial cytochrome c oxidase subunit-I (COI) genes. These sequences, together with others from GenBank, were used to construct phylogenetic trees and parsimony haplotype networks and to test for genetic population structure. Results Both ITS2 and COI sequences revealed two divergent clades indicating that the Subpictus complex in Sri Lanka is composed of two genetically distinct species that correspond to species A and species B from India. Phylogenetic analysis showed that species A and species B do not form a monophyletic clade but instead share genetic similarity with Anopheles vagus and Anopheles sundaicus s.l., respectively. An allele specific identification method based on ITS2 variation was developed for the reliable identification of species A and B in Sri Lanka. Conclusion Further multidisciplinary studies are needed to establish the species status of all chromosomal forms in the Subpictus complex. This study emphasizes the difficulties in using morphological characters for species identification in An. subpictus s.l. in Sri Lanka and demonstrates the utility of an allele specific identification method that can be used to characterize the differential bio-ecological traits of species A and B in Sri Lanka. PMID:24001126

Waste Analysis Plan and Waste Characterization Survey, Barksdale AFB, Louisiana

DTIC Science & Technology

1991-03-01

review to assess if analysis is needed, any analyses that are to be provided by generators, and methods to be used to meet specific waste analysis ...sampling method , sampling frequency, parameters of analysis , SW 846 test methods , Department of Transportation (DOT) shipping name and hazard class...S.e.iceA w/Atchs 2. HQ SAC/DEV Ltr, 28 Sep 90 19 119 APPENDIX B Waste Analysis Plan Rationale 21 APPENDIX B 1. SAMPLING METHOD RATIONALE: Composite Liquid

Framework for Investigating Asbestos-Contaminated Superfund Sites

EPA Science Inventory

The document presents a recommended framework for investigating and characterizing the potential for human exposure and risk from asbestos contamination in outdoor soil and indoor dust at Superfund removal and remedial sites. The framework presents specific strategies and method...

Lidar-Based Rock-Fall Hazard Characterization of Cliffs

USGS Publications Warehouse

Collins, Brian D.; Greg M.Stock,

2017-01-01

Rock falls from cliffs and other steep slopes present numerous challenges for detailed geological characterization. In steep terrain, rock-fall source areas are both dangerous and difficult to access, severely limiting the ability to make detailed structural and volumetric measurements necessary for hazard assessment. Airborne and terrestrial lidar survey methods can provide high-resolution data needed for volumetric, structural, and deformation analyses of rock falls, potentially making these analyses straightforward and routine. However, specific methods to collect, process, and analyze lidar data of steep cliffs are needed to maximize analytical accuracy and efficiency. This paper presents observations showing how lidar data sets should be collected, filtered, registered, and georeferenced to tailor their use in rock fall characterization. Additional observations concerning surface model construction, volumetric calculations, and deformation analysis are also provided.

In vitro uptake of apoptotic body mimicking phosphatidylserine-quantum dot micelles by monocytic cell line

NASA Astrophysics Data System (ADS)

Maiseyeu, Andrei; Bagalkot, Vaishali

2014-04-01

A new quantum dot (QD) PEGylated micelle laced with phosphatidylserine (PS) for specific scavenger receptor-mediated uptake by macrophages is reported. The size and surface chemistry of PS-QD micelles were characterized by standard methods and the effects of their physicochemical properties on specific targeting and uptake were comprehensively studied in a monocytic cell line (J774A.1).

Search for Patterns of Functional Specificity in the Brain: A Nonparametric Hierarchical Bayesian Model for Group fMRI Data

PubMed Central

Sridharan, Ramesh; Vul, Edward; Hsieh, Po-Jang; Kanwisher, Nancy; Golland, Polina

2012-01-01

Functional MRI studies have uncovered a number of brain areas that demonstrate highly specific functional patterns. In the case of visual object recognition, small, focal regions have been characterized with selectivity for visual categories such as human faces. In this paper, we develop an algorithm that automatically learns patterns of functional specificity from fMRI data in a group of subjects. The method does not require spatial alignment of functional images from different subjects. The algorithm is based on a generative model that comprises two main layers. At the lower level, we express the functional brain response to each stimulus as a binary activation variable. At the next level, we define a prior over sets of activation variables in all subjects. We use a Hierarchical Dirichlet Process as the prior in order to learn the patterns of functional specificity shared across the group, which we call functional systems, and estimate the number of these systems. Inference based on our model enables automatic discovery and characterization of dominant and consistent functional systems. We apply the method to data from a visual fMRI study comprised of 69 distinct stimulus images. The discovered system activation profiles correspond to selectivity for a number of image categories such as faces, bodies, and scenes. Among systems found by our method, we identify new areas that are deactivated by face stimuli. In empirical comparisons with perviously proposed exploratory methods, our results appear superior in capturing the structure in the space of visual categories of stimuli. PMID:21884803

Improved Phase Characterization of Far-Regional Body Wave Arrivals in Central Asia

DTIC Science & Technology

2008-09-30

developing array -based methods that can more accurately characterize far-regional (14*-29*) seismic wavefield structure. Far- regional (14*-29*) seismograms...arrivals with the primary arrivals. These complexities can be region and earthquake specific. The regional seismic arrays that have been built in the last...fifteen years should be a rich data source for the study of far-regional phase behavior. The arrays are composed of high-quality borehole seismometers

Characterization of spacecraft humidity condensate

NASA Technical Reports Server (NTRS)

Muckle, Susan; Schultz, John R.; Sauer, Richard L.

1994-01-01

When construction of Space Station Freedom reaches the Permanent Manned Capability (PMC) stage, the Water Recovery and Management Subsystem will be fully operational such that (distilled) urine, spent hygiene water, and humidity condensate will be reclaimed to provide water of potable quality. The reclamation technologies currently baselined to process these waste waters include adsorption, ion exchange, catalytic oxidation, and disinfection. To ensure that the baseline technologies will be able to effectively remove those compounds presenting a health risk to the crew, the National Research Council has recommended that additional information be gathered on specific contaminants in waste waters representative of those to be encountered on the Space Station. With the application of new analytical methods and the analysis of waste water samples more representative of the Space Station environment, advances in the identification of the specific contaminants continue to be made. Efforts by the Water and Food Analytical Laboratory at JSC were successful in enlarging the database of contaminants in humidity condensate. These efforts have not only included the chemical characterization of condensate generated during ground-based studies, but most significantly the characterization of cabin and Spacelab condensate generated during Shuttle missions. The analytical results presented in this paper will be used to show how the composition of condensate varies amongst enclosed environments and thus the importance of collecting condensate from an environment close to that of the proposed Space Station. Although advances were made in the characterization of space condensate, complete characterization, particularly of the organics, requires further development of analytical methods.

Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections.

PubMed

Verweij, Jaco J; Stensvold, C Rune

2014-04-01

Over the past few decades, nucleic acid-based methods have been developed for the diagnosis of intestinal parasitic infections. Advantages of nucleic acid-based methods are numerous; typically, these include increased sensitivity and specificity and simpler standardization of diagnostic procedures. DNA samples can also be stored and used for genetic characterization and molecular typing, providing a valuable tool for surveys and surveillance studies. A variety of technologies have been applied, and some specific and general pitfalls and limitations have been identified. This review provides an overview of the multitude of methods that have been reported for the detection of intestinal parasites and offers some guidance in applying these methods in the clinical laboratory and in epidemiological studies.

Molecular Testing for Clinical Diagnosis and Epidemiological Investigations of Intestinal Parasitic Infections

PubMed Central

Stensvold, C. Rune

2014-01-01

SUMMARY Over the past few decades, nucleic acid-based methods have been developed for the diagnosis of intestinal parasitic infections. Advantages of nucleic acid-based methods are numerous; typically, these include increased sensitivity and specificity and simpler standardization of diagnostic procedures. DNA samples can also be stored and used for genetic characterization and molecular typing, providing a valuable tool for surveys and surveillance studies. A variety of technologies have been applied, and some specific and general pitfalls and limitations have been identified. This review provides an overview of the multitude of methods that have been reported for the detection of intestinal parasites and offers some guidance in applying these methods in the clinical laboratory and in epidemiological studies. PMID:24696439

An ELISA DYRK1A non-radioactive kinase assay suitable for the characterization of inhibitors

PubMed Central

Liu, Yong; Adayev, Tatyana; Hwang, Yu-Wen

2017-01-01

The DYRK1A (dual specificity tyrosine phosphorylation-regulated kinase 1A) gene encodes a proline-directed Ser/Thr kinase. Elevated expression and/or altered distribution of the kinase have been implicated in the neurological impairments associated with Down syndrome (DS) and Alzheimer’s disease (AD). Consequently, DYRK1A inhibition has been of significant interest as a potential strategy for therapeutic intervention of DS and AD. Many classes of novel inhibitors have been described in the past decade. Although non-radioactive methods for analyzing DYRK1A inhibition have been developed, methods employing radioactive tracers are still commonly used for quantitative characterization of DYRK1A inhibitors. Here, we present a non-radioactive ELISA assay based on the detection of DYRK1A-phosphorylated dynamin 1a fragment using a phosphorylation site-specific antibody. The assay was verified by the use of two well-characterized DYRK1A inhibitors, epigallocatechin gallate (EGCG) and harmine. The IC 50s for EGCG and harmine determined by the ELISA method were found to be comparable to those previously measured by radioactive tracing methods.  Furthermore, we determined the mode of inhibition for EGCG and harmine by a modification of the ELISA assay. This assay confirms the mode of inhibition of EGCG (non-ATP-competitive) and harmine (ATP-competitive), as previously determined. We conclude that the ELISA platform demonstrated here is a viable alternative to the traditional radioactive tracer assays for analyzing DYRK1A inhibitors. PMID:28163906

Automatic 3D segmentation of multiphoton images: a key step for the quantification of human skin.

PubMed

Decencière, Etienne; Tancrède-Bohin, Emmanuelle; Dokládal, Petr; Koudoro, Serge; Pena, Ana-Maria; Baldeweck, Thérèse

2013-05-01

Multiphoton microscopy has emerged in the past decade as a useful noninvasive imaging technique for in vivo human skin characterization. However, it has not been used until now in evaluation clinical trials, mainly because of the lack of specific image processing tools that would allow the investigator to extract pertinent quantitative three-dimensional (3D) information from the different skin components. We propose a 3D automatic segmentation method of multiphoton images which is a key step for epidermis and dermis quantification. This method, based on the morphological watershed and graph cuts algorithms, takes into account the real shape of the skin surface and of the dermal-epidermal junction, and allows separating in 3D the epidermis and the superficial dermis. The automatic segmentation method and the associated quantitative measurements have been developed and validated on a clinical database designed for aging characterization. The segmentation achieves its goals for epidermis-dermis separation and allows quantitative measurements inside the different skin compartments with sufficient relevance. This study shows that multiphoton microscopy associated with specific image processing tools provides access to new quantitative measurements on the various skin components. The proposed 3D automatic segmentation method will contribute to build a powerful tool for characterizing human skin condition. To our knowledge, this is the first 3D approach to the segmentation and quantification of these original images. © 2013 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

Evaluation of crack status in a meter-size concrete structure using the ultrasonic nonlinear coda wave interferometry.

PubMed

Legland, Jean-Baptiste; Zhang, Yuxiang; Abraham, Odile; Durand, Olivier; Tournat, Vincent

2017-10-01

The field of civil engineering is in need of new methods of non-destructive testing, especially in order to prevent and monitor the serious deterioration of concrete structures. In this work, experimental results are reported on fault detection and characterization in a meter-scale concrete structure using an ultrasonic nonlinear coda wave interferometry (NCWI) method. This method entails the nonlinear mixing of strong pump waves with multiple scattered probe (coda) waves, along with analysis of the net effect using coda wave interferometry. A controlled damage protocol is implemented on a post-tensioned, meter-scale concrete structure in order to generate cracking within a specific area being monitored by NCWI. The nonlinear acoustic response due to the high amplitude of acoustic modulation yields information on the elastic nonlinearities of concrete, as evaluated by two specific nonlinear observables. The increase in nonlinearity level corresponds to the creation of a crack with a network of microcracks localized at its base. In addition, once the crack closes as a result of post-tensioning, the residual nonlinearities confirm the presence of the closed crack. Last, the benefits and applicability of this NCWI method to the characterization and monitoring of large structures are discussed.

A novel sample preparation and on-line HPLC-DAD-MS/MS-BCD analysis for rapid screening and characterization of specific enzyme inhibitors in herbal extracts: case study of α-glucosidase.

PubMed

Li, D Q; Zhao, J; Xie, J; Li, S P

2014-01-01

Drug discovery from complex mixture like Chinese herbs is a challenge and extensive false positives make the obtainment of specific bioactive compounds difficult. In the present study, a novel sample preparation method was proposed to rapidly reveal the specific bioactive compounds from complex mixtures using α-glucosidase as a case. Firstly, aqueous and methanol extracts of 500 traditional Chinese medicines were carried out with the aim of finding new sources of α-glucosidase inhibitors. As a result, the extracts of fruit of Terminalia chebula (FTC), flowers of Rosa rugosa (FRR) and Eugenia caryophyllata (FEC) as well as husk of Punica granatum (HPG) showed high inhibition on α-glucosidase. On-line liquid chromatography-diode array detection-tandem mass spectrometry and biochemical detection (HPLC-DAD-MS/MS-BCD) was performed to rapidly screen and characterize α-glucosidase inhibitors in these four extracts. After tentative identification, most of compounds with inhibitory activity in the investigated crude extracts were found to be tannins commonly recognized as non-specific enzyme inhibitors in vitro. Subsequently, the four extracts were treated with gelatin to improve specificity of the on-line system. Finally, two compounds with specific α-glucosidase inhibition were identified as corilagin and ellagic acid. The developed method could discover specific α-glucosidase inhibitors in complex mixtures such as plant extracts, which could also be used for discovery of specific inhibitors of other enzymes. Copyright © 2013 Elsevier B.V. All rights reserved.

Rapid Characterization of Microorganisms by Mass Spectrometry—What Can Be Learned and How?

NASA Astrophysics Data System (ADS)

Fenselau, Catherine C.

2013-08-01

Strategies for the rapid and reliable analysis of microorganisms have been sought to meet national needs in defense, homeland security, space exploration, food and water safety, and clinical diagnosis. Mass spectrometry has long been a candidate technique because it is extremely rapid and can provide highly specific information. It has excellent sensitivity. Molecular and fragment ion masses provide detailed fingerprints, which can also be interpreted. Mass spectrometry is also a broad band method—everything has a mass—and it is automatable. Mass spectrometry is a physiochemical method that is orthogonal and complementary to biochemical and morphological methods used to characterize microorganisms.

Molecular characterization and identification of members of the Anopheles subpictus complex in Sri Lanka.

PubMed

Surendran, Sinnathamby N; Sarma, Devojit K; Jude, Pavilupillai J; Kemppainen, Petri; Kanthakumaran, Nadarajah; Gajapathy, Kanapathy; Peiris, Lalanthika B S; Ramasamy, Ranjan; Walton, Catherine

2013-08-30

Anopheles subpictus sensu lato is a major malaria vector in South and Southeast Asia. Based initially on polytene chromosome inversion polymorphism, and subsequently on morphological characterization, four sibling species A-D were reported from India. The present study uses molecular methods to further characterize and identify sibling species in Sri Lanka. Mosquitoes from Sri Lanka were morphologically identified to species and sequenced for the ribosomal internal transcribed spacer-2 (ITS2) and the mitochondrial cytochrome c oxidase subunit-I (COI) genes. These sequences, together with others from GenBank, were used to construct phylogenetic trees and parsimony haplotype networks and to test for genetic population structure. Both ITS2 and COI sequences revealed two divergent clades indicating that the Subpictus complex in Sri Lanka is composed of two genetically distinct species that correspond to species A and species B from India. Phylogenetic analysis showed that species A and species B do not form a monophyletic clade but instead share genetic similarity with Anopheles vagus and Anopheles sundaicus s.l., respectively. An allele specific identification method based on ITS2 variation was developed for the reliable identification of species A and B in Sri Lanka. Further multidisciplinary studies are needed to establish the species status of all chromosomal forms in the Subpictus complex. This study emphasizes the difficulties in using morphological characters for species identification in An. subpictus s.l. in Sri Lanka and demonstrates the utility of an allele specific identification method that can be used to characterize the differential bio-ecological traits of species A and B in Sri Lanka.

Chemical Fingerprinting of Materials Developed Due to Environmental Issues

NASA Technical Reports Server (NTRS)

Smith, Doris A.; McCool, A. (Technical Monitor)

2000-01-01

Instrumental chemical analysis methods are developed and used to chemically fingerprint new and modified External Tank materials made necessary by changing environmental requirements. Chemical fingerprinting can detect and diagnose variations in material composition. To chemically characterize each material, fingerprint methods are selected from an extensive toolbox based on the material's chemistry and the ability of the specific methods to detect the material's critical ingredients. Fingerprint methods have been developed for a variety of materials including Thermal Protection System foams, adhesives, primers, and composites.

Combining in situ characterization methods in one set-up: looking with more eyes into the intricate chemistry of the synthesis and working of heterogeneous catalysts.

PubMed

Bentrup, Ursula

2010-12-01

Several in situ techniques are known which allow investigations of catalysts and catalytic reactions under real reaction conditions using different spectroscopic and X-ray methods. In recent years, specific set-ups have been established which combine two or more in situ methods in order to get a more detailed understanding of catalytic systems. This tutorial review will give a summary of currently available set-ups equipped with multiple techniques for in situ catalyst characterization, catalyst preparation, and reaction monitoring. Besides experimental and technical aspects of method coupling including X-ray techniques, spectroscopic methods (Raman, UV-vis, FTIR), and magnetic resonance spectroscopies (NMR, EPR), essential results will be presented to demonstrate the added value of multitechnique in situ approaches. A special section is focussed on selected examples of use which show new developments and application fields.

Isolation and characterization of anti-SEB peptides using magnetic sorting and bacterial peptide display library technology

NASA Astrophysics Data System (ADS)

Pennington, Joseph M.; Kogot, Joshua M.; Sarkes, Deborah A.; Pellegrino, Paul M.; Stratis-Cullum, Dimitra N.

2012-06-01

Peptide display libraries offer an alternative method to existing antibody development methods enabling rapid isolation of highly stable reagents for detection of new and emerging biological threats. Bacterial display libraries are used to isolate new peptide reagents within 1 week, which is simpler and timelier than using competing display library technology based on phage or yeast. Using magnetic sorting methods, we have isolated peptide reagents with high affinity and specificity to staphylococcal enterotoxin B (SEB), a suspected food pathogen. Flow cytometry methods were used for on-cell characterization and the binding affinity (Kd) of this new peptide reagent was determined to be 56 nm with minimal cross-reactivity to other proteins. These results demonstrated that magnetic sorting for new reagents using bacterial display libraries is a rapid and effective method and has the potential for current and new and emerging food pathogen targets.

Selenium-regulated hierarchy of human selenoproteome in cancerous and immortalized cells lines.

PubMed

Touat-Hamici, Zahia; Bulteau, Anne-Laure; Bianga, Juliusz; Jean-Jacques, Hélène; Szpunar, Joanna; Lobinski, Ryszard; Chavatte, Laurent

2018-04-13

Selenoproteins (25 genes in human) co-translationally incorporate selenocysteine using a UGA codon, normally used as a stop signal. The human selenoproteome is primarily regulated by selenium bioavailability with a tissue-specific hierarchy. We investigated the hierarchy of selenoprotein expression in response to selenium concentration variation in four cell lines originating from kidney (HEK293, immortalized), prostate (LNCaP, cancer), skin (HaCaT, immortalized) and liver (HepG2, cancer), using complementary analytical methods. We performed (i) enzymatic activity, (ii) RT-qPCR, (iii) immuno-detection, (iv) selenium-specific mass spectrometric detection after non-radioactive 76 Se labeling of selenoproteins, and (v) luciferase-based reporter constructs in various cell extracts. We characterized cell-line specific alterations of the selenoproteome in response to selenium variation that, in most of the cases, resulted from a translational control of gene expression. We established that UGA-selenocysteine recoding efficiency, which depends on the nature of the SECIS element, dictates the response to selenium variation. We characterized that selenoprotein hierarchy is cell-line specific with conserved features. This analysis should be done prior to any experiments in a novel cell line. We reported a strategy based on complementary methods to evaluate selenoproteome regulation in human cells in culture. Copyright © 2018 Elsevier B.V. All rights reserved.

A Robust Static Headspace GC-FID Method to Detect and Quantify Formaldehyde Impurity in Pharmaceutical Excipients

PubMed Central

Al-Khayat, Mohammad Ammar; Karabet, Francois; Al-Mardini, Mohammad Amer

2018-01-01

Formaldehyde is a highly reactive impurity that can be found in many pharmaceutical excipients. Trace levels of this impurity may affect drug product stability, safety, efficacy, and performance. A static headspace gas chromatographic method was developed and validated to determine formaldehyde in pharmaceutical excipients after an effective derivatization procedure using acidified ethanol. Diethoxymethane, the derivative of formaldehyde, was then directly analyzed by GC-FID. Despite the simplicity of the developed method, however, it is characterized by its specificity, accuracy, and precision. The limits of detection and quantification of formaldehyde in the samples were of 2.44 and 8.12 µg/g, respectively. This method is characterized by using simple and economic GC-FID technique instead of MS detection, and it is successfully used to analyze formaldehyde in commonly used pharmaceutical excipients. PMID:29686930

Investigating the Functional Role of Prostate-Specific Membrane Antigen and its Enzymatic Activity in Prostate Cancer Metastasis

DTIC Science & Technology

2008-02-01

fluorescent probes for live cell imaging . PSMA distribution of cells grown on different extracellular matrices will be characterized to provide guidance...PCa migration, using in vitro cell model systems and live - cell imaging methods, we characterized the role of PSMA in cell motility and adhesion. Using...Generated fluorescently conjugated anti-PSMA antibodies for live cell imaging . 2. Optimized the siRNA-PSMA transfection and achieved an approximately

Photometric Application of the Gram Stain Method To Characterize Natural Bacterial Populations in Aquatic Environments

PubMed Central

Saida, H.; Ytow, N.; Seki, H.

1998-01-01

The Gram stain method was applied to the photometric characterization of aquatic bacterial populations with a charge-coupled device camera and an image analyzer. Escherichia coli and Bacillus subtilis were used as standards of typical gram-negative and gram-positive bacteria, respectively. A mounting agent to obtain clear images of Gram-stained bacteria on Nuclepore membrane filters was developed. The bacterial stainability by the Gram stain was indicated by the Gram stain index (GSI), which was applicable not only to the dichotomous classification of bacteria but also to the characterization of cell wall structure. The GSI spectra of natural bacterial populations in water with various levels of eutrophication showed a distinct profile, suggesting possible staining specificity that indicates the presence of a particular bacterial population in the aquatic environment. PMID:9464416

Study on the evaluation method for fault displacement based on characterized source model

NASA Astrophysics Data System (ADS)

Tonagi, M.; Takahama, T.; Matsumoto, Y.; Inoue, N.; Irikura, K.; Dalguer, L. A.

2016-12-01

In IAEA Specific Safety Guide (SSG) 9 describes that probabilistic methods for evaluating fault displacement should be used if no sufficient basis is provided to decide conclusively that the fault is not capable by using the deterministic methodology. In addition, International Seismic Safety Centre compiles as ANNEX to realize seismic hazard for nuclear facilities described in SSG-9 and shows the utility of the deterministic and probabilistic evaluation methods for fault displacement. In Japan, it is required that important nuclear facilities should be established on ground where fault displacement will not arise when earthquakes occur in the future. Under these situations, based on requirements, we need develop evaluation methods for fault displacement to enhance safety in nuclear facilities. We are studying deterministic and probabilistic methods with tentative analyses using observed records such as surface fault displacement and near-fault strong ground motions of inland crustal earthquake which fault displacements arose. In this study, we introduce the concept of evaluation methods for fault displacement. After that, we show parts of tentative analysis results for deterministic method as follows: (1) For the 1999 Chi-Chi earthquake, referring slip distribution estimated by waveform inversion, we construct a characterized source model (Miyake et al., 2003, BSSA) which can explain observed near-fault broad band strong ground motions. (2) Referring a characterized source model constructed in (1), we study an evaluation method for surface fault displacement using hybrid method, which combines particle method and distinct element method. At last, we suggest one of the deterministic method to evaluate fault displacement based on characterized source model. This research was part of the 2015 research project `Development of evaluating method for fault displacement` by the Secretariat of Nuclear Regulation Authority (S/NRA), Japan.

Reconstructing the Genomic Content of Microbiome Taxa through Shotgun Metagenomic Deconvolution

PubMed Central

Carr, Rogan; Shen-Orr, Shai S.; Borenstein, Elhanan

2013-01-01

Metagenomics has transformed our understanding of the microbial world, allowing researchers to bypass the need to isolate and culture individual taxa and to directly characterize both the taxonomic and gene compositions of environmental samples. However, associating the genes found in a metagenomic sample with the specific taxa of origin remains a critical challenge. Existing binning methods, based on nucleotide composition or alignment to reference genomes allow only a coarse-grained classification and rely heavily on the availability of sequenced genomes from closely related taxa. Here, we introduce a novel computational framework, integrating variation in gene abundances across multiple samples with taxonomic abundance data to deconvolve metagenomic samples into taxa-specific gene profiles and to reconstruct the genomic content of community members. This assembly-free method is not bounded by various factors limiting previously described methods of metagenomic binning or metagenomic assembly and represents a fundamentally different approach to metagenomic-based genome reconstruction. An implementation of this framework is available at http://elbo.gs.washington.edu/software.html. We first describe the mathematical foundations of our framework and discuss considerations for implementing its various components. We demonstrate the ability of this framework to accurately deconvolve a set of metagenomic samples and to recover the gene content of individual taxa using synthetic metagenomic samples. We specifically characterize determinants of prediction accuracy and examine the impact of annotation errors on the reconstructed genomes. We finally apply metagenomic deconvolution to samples from the Human Microbiome Project, successfully reconstructing genus-level genomic content of various microbial genera, based solely on variation in gene count. These reconstructed genera are shown to correctly capture genus-specific properties. With the accumulation of metagenomic data, this deconvolution framework provides an essential tool for characterizing microbial taxa never before seen, laying the foundation for addressing fundamental questions concerning the taxa comprising diverse microbial communities. PMID:24146609

Etching-free patterning method for electrical characterization of atomically thin MoSe2 films grown by chemical vapor deposition

NASA Astrophysics Data System (ADS)

Utama, M. Iqbal Bakti; Lu, Xin; Zhan, Da; Ha, Son Tung; Yuan, Yanwen; Shen, Zexiang; Xiong, Qihua

2014-10-01

Patterning two-dimensional materials into specific spatial arrangements and geometries is essential for both fundamental studies of materials and practical applications in electronics. However, the currently available patterning methods generally require etching steps that rely on complicated and expensive procedures. We report here a facile patterning method for atomically thin MoSe2 films using stripping with an SU-8 negative resist layer exposed to electron beam lithography. Additional steps of chemical and physical etching were not necessary in this SU-8 patterning method. The SU-8 patterning was used to define a ribbon channel from a field effect transistor of MoSe2 film, which was grown by chemical vapor deposition. The narrowing of the conduction channel area with SU-8 patterning was crucial in suppressing the leakage current within the device, thereby allowing a more accurate interpretation of the electrical characterization results from the sample. An electrical transport study, enabled by the SU-8 patterning, showed a variable range hopping behavior at high temperatures.Patterning two-dimensional materials into specific spatial arrangements and geometries is essential for both fundamental studies of materials and practical applications in electronics. However, the currently available patterning methods generally require etching steps that rely on complicated and expensive procedures. We report here a facile patterning method for atomically thin MoSe2 films using stripping with an SU-8 negative resist layer exposed to electron beam lithography. Additional steps of chemical and physical etching were not necessary in this SU-8 patterning method. The SU-8 patterning was used to define a ribbon channel from a field effect transistor of MoSe2 film, which was grown by chemical vapor deposition. The narrowing of the conduction channel area with SU-8 patterning was crucial in suppressing the leakage current within the device, thereby allowing a more accurate interpretation of the electrical characterization results from the sample. An electrical transport study, enabled by the SU-8 patterning, showed a variable range hopping behavior at high temperatures. Electronic supplementary information (ESI) available: Further experiments on patterning and additional electrical characterizations data. See DOI: 10.1039/c4nr03817g

Method development and validation for measuring the particle size distribution of pentaerythritol tetranitrate (PETN) powders.

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

Young, Sharissa Gay

2005-09-01

Currently, the critical particle properties of pentaerythritol tetranitrate (PETN) that influence deflagration-to-detonation time in exploding bridge wire detonators (EBW) are not known in sufficient detail to allow development of a predictive failure model. The specific surface area (SSA) of many PETN powders has been measured using both permeametry and gas absorption methods and has been found to have a critical effect on EBW detonator performance. The permeametry measure of SSA is a function of particle shape, packed bed pore geometry, and particle size distribution (PSD). Yet there is a general lack of agreement in PSD measurements between laboratories, raising concernsmore » regarding collaboration and complicating efforts to understand changes in EBW performance related to powder properties. Benchmarking of data between laboratories that routinely perform detailed PSD characterization of powder samples and the determination of the most appropriate method to measure each PETN powder are necessary to discern correlations between performance and powder properties and to collaborate with partnering laboratories. To this end, a comparison was made of the PSD measured by three laboratories using their own standard procedures for light scattering instruments. Three PETN powder samples with different surface areas and particle morphologies were characterized. Differences in bulk PSD data generated by each laboratory were found to result from variations in sonication of the samples during preparation. The effect of this sonication was found to depend on particle morphology of the PETN samples, being deleterious to some PETN samples and advantageous for others in moderation. Discrepancies in the submicron-sized particle characterization data were related to an instrument-specific artifact particular to one laboratory. The type of carrier fluid used by each laboratory to suspend the PETN particles for the light scattering measurement had no consistent effect on the resulting PSD data. Finally, the SSA of the three powders was measured using both permeametry and gas absorption methods, enabling the PSD to be linked to the SSA for these PETN powders. Consistent characterization of other PETN powders can be performed using the appropriate sample-specific preparation method, so that future studies can accurately identify the effect of changes in the PSD on the SSA and ultimately model EBW performance.« less

A Review of Interface Electronic Systems for AT-cut Quartz Crystal Microbalance Applications in Liquids

PubMed Central

Arnau, Antonio

2008-01-01

From the first applications of AT-cut quartz crystals as sensors in solutions more than 20 years ago, the so-called quartz crystal microbalance (QCM) sensor is becoming into a good alternative analytical method in a great deal of applications such as biosensors, analysis of biomolecular interactions, study of bacterial adhesion at specific interfaces, pathogen and microorganism detection, study of polymer film-biomolecule or cell-substrate interactions, immunosensors and an extensive use in fluids and polymer characterization and electrochemical applications among others. The appropriate evaluation of this analytical method requires recognizing the different steps involved and to be conscious of their importance and limitations. The first step involved in a QCM system is the accurate and appropriate characterization of the sensor in relation to the specific application. The use of the piezoelectric sensor in contact with solutions strongly affects its behavior and appropriate electronic interfaces must be used for an adequate sensor characterization. Systems based on different principles and techniques have been implemented during the last 25 years. The interface selection for the specific application is important and its limitations must be known to be conscious of its suitability, and for avoiding the possible error propagation in the interpretation of results. This article presents a comprehensive overview of the different techniques used for AT-cut quartz crystal microbalance in in-solution applications, which are based on the following principles: network or impedance analyzers, decay methods, oscillators and lock-in techniques. The electronic interfaces based on oscillators and phase-locked techniques are treated in detail, with the description of different configurations, since these techniques are the most used in applications for detection of analytes in solutions, and in those where a fast sensor response is necessary. PMID:27879713

Validation of a thin-layer chromatography/densitometry method for the characterization of invertase activity.

PubMed

Ferey, Justine; Da Silva, David; Bravo-Veyrat, Sophie; Lafite, Pierre; Daniellou, Richard; Maunit, Benoît

2016-12-16

This paper presents a kinetic study of invertase, a specific fructofuranosidase cloned from the Leishmania major genome. The kinetic parameters of the β-d-fructofuranosidase from Leishmania major (BfrA) were determined using Thin-Layer Chromatography (TLC) and UV-densitometry (TLC@UV) specifically developed for the separation and detection of three carbohydrates namely sucrose, glucose and fructose. Separation was performed on TLC silica gel 60 F254 plates impregnated with sodium bisulphate and citrate and heated prior to development. This fast and easy separation was performed with two successive developments using ACN/H 2 O 80/20 (v/v) as mobile phase. Sensitive and repeatable derivatization of sugars was achieved by dipping the plates in a solution of 4-aminobenzoic acid. Quantification was performed by UV-detection. The method was validated according to ICH guidelines Q2(R1) in terms of specificity, limits of detection and quantification, precision and robustness (with n=3 replicates and CV ≤10%). The characterization of BfrA reaction kinetic was performed by monitoring the accumulation of either glucose or fructose detected by TLC@UV. Hydrolysis of sucrose was described by the Michaelis-Menten kinetic parameters (K M ; V max ) respectively equal to 63.09±7.590mM; 0.037±0.00094mM/min using glucose production and 83.01±14.39mM; 0.031±0.0021mM/min monitoring fructose. Hydrolyses of three alternative substrates, raffinose, stachyose and inulin, were also compared and the regiospecificity of the reaction was characterized. This TLC@UV method is shown to be suitable for the refined kinetic analysis of different reactions related to the hydrolysis of sugars. Copyright © 2016. Published by Elsevier B.V.

Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE

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

Pidatala, Venkataramana R.; Mahboubi, Amir; Mortimer, Jenny C.

Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. Gels can be visualized with a modified gel imaging system (see Table of Materials). The resulting oligosaccharidemore » fingerprint can either be compared qualitatively or, with replication, quantitatively. Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates.« less

Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE.

PubMed

Pidatala, Venkataramana R; Mahboubi, Amir; Mortimer, Jenny C

2017-10-16

Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. Gels can be visualized with a modified gel imaging system (see Table of Materials). The resulting oligosaccharide fingerprint can either be compared qualitatively or, with replication, quantitatively. Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates.

Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE

DOE PAGES

Pidatala, Venkataramana R.; Mahboubi, Amir; Mortimer, Jenny C.

2017-10-16

Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. Gels can be visualized with a modified gel imaging system (see Table of Materials). The resulting oligosaccharidemore » fingerprint can either be compared qualitatively or, with replication, quantitatively. Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates.« less

Probabilistic Exposure Analysis for Chemical Risk Characterization

PubMed Central

Bogen, Kenneth T.; Cullen, Alison C.; Frey, H. Christopher; Price, Paul S.

2009-01-01

This paper summarizes the state of the science of probabilistic exposure assessment (PEA) as applied to chemical risk characterization. Current probabilistic risk analysis methods applied to PEA are reviewed. PEA within the context of risk-based decision making is discussed, including probabilistic treatment of related uncertainty, interindividual heterogeneity, and other sources of variability. Key examples of recent experience gained in assessing human exposures to chemicals in the environment, and other applications to chemical risk characterization and assessment, are presented. It is concluded that, although improvements continue to be made, existing methods suffice for effective application of PEA to support quantitative analyses of the risk of chemically induced toxicity that play an increasing role in key decision-making objectives involving health protection, triage, civil justice, and criminal justice. Different types of information required to apply PEA to these different decision contexts are identified, and specific PEA methods are highlighted that are best suited to exposure assessment in these separate contexts. PMID:19223660

Label-free resistive-pulse cytometry.

PubMed

Chapman, M R; Sohn, L L

2011-01-01

Numerous methods have recently been developed to characterize cells for size, shape, and specific cell-surface markers. Most of these methods rely upon exogenous labeling of the cells and are better suited for large cell populations (>10,000). Here, we review a label-free method of characterizing and screening cells based on the Coulter-counter technique of particle sizing: an individual cell transiting a microchannel (or "pore") causes a downward pulse in the measured DC current across that "pore". Pulse magnitude corresponds to the cell size, pulse width to the transit time needed for the cell to pass through the pore, and pulse shape to how the cell traverses across the pore (i.e., rolling or tumbling). When the pore is functionalized with an antibody that is specific to a surface-epitope of interest, label-free screening of a specific marker is possible, as transient binding between the two results in longer time duration than when the pore is unfunctionalized or functionalized with a nonspecific antibody. While this method cannot currently compete with traditional technology in terms of throughput, there are a number of applications for which this technology is better suited than current commercial cytometry systems. Applications include the rapid and nondestructive analysis of small cell populations (<100), which is not possible with current technology, and a platform for providing true point-of-care clinical diagnostics, due to the simplicity of the device, low manufacturing costs, and ease of use. Copyright © 2011 Elsevier Inc. All rights reserved.

Methods for Characterization of Alternative RNA Splicing.

PubMed

Harvey, Samuel E; Cheng, Chonghui

2016-01-01

Quantification of alternative splicing to detect the abundance of differentially spliced isoforms of a gene in total RNA can be accomplished via RT-PCR using both quantitative real-time and semi-quantitative PCR methods. These methods require careful PCR primer design to ensure specific detection of particular splice isoforms. We also describe analysis of alternative splicing using a splicing "minigene" in mammalian cell tissue culture to facilitate investigation of the regulation of alternative splicing of a particular exon of interest.

Detection and Characterization of Reactive Oxygen and Nitrogen Species in Biological Systems by Monitoring Species-Specific Products.

PubMed

Hardy, Micael; Zielonka, Jacek; Karoui, Hakim; Sikora, Adam; Michalski, Radosław; Podsiadły, Radosław; Lopez, Marcos; Vasquez-Vivar, Jeannette; Kalyanaraman, Balaraman; Ouari, Olivier

2018-05-20

Since the discovery of the superoxide dismutase enzyme, the generation and fate of short-lived oxidizing, nitrosating, nitrating, and halogenating species in biological systems has been of great interest. Despite the significance of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in numerous diseases and intracellular signaling, the rigorous detection of ROS and RNS has remained a challenge. Recent Advances: Chemical characterization of the reactions of selected ROS and RNS with electron paramagnetic resonance (EPR) spin traps and fluorescent probes led to the establishment of species-specific products, which can be used for specific detection of several forms of ROS and RNS in cell-free systems and in cultured cells in vitro and in animals in vivo. Profiling oxidation products from the ROS and RNS probes provides a rigorous method for detection of those species in biological systems. Formation and detection of species-specific products from the probes enables accurate characterization of the oxidative environment in cells. Measurement of the total signal (fluorescence, chemiluminescence, etc.) intensity does not allow for identification of the ROS/RNS formed. It is critical to identify the products formed by using chromatographic or other rigorous techniques. Product analyses should be accompanied by monitoring of the intracellular probe level, another factor controlling the yield of the product(s) formed. More work is required to characterize the chemical reactivity of the ROS/RNS probes, and to develop new probes/detection approaches enabling real-time, selective monitoring of the specific products formed from the probes. Antioxid. Redox Signal. 28, 1416-1432.

Characterization of Protein-Excipient Microheterogeneity in Biopharmaceutical Solid-State Formulations by Confocal Fluorescence Microscopy.

PubMed

Koshari, Stijn H S; Ross, Jean L; Nayak, Purnendu K; Zarraga, Isidro E; Rajagopal, Karthikan; Wagner, Norman J; Lenhoff, Abraham M

2017-02-06

Protein-stabilizer microheterogeneity is believed to influence long-term protein stability in solid-state biopharmaceutical formulations and its characterization is therefore essential for the rational design of stable formulations. However, the spatial distribution of the protein and the stabilizer in a solid-state formulation is, in general, difficult to characterize because of the lack of a functional, simple, and reliable characterization technique. We demonstrate the use of confocal fluorescence microscopy with fluorescently labeled monoclonal antibodies (mAbs) and antibody fragments (Fabs) to directly visualize three-dimensional particle morphologies and protein distributions in dried biopharmaceutical formulations, without restrictions on processing conditions or the need for extensive data analysis. While industrially relevant lyophilization procedures of a model IgG1 mAb generally lead to uniform protein-excipient distribution, the method shows that specific spray-drying conditions lead to distinct protein-excipient segregation. Therefore, this method can enable more definitive optimization of formulation conditions than has previously been possible.

Advanced eddy current test signal analysis for steam generator tube defect classification and characterization

NASA Astrophysics Data System (ADS)

McClanahan, James Patrick

Eddy Current Testing (ECT) is a Non-Destructive Examination (NDE) technique that is widely used in power generating plants (both nuclear and fossil) to test the integrity of heat exchanger (HX) and steam generator (SG) tubing. Specifically for this research, laboratory-generated, flawed tubing data were examined. The purpose of this dissertation is to develop and implement an automated method for the classification and an advanced characterization of defects in HX and SG tubing. These two improvements enhanced the robustness of characterization as compared to traditional bobbin-coil ECT data analysis methods. A more robust classification and characterization of the tube flaw in-situ (while the SG is on-line but not when the plant is operating), should provide valuable information to the power industry. The following are the conclusions reached from this research. A feature extraction program acquiring relevant information from both the mixed, absolute and differential data was successfully implemented. The CWT was utilized to extract more information from the mixed, complex differential data. Image Processing techniques used to extract the information contained in the generated CWT, classified the data with a high success rate. The data were accurately classified, utilizing the compressed feature vector and using a Bayes classification system. An estimation of the upper bound for the probability of error, using the Bhattacharyya distance, was successfully applied to the Bayesian classification. The classified data were separated according to flaw-type (classification) to enhance characterization. The characterization routine used dedicated, flaw-type specific ANNs that made the characterization of the tube flaw more robust. The inclusion of outliers may help complete the feature space so that classification accuracy is increased. Given that the eddy current test signals appear very similar, there may not be sufficient information to make an extremely accurate (>95%) classification or an advanced characterization using this system. It is necessary to have a larger database fore more accurate system learning.

Analytical methods for characterization of explosives-contaminated sites on U.S. Army installations

NASA Astrophysics Data System (ADS)

Jenkins, Thomas F.; Walsh, Marianne E.; Thorne, Philip G.

1995-10-01

The U.S. Army manufactures munitions at facilities throughout the United States. Many of these facilities are contaminated with residues of explosives from production, disposal of off- specification, and out-of-data munitions. The first step in remediating these sites is careful characterization. Currently sites are being characterized using a combination of on-site field screening and off-site laboratory analysis. Most of the contamination is associated with TNT (2,4,6-trinitrotoluene) and RDX (hexahydro-1,3,5-tri-nitro-1,3,5-triazine) and their manufacturing impurities and environmental transformation products. Both colorimetric and enzyme immunoassay-based field screening methods have been used successfully for on-site characterization. These methods have similar detection capabilities but differ in their selectivity. Although field screening is very cost-effective, laboratory analysis is still required to fully characterize a site. Laboratory analysis for explosives residues in the United States is generally conducted using high-performance liquid chromatography equipped with a UV detector. Air-dried soils are extracted with acetonitrile in an ultrasonic bath. Water is analyzed directly if detection limits in the range of 10 - 20 (mu) g/L are acceptable, or preconcentrated using either salting-out solvent extraction with acetonitrile or solid phase extraction.

Modified Standard Penetration Test–based Drilled Shaft Design Method for Weak Rocks (Phase 2 Study)

DOT National Transportation Integrated Search

2017-12-15

In this project, Illinois-specific design procedures were developed for drilled shafts founded in weak shale or rock. In particular, a modified standard penetration test was developed and verified to characterize the in situ condition of weak shales ...

ISO 15859 Propellant and Fluid Specifications: A Review and Comparison with Military and NASA Specifications

NASA Technical Reports Server (NTRS)

Greene, Ben; McClure, Mark B.; Baker, David L.

2006-01-01

This work presents an overview of the International Organization for Standardization (ISO) 15859 International Standard for Space Systems Fluid Characteristics, Sampling and Test Methods Parts 1 through 13 issued in June 2004. These standards establish requirements for fluid characteristics, sampling, and test methods for 13 fluids of concern to the propellant community and propellant characterization laboratories: oxygen, hydrogen, nitrogen, helium, nitrogen tetroxide, monomethylhydrazine, hydrazine, kerosene, argon, water, ammonia, carbon dioxide, and breathing air. A comparison of the fluid characteristics, sampling, and test methods required by the ISO standards to the current military and NASA specifications, which are in use at NASA facilities and elsewhere, is presented. Many ISO standards composition limits and other content agree with those found in the applicable parts of NASA SE-S-0073, NASA SSP 30573, military performance standards and details, and Compressed Gas Association (CGA) commodity specifications. The status of a current project managed at NASA Johnson Space Center White Sands Test Facility (WSTF) to rewrite these documents is discussed.

Comparing biomarker measurements to a normal range: when ...

EPA Pesticide Factsheets

This commentary is the second of a series outlining one specific concept in interpreting biomarkers data. In the first, an observational method was presented for assessing the distribution of measurements before making parametric calculations. Here, the discussion revolves around the next step, the choice of using standard error of the mean or the calculated standard deviation to compare or predict measurement results. The National Exposure Research Laboratory’s (NERL’s) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA’s mission to protect human health and the environment. HEASD’s research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA’s strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.

A bioinformatics approach for identifying transgene insertion sites using whole genome sequencing data.

PubMed

Park, Doori; Park, Su-Hyun; Ban, Yong Wook; Kim, Youn Shic; Park, Kyoung-Cheul; Kim, Nam-Soo; Kim, Ju-Kon; Choi, Ik-Young

2017-08-15

Genetically modified crops (GM crops) have been developed to improve the agricultural traits of modern crop cultivars. Safety assessments of GM crops are of paramount importance in research at developmental stages and before releasing transgenic plants into the marketplace. Sequencing technology is developing rapidly, with higher output and labor efficiencies, and will eventually replace existing methods for the molecular characterization of genetically modified organisms. To detect the transgenic insertion locations in the three GM rice gnomes, Illumina sequencing reads are mapped and classified to the rice genome and plasmid sequence. The both mapped reads are classified to characterize the junction site between plant and transgene sequence by sequence alignment. Herein, we present a next generation sequencing (NGS)-based molecular characterization method, using transgenic rice plants SNU-Bt9-5, SNU-Bt9-30, and SNU-Bt9-109. Specifically, using bioinformatics tools, we detected the precise insertion locations and copy numbers of transfer DNA, genetic rearrangements, and the absence of backbone sequences, which were equivalent to results obtained from Southern blot analyses. NGS methods have been suggested as an effective means of characterizing and detecting transgenic insertion locations in genomes. Our results demonstrate the use of a combination of NGS technology and bioinformatics approaches that offers cost- and time-effective methods for assessing the safety of transgenic plants.

Has the use of molecular methods for the characterization of the human oral microbiome changed our understanding of the role of bacteria in the pathogenesis of periodontal disease?

PubMed

Wade, William Geoffrey

2011-03-01

Only around half of oral bacteria can be grown in the laboratory using conventional culture methods. Molecular methods based on 16S rRNA gene sequence are now available and are being used to characterize the periodontal microbiota in its entirety. This review describes the cultural characterization of the oral and periodontal microbiotas and explores the influence of the additional data now available from culture-independent molecular analyses on current thinking on the role of bacteria in periodontitis. Culture-independent molecular analysis of the periodontal microbiota has shown it to be far more diverse than previously thought. A number of species including some that have yet to be cultured are as strongly associated with disease as those organisms traditionally regarded as periodontal pathogens. Sequencing of bacterial genomes has revealed a high degree of intra-specific genetic diversity. The use of molecular methods for the characterization of the periodontal microbiome has greatly expanded the range of bacterial species known to colonize this habitat. Understanding the interactions between the human host and its commensal bacterial community at the functional level is a priority. © 2011 John Wiley & Sons A/S.

Exploitation of the Ornithine Effect Enhances Characterization of Stapled and Cyclic Peptides

NASA Astrophysics Data System (ADS)

Crittenden, Christopher M.; Parker, W. Ryan; Jenner, Zachary B.; Bruns, Kerry A.; Akin, Lucas D.; McGee, William M.; Ciccimaro, Eugene; Brodbelt, Jennifer S.

2016-05-01

A method to facilitate the characterization of stapled or cyclic peptides is reported via an arginine-selective derivatization strategy coupled with MS/MS analysis. Arginine residues are converted to ornithine residues through a deguanidination reaction that installs a highly selectively cleavable site in peptides. Upon activation by CID or UVPD, the ornithine residue cyclizes to promote cleavage of the adjacent amide bond. This Arg-specific process offers a unique strategy for site-selective ring opening of stapled and cyclic peptides. Upon activation of each derivatized peptide, site-specific backbone cleavage at the ornithine residue results in two complementary products: the lactam ring-containing portion of the peptide and the amine-containing portion. The deguanidination process not only provides a specific marker site that initiates fragmentation of the peptide but also offers a means to unlock the staple and differentiate isobaric stapled peptides.

PM2.5 Technology Assessment and Characterization Study in New York - PMTACS-NY: The 2001 Summer Field Intensive in Queens, NY

NASA Astrophysics Data System (ADS)

Demerjian, K. L.

2002-12-01

In the summer of 2001, an intensive field measurement campaign was carried out in Queens, NY as part of the PM2.5 Technology Assessment and Characterization Study in New York (PMTACS-NY) to characterize the physical and chemical composition of particulate matter and related precursors utilizing conventional and advanced instrumentation technologies. The measurement program, involving a team of scientists from federal, state, university and private sector organizations, was designed to provide detailed time resolved chemical and physical characterization of the urban PM2.5/co-pollutant complex in relation to the regional environment. A summary of the chemical and meteorological data defining specific events during the field intensive is presented as are results addressing specific hypothesis designed around PMTACS-NY program objectives. These include initial findings and conclusions related to 1) performance testing and evaluation of emerging measurement technologies and comparison with EPA mandated PM federal reference methods currently operational as part of the New York State and national PM2.5 monitoring network; 2) emissions characterization of CNG, standard diesel and CRT (Continuously Regenerating Technology) diesel retrofit powered vehicles; and 3) compositional comparisons of urban and regional PM2.5.

A hybrid framework for reservoir characterization using fuzzy ranking and an artificial neural network

NASA Astrophysics Data System (ADS)

Wang, Baijie; Wang, Xin; Chen, Zhangxin

2013-08-01

Reservoir characterization refers to the process of quantitatively assigning reservoir properties using all available field data. Artificial neural networks (ANN) have recently been introduced to solve reservoir characterization problems dealing with the complex underlying relationships inherent in well log data. Despite the utility of ANNs, the current limitation is that most existing applications simply focus on directly implementing existing ANN models instead of improving/customizing them to fit the specific reservoir characterization tasks at hand. In this paper, we propose a novel intelligent framework that integrates fuzzy ranking (FR) and multilayer perceptron (MLP) neural networks for reservoir characterization. FR can automatically identify a minimum subset of well log data as neural inputs, and the MLP is trained to learn the complex correlations from the selected well log data to a target reservoir property. FR guarantees the selection of the optimal subset of representative data from the overall well log data set for the characterization of a specific reservoir property; and, this implicitly improves the modeling and predication accuracy of the MLP. In addition, a growing number of industrial agencies are implementing geographic information systems (GIS) in field data management; and, we have designed the GFAR solution (GIS-based FR ANN Reservoir characterization solution) system, which integrates the proposed framework into a GIS system that provides an efficient characterization solution. Three separate petroleum wells from southwestern Alberta, Canada, were used in the presented case study of reservoir porosity characterization. Our experiments demonstrate that our method can generate reliable results.

Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy.

PubMed

Friedrichs, Stephanie; Malan, Daniela; Voss, Yvonne; Sasse, Philipp

2015-01-08

Disease-specific induced pluripotent stem (iPS) cells can be generated from patients and differentiated into functional cardiomyocytes for characterization of the disease and for drug screening. In order to obtain pure cardiomyocytes for automated electrophysiological investigation, we here report a novel non-clonal purification strategy by using lentiviral gene transfer of a puromycin resistance gene under the control of a cardiac-specific promoter. We have applied this method to our previous reported wild-type and long QT syndrome 3 (LQTS 3)-specific mouse iPS cells and obtained a pure cardiomyocyte population. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. Action potentials and field potentials showed the characteristic prolongation at low heart rates in LQTS 3-specific, but not in wild-type iPS cell-derived cardiomyocytes. Hence, LQTS 3-specific cardiomyocytes can be purified from iPS cells with a lentiviral strategy, maintain the hallmarks of the LQTS 3 disease and can be used for automated electrophysiological characterization and drug screening.

Surface characterization of nanomaterials and nanoparticles: Important needs and challenging opportunities

PubMed Central

Baer, Donald R.; Engelhard, Mark H.; Johnson, Grant E.; Laskin, Julia; Lai, Jinfeng; Mueller, Karl; Munusamy, Prabhakaran; Thevuthasan, Suntharampillai; Wang, Hongfei; Washton, Nancy; Elder, Alison; Baisch, Brittany L.; Karakoti, Ajay; Kuchibhatla, Satyanarayana V. N. T.; Moon, DaeWon

2013-01-01

This review examines characterization challenges inherently associated with understanding nanomaterials and the roles surface and interface characterization methods can play in meeting some of the challenges. In parts of the research community, there is growing recognition that studies and published reports on the properties and behaviors of nanomaterials often have reported inadequate or incomplete characterization. As a consequence, the true value of the data in these reports is, at best, uncertain. With the increasing importance of nanomaterials in fundamental research and technological applications, it is desirable that researchers from the wide variety of disciplines involved recognize the nature of these often unexpected challenges associated with reproducible synthesis and characterization of nanomaterials, including the difficulties of maintaining desired materials properties during handling and processing due to their dynamic nature. It is equally valuable for researchers to understand how characterization approaches (surface and otherwise) can help to minimize synthesis surprises and to determine how (and how quickly) materials and properties change in different environments. Appropriate application of traditional surface sensitive analysis methods (including x-ray photoelectron and Auger electron spectroscopies, scanning probe microscopy, and secondary ion mass spectroscopy) can provide information that helps address several of the analysis needs. In many circumstances, extensions of traditional data analysis can provide considerably more information than normally obtained from the data collected. Less common or evolving methods with surface selectivity (e.g., some variations of nuclear magnetic resonance, sum frequency generation, and low and medium energy ion scattering) can provide information about surfaces or interfaces in working environments (operando or in situ) or information not provided by more traditional methods. Although these methods may require instrumentation or expertise not generally available, they can be particularly useful in addressing specific questions, and examples of their use in nanomaterial research are presented. PMID:24482557

Surface characterization of nanomaterials and nanoparticles: Important needs and challenging opportunities

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

Baer, Donald R.; Engelhard, Mark H.; Johnson, Grant E.

2013-09-15

This review examines characterization challenges inherently associated with understanding nanomaterials and the roles surface and interface characterization methods can play in meeting some of the challenges. In parts of the research community, there is growing recognition that studies and published reports on the properties and behaviors of nanomaterials often have reported inadequate or incomplete characterization. As a consequence, the true value of the data in these reports is, at best, uncertain. With the increasing importance of nanomaterials in fundamental research and technological applications, it is desirable that researchers from the wide variety of disciplines involved recognize the nature of thesemore » often unexpected challenges associated with reproducible synthesis and characterization of nanomaterials, including the difficulties of maintaining desired materials properties during handling and processing due to their dynamic nature. It is equally valuable for researchers to understand how characterization approaches (surface and otherwise) can help to minimize synthesis surprises and to determine how (and how quickly) materials and properties change in different environments. Appropriate application of traditional surface sensitive analysis methods (including x-ray photoelectron and Auger electron spectroscopies, scanning probe microscopy, and secondary ion mass spectroscopy) can provide information that helps address several of the analysis needs. In many circumstances, extensions of traditional data analysis can provide considerably more information than normally obtained from the data collected. Less common or evolving methods with surface selectivity (e.g., some variations of nuclear magnetic resonance, sum frequency generation, and low and medium energy ion scattering) can provide information about surfaces or interfaces in working environments (operando or in situ) or information not provided by more traditional methods. Although these methods may require instrumentation or expertise not generally available, they can be particularly useful in addressing specific questions, and examples of their use in nanomaterial research are presented.« less

Solar Technology Validation Project - USS Data, LLC: Cooperative Research and Development Final Report, CRADA Number CRD-09-367-04

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

Wilcox, S.

2013-08-01

Under this Agreement, NREL will work with Participant to improve concentrating solar power system performance characterizations. This work includes, but is not limited to, research and development of methods for acquiring renewable resource characterization information using site-specific measurements of solar radiation and meteorological conditions; collecting system performance data; and developing tools for improving the design, installation, operation, and maintenance of solar energy conversion systems. This work will be conducted at NREL and Participant facilities.

Solar Technology Validation Project - RES Americas: Cooperative Research and Development Final Report, CRADA Number CRD-09-367-11

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

Wilcox, S.

Under this Agreement, NREL will work with Participant to improve concentrating solar power system performance characterizations. This work includes, but is not limited to, research and development of methods for acquiring renewable resource characterization information using site-specific measurements of solar radiation and meteorological conditions; collecting system performance data; and developing tools for improving the design, installation, operation, and maintenance of solar energy conversion systems. This work will be conducted at NREL and Participant facilities.

Toward the Standardization of Biochar Analysis: The COST Action TD1107 Interlaboratory Comparison.

PubMed

Bachmann, Hans Jörg; Bucheli, Thomas D; Dieguez-Alonso, Alba; Fabbri, Daniele; Knicker, Heike; Schmidt, Hans-Peter; Ulbricht, Axel; Becker, Roland; Buscaroli, Alessandro; Buerge, Diane; Cross, Andrew; Dickinson, Dane; Enders, Akio; Esteves, Valdemar I; Evangelou, Michael W H; Fellet, Guido; Friedrich, Kevin; Gasco Guerrero, Gabriel; Glaser, Bruno; Hanke, Ulrich M; Hanley, Kelly; Hilber, Isabel; Kalderis, Dimitrios; Leifeld, Jens; Masek, Ondrej; Mumme, Jan; Carmona, Marina Paneque; Calvelo Pereira, Roberto; Rees, Frederic; Rombolà, Alessandro G; de la Rosa, José Maria; Sakrabani, Ruben; Sohi, Saran; Soja, Gerhard; Valagussa, Massimo; Verheijen, Frank; Zehetner, Franz

2016-01-20

Biochar produced by pyrolysis of organic residues is increasingly used for soil amendment and many other applications. However, analytical methods for its physical and chemical characterization are yet far from being specifically adapted, optimized, and standardized. Therefore, COST Action TD1107 conducted an interlaboratory comparison in which 22 laboratories from 12 countries analyzed three different types of biochar for 38 physical-chemical parameters (macro- and microelements, heavy metals, polycyclic aromatic hydrocarbons, pH, electrical conductivity, and specific surface area) with their preferential methods. The data were evaluated in detail using professional interlaboratory testing software. Whereas intralaboratory repeatability was generally good or at least acceptable, interlaboratory reproducibility was mostly not (20% < mean reproducibility standard deviation < 460%). This paper contributes to better comparability of biochar data published already and provides recommendations to improve and harmonize specific methods for biochar analysis in the future.

LC-MS/MS Identification of Species-Specific Muscle Peptides in Processed Animal Proteins.

PubMed

Marchis, Daniela; Altomare, Alessandra; Gili, Marilena; Ostorero, Federica; Khadjavi, Amina; Corona, Cristiano; Ru, Giuseppe; Cappelletti, Benedetta; Gianelli, Silvia; Amadeo, Francesca; Rumio, Cristiano; Carini, Marina; Aldini, Giancarlo; Casalone, Cristina

2017-12-06

An innovative analytical strategy has been applied to identify signature peptides able to distinguish among processed animal proteins (PAPs) derived from bovine, pig, fish, and milk products. Proteomics was first used to elucidate the proteome of each source. Starting from the identified proteins and using a funnel based approach, a set of abundant and well characterized peptides with suitable physical-chemical properties (signature peptides) and specific for each source was selected. An on-target LC-ESI-MS/MS method (MRM mode) was set up using standard peptides and was then applied to selectively identify the PAP source and also to distinguish proteins from bovine carcass and milk proteins. We believe that the method described meets the request of the European Commission which has developed a strategy for gradually lifting the "total ban" toward "species to species ban", therefore requiring official methods for species-specific discrimination in feed.

Lysozyme activity and nitroblue-tetrazolium reduction in leukaemic cells

PubMed Central

Catovsky, D.; Galton, D. A. G.

1973-01-01

The cytochemical methods for lysozyme and nitroblue-tetrazolium reduction have been used to study the blast cells of acute myeloid leukaemia. Both proved useful in characterizing the cases with predominant monocytic differentiation. The demonstration of lysozyme activity helped to define two main groups: (a) with predominantly lysozyme-negative cells (myeloblastic-promyelocytic), and (b) with considerable numbers of positive cells (monoblastic-monocytic). In addition this test was also of value in the differentiation of other leukaemic disorders. Reduction of nitroblue-tetrazolium was also a feature of monocytic differentiation. The combination of these two methods with those for myeloperoxidase and non-specific esterase activity contributes to the cytological characterization of acute myeloid leukaemia. Images PMID:4511938

Fuel Aging in Storage and Transportation (FAST): Accelerated Characterization and Performance Assessment of the Used Nuclear Fuel Storage System

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

McDeavitt, Sean

2016-08-02

This Integrated Research Project (IRP) was established to characterize key limiting phenomena related to the performance of used nuclear fuel (UNF) storage systems. This was an applied engineering project with a specific application in view (i.e., UNF dry storage). The completed tasks made use of a mixture of basic science and engineering methods. The overall objective was to create, or enable the creation of, predictive tools in the form of observation methods, phenomenological models, and databases that will enable the design, installation, and licensing of dry UNF storage systems that will be capable of containing UNF for extended period ofmore » time.« less

On the Use of Accelerated Test Methods for Characterization of Advanced Composite Materials

NASA Technical Reports Server (NTRS)

Gates, Thomas S.

2003-01-01

A rational approach to the problem of accelerated testing for material characterization of advanced polymer matrix composites is discussed. The experimental and analytical methods provided should be viewed as a set of tools useful in the screening of material systems for long-term engineering properties in aerospace applications. Consideration is given to long-term exposure in extreme environments that include elevated temperature, reduced temperature, moisture, oxygen, and mechanical load. Analytical formulations useful for predictive models that are based on the principles of time-based superposition are presented. The need for reproducible mechanisms, indicator properties, and real-time data are outlined as well as the methodologies for determining specific aging mechanisms.

Radioactive Waste Characterization Strategies; Comparisons Between AK/PK, Dose to Curie Modeling, Gamma Spectroscopy, and Laboratory Analysis Methods- 12194

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

Singledecker, Steven J.; Jones, Scotty W.; Dorries, Alison M.

2012-07-01

In the coming fiscal years of potentially declining budgets, Department of Energy facilities such as the Los Alamos National Laboratory (LANL) will be looking to reduce the cost of radioactive waste characterization, management, and disposal processes. At the core of this cost reduction process will be choosing the most cost effective, efficient, and accurate methods of radioactive waste characterization. Central to every radioactive waste management program is an effective and accurate waste characterization program. Choosing between methods can determine what is classified as low level radioactive waste (LLRW), transuranic waste (TRU), waste that can be disposed of under an Authorizedmore » Release Limit (ARL), industrial waste, and waste that can be disposed of in municipal landfills. The cost benefits of an accurate radioactive waste characterization program cannot be overstated. In addition, inaccurate radioactive waste characterization of radioactive waste can result in the incorrect classification of radioactive waste leading to higher disposal costs, Department of Transportation (DOT) violations, Notice of Violations (NOVs) from Federal and State regulatory agencies, waste rejection from disposal facilities, loss of operational capabilities, and loss of disposal options. Any one of these events could result in the program that mischaracterized the waste losing its ability to perform it primary operational mission. Generators that produce radioactive waste have four characterization strategies at their disposal: - Acceptable Knowledge/Process Knowledge (AK/PK); - Indirect characterization using a software application or other dose to curie methodologies; - Non-Destructive Analysis (NDA) tools such as gamma spectroscopy; - Direct sampling (e.g. grab samples or Surface Contaminated Object smears) and laboratory analytical; Each method has specific advantages and disadvantages. This paper will evaluate each method detailing those advantages and disadvantages including; - Cost benefit analysis (basic materials costs, overall program operations costs, man-hours per sample analyzed, etc.); - Radiation Exposure As Low As Reasonably Achievable (ALARA) program considerations; - Industrial Health and Safety risks; - Overall Analytical Confidence Level. The concepts in this paper apply to any organization with significant radioactive waste characterization and management activities working to within budget constraints and seeking to optimize their waste characterization strategies while reducing analytical costs. (authors)« less

Promoter isolation and characterization of GhAO-like1, a Gossypium hirsutum gene similar to multicopper oxidases that is highly expressed in reproductive organs.

PubMed

Lambret-Frotté, Julia; Artico, Sinara; Muniz Nardeli, Sarah; Fonseca, Fernando; Brilhante Oliveira-Neto, Osmundo; Grossi-de-Sá, Maria Fatima; Alves-Ferreira, Marcio

2016-01-01

Cotton is one of the most economically important cultivated crops. It is the major source of natural fiber for the textile industry and an important target for genetic modification for both biotic stress and herbicide tolerance. Therefore, the characterization of genes and regulatory regions that might be useful for genetic transformation is indispensable. The isolation and characterization of new regulatory regions is of great importance to drive transgene expression in genetically modified crops. One of the major drawbacks in cotton production is pest damage; therefore, the most promising, cost-effective, and sustainable method for pest control is the development of genetically resistant cotton lines. Considering this scenario, our group isolated and characterized the promoter region of a MCO (multicopper oxidase) from Gossypium hirsutum, named GhAO-like1 (ascorbate oxidase-like1). The quantitative expression, together with the in vivo characterization of the promoter region reveals that GhAO-like1 has a flower- and fruit-specific expression pattern. The GUS activity is mainly observed in stamens, as expected considering that the GhAO-like1 regulatory sequence is enriched in cis elements, which have been characterized as a target of reproductive tissue specific transcription factors. Both histological and quantitative analyses in Arabidopsis thaliana have confirmed flower (mainly in stamens) and fruit expression of GhAO-like1. In the present paper, we isolated and characterized both in silico and in vivo the promoter region of the GhAO-like1 gene. The regulatory region of GhAO-like1 might be useful to confer tissue-specific expression in genetically modified plants.

Assessment of multiple geophysical techniques for the characterization of municipal waste deposit sites

NASA Astrophysics Data System (ADS)

Gaël, Dumont; Tanguy, Robert; Nicolas, Marck; Frédéric, Nguyen

2017-10-01

In this study, we tested the ability of geophysical methods to characterize a large technical landfill installed in a former sand quarry. The geophysical surveys specifically aimed at delimitating the deposit site horizontal extension, at estimating its thickness and at characterizing the waste material composition (the moisture content in the present case). The site delimitation was conducted with electromagnetic (in-phase and out-of-phase) and magnetic (vertical gradient and total field) methods that clearly showed the transition between the waste deposit and the host formation. Regarding waste deposit thickness evaluation, electrical resistivity tomography appeared inefficient on this particularly thick deposit site. Thus, we propose a combination of horizontal to vertical noise spectral ratio (HVNSR) and multichannel analysis of the surface waves (MASW), which successfully determined the approximate waste deposit thickness in our test landfill. However, ERT appeared to be an appropriate tool to characterize the moisture content of the waste, which is of prior information for the organic waste biodegradation process. The global multi-scale and multi-method geophysical survey offers precious information for site rehabilitation studies, water content mitigation processes for enhanced biodegradation or landfill mining operation planning.

Mercapturic acids: recent advances in their determination by liquid chromatography/mass spectrometry and their use in toxicant metabolism studies and in occupational and environmental exposure studies

PubMed Central

Mathias, Patricia I.; B’Hymer, Clayton

2016-01-01

This review describes recent selected HPLC/MS methods for the determination of urinary mercapturates that are useful as non-invasive biomarkers in characterizing human exposure to electrophilic industrial chemicals in occupational and environmental studies. High performance liquid chromatography/mass spectrometry is a sensitive and specific method for analysis of small molecules found in biological fluids. In this review, recent selected mercapturate quantification methods are summarized and specific cases are presented. The biological formation of mercapturates is introduced and their use indicators of metabolic processing of reactive toxicants is discussed, as well as future trends and limitations in this area of research. PMID:26900903

Assessment of Technologies for the Space Shuttle External Tank Thermal Protection System and Recommendations for Technology Improvement - Part III: Material Property Characterization, Analysis, and Test Methods

NASA Technical Reports Server (NTRS)

Gates, Thomas S.; Johnson, Theodore F.; Whitley, Karen S.

2005-01-01

The objective of this report is to contribute to the independent assessment of the Space Shuttle External Tank Foam Material. This report specifically addresses material modeling, characterization testing, data reduction methods, and data pedigree. A brief description of the External Tank foam materials, locations, and standard failure modes is provided to develop suitable background information. A review of mechanics based analysis methods from the open literature is used to provide an assessment of the state-of-the-art in material modeling of closed cell foams. Further, this report assesses the existing material property database and investigates sources of material property variability. The report presents identified deficiencies in testing methods and procedures, recommendations for additional testing as required, identification of near-term improvements that should be pursued, and long-term capabilities or enhancements that should be developed.

A polyvalent harmonic coil testing method for small-aperture magnets

NASA Astrophysics Data System (ADS)

Arpaia, Pasquale; Buzio, Marco; Golluccio, Giancarlo; Walckiers, Louis

2012-08-01

A method to characterize permanent and fast-pulsed iron-dominated magnets with small apertures is presented. The harmonic coil measurement technique is enhanced specifically for small-aperture magnets by (1) in situ calibration, for facing search-coil production inaccuracy, (2) rotating the magnet around its axis, for correcting systematic effects, and (3) measuring magnetic fluxes by stationary coils at different angular positions for measuring fast pulsed magnets. This method allows a quadrupole magnet for particle accelerators to be characterized completely, by assessing multipole field components, magnetic axis position, and field direction. In this paper, initially the metrological problems arising from testing small-aperture magnets are highlighted. Then, the basic ideas of the proposed method and the architecture of the corresponding measurement system are illustrated. Finally, experimental validation results are shown for small-aperture permanent and fast-ramped quadrupole magnets for the new linear accelerator Linac4 at CERN (European Organization for Nuclear Research).

Computer-based objective quantitative assessment of pulmonary parenchyma via x-ray CT

NASA Astrophysics Data System (ADS)

Uppaluri, Renuka; McLennan, Geoffrey; Sonka, Milan; Hoffman, Eric A.

1998-07-01

This paper is a review of our recent studies using a texture- based tissue characterization method called the Adaptive Multiple Feature Method. This computerized method is automated and performs tissue classification based upon the training acquired on a set of representative examples. The AMFM has been applied to several different discrimination tasks including normal subjects, subjects with interstitial lung disease, smokers, asbestos-exposed subjects, and subjects with cystic fibrosis. The AMFM has also been applied to data acquired using different scanners and scanning protocols. The AMFM has shown to be successful and better than other existing techniques in discriminating the tissues under consideration. We demonstrate that the AMFM is considerably more sensitive and specific in characterizing the lung, especially in the presence of mixed pathology, as compared to more commonly used methods. Evidence is presented suggesting that the AMFM is highly sensitive to some of the earliest disease processes.

Characterization of Aspergillus section Nigri species populations in vineyard soil using droplet digital PCR

USDA-ARS?s Scientific Manuscript database

Identification of populations of Aspergillus section Nigri species in environmental samples using traditional methods is laborious and impractical for large numbers of samples. We developed species-specific primers and probes for quantitative droplet digital PCR (ddPCR) to improve sample throughput ...

Facile characterization of aptamer kinetic and equilibrium binding properties using surface plasmon resonance

PubMed Central

Chang, Andrew L.; McKeague, Maureen; Smolke, Christina D.

2015-01-01

Nucleic acid aptamers find widespread use as targeting and sensing agents in nature and biotechnology. Their ability to bind an extensive range of molecular targets, including small molecules, proteins, and ions, with high affinity and specificity enables their use in diverse diagnostic, therapeutic, imaging, and gene-regulatory applications. Here, we describe methods for characterizing aptamer kinetic and equilibrium binding properties using a surface plasmon resonance-based platform. This aptamer characterization platform is broadly useful for studying aptamer–ligand interactions, comparing aptamer properties, screening functional aptamers during in vitro selection processes, and prototyping aptamers for integration into nucleic acid devices. PMID:25432760

Characterization of Off-Road Diesel Emissions of Criteria Pollutants

DTIC Science & Technology

2008-10-01

Feasibility of soil dust source apportionment by the pyrolysis-gas chromatography/mass spectrometry method. J. Air Waste Manage. Assoc., 56(9):1230-1242...temperatures found in exhaust pipes. PM2.5 source profiles are important for speciated emission inventories and source apportionment , but few of these are...different between types. This method holds great potential for source apportionment , even in the absence of associating each pattern with a specific

"Spoligoriftyping," a dual-priming-oligonucleotide-based direct-hybridization assay for tuberculosis control with a multianalyte microbead-based hybridization system.

PubMed

Gomgnimbou, Michel Kiréopori; Abadia, Edgar; Zhang, Jian; Refrégier, Guislaine; Panaiotov, Stefan; Bachiyska, Elizabeta; Sola, Christophe

2012-10-01

We developed "spoligoriftyping," a 53-plex assay based on two preexisting methods, the spoligotyping and "rifoligotyping" assays, by combining them into a single assay. Spoligoriftyping allows simultaneous spoligotyping (i.e., clustered regularly interspaced short palindromic repeat [CRISPR]-based genotyping) and characterization of the main rifampin drug resistance mutations on the rpoB hot spot region in a few hours. This test partly uses the dual-priming-oligonucleotide (DPO) principle, which allows simultaneous efficient amplifications of rpoB and the CRISPR locus in the same sample. We tested this method on a set of 114 previously phenotypically and genotypically characterized multidrug-resistant (MDR) Mycobacterium tuberculosis or drug-susceptible M. tuberculosis DNA extracted from clinical isolates obtained from patients from Bulgaria, Nigeria, and Germany. We showed that our method is 100% concordant with rpoB sequencing results and 99.95% (3,911/3,913 spoligotype data points) correlated with classical spoligotyping results. The sensitivity and specificity of our assay were 99 and 100%, respectively, compared to those of phenotypic drug susceptibility testing. Such assays pave the way to the implementation of locally and specifically adapted methods of performing in a single tube both drug resistance mutation detection and genotyping in a few hours.

Current approaches used in epidemiologic studies to examine short-term multipollutant air pollution exposures.

PubMed

Davalos, Angel D; Luben, Thomas J; Herring, Amy H; Sacks, Jason D

2017-02-01

Air pollution epidemiology traditionally focuses on the relationship between individual air pollutants and health outcomes (e.g., mortality). To account for potential copollutant confounding, individual pollutant associations are often estimated by adjusting or controlling for other pollutants in the mixture. Recently, the need to characterize the relationship between health outcomes and the larger multipollutant mixture has been emphasized in an attempt to better protect public health and inform more sustainable air quality management decisions. New and innovative statistical methods to examine multipollutant exposures were identified through a broad literature search, with a specific focus on those statistical approaches currently used in epidemiologic studies of short-term exposures to criteria air pollutants (i.e., particulate matter, carbon monoxide, sulfur dioxide, nitrogen dioxide, and ozone). Five broad classes of statistical approaches were identified for examining associations between short-term multipollutant exposures and health outcomes, specifically additive main effects, effect measure modification, unsupervised dimension reduction, supervised dimension reduction, and nonparametric methods. These approaches are characterized including advantages and limitations in different epidemiologic scenarios. By highlighting the characteristics of various studies in which multipollutant statistical methods have been used, this review provides epidemiologists and biostatisticians with a resource to aid in the selection of the most optimal statistical method to use when examining multipollutant exposures. Published by Elsevier Inc.

Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution.

PubMed

Maeno, Takanori; Uzawa, Takanori; Kono, Izumi; Okano, Kazunori; Iino, Takanori; Fukita, Keisuke; Oshikawa, Yuki; Ogawa, Taro; Iwata, Osamu; Ito, Takuro; Suzuki, Kengo; Goda, Keisuke; Hosokawa, Yoichiroh

2018-05-29

Microalgae-based metabolic engineering has been proven effective for producing valuable substances such as food supplements, pharmaceutical drugs, biodegradable plastics, and biofuels in the past decade. The ability to accurately visualize and quantify intracellular metabolites in live microalgae is essential for efficient metabolic engineering, but remains a major challenge due to the lack of characterization methods. Here we demonstrate it by synthesizing fluorogenic peptide aptamers with specific binding affinity to a target metabolite and delivering them into live microalgae by femtosecond laser photoporation at single-cell resolution. As a proof-of-principle demonstration of our method, we use it to characterize Euglena gracilis, a photosynthetic unicellular motile microalgal species, which is capable of producing paramylon (a carbohydrate granule similar to starch). Specifically, we synthesize a peptide aptamer containing a paramylon-binding fluorescent probe, 7-nitrobenzofurazan, and introduce it into E. gracilis cells one-by-one by suppressing their mobility with mannitol and transiently perforating them with femtosecond laser pulses at 800 nm for photoporation. To demonstrate the method's practical utility in metabolic engineering, we perform spatially and temporally resolved fluorescence microscopy of single live photoporated E. gracilis cells under different culture conditions. Our method holds great promise for highly efficient microalgae-based metabolic engineering.

Isolation and chemical characterization of dissolved and colloidal organic matter

USGS Publications Warehouse

Aiken, G.; Leenheer, J.

1993-01-01

Commonly used techniques for the concentration and isolation of organic matter from water, such as preparative chromatography, ultrafiltration and reverse osmosis, and the methods used to analyze the organic matter obtained by these methods are reviewed. The development of methods to obtain organic matter that is associated with fractions of the dissolved organic carbon other than humic substances, such as organic bases, hydrophilic organic acids and colloidal organic matter are discussed. Methods specifically used to study dissolved organic nitrogen and dissolved organic phosphorous are also discussed. -from Authors

An original method to determine complex refractive index of liquids by spectroscopic ellipsometry and illustrated applications

NASA Astrophysics Data System (ADS)

Stchakovsky, M.; Battie, Y.; Naciri, A. En

2017-11-01

We present a method to characterize optical properties of liquids by spectroscopic ellipsometry. The experiments use a specific liquid cell that avoids disturbance of waves at air-liquid interface and allows the determination of the real and the imaginary part of the refractive index, with a sensitivity of the latter below 10-4. The method is illustrated by results obtained with a spectroscopic phase modulation ellipsometer on several liquids such as deionised water, microscope oil and protein solution. Comparisons of the method with standard techniques are given.

SEGMENTING CT PROSTATE IMAGES USING POPULATION AND PATIENT-SPECIFIC STATISTICS FOR RADIOTHERAPY.

PubMed

Feng, Qianjin; Foskey, Mark; Tang, Songyuan; Chen, Wufan; Shen, Dinggang

2009-08-07

This paper presents a new deformable model using both population and patient-specific statistics to segment the prostate from CT images. There are two novelties in the proposed method. First, a modified scale invariant feature transform (SIFT) local descriptor, which is more distinctive than general intensity and gradient features, is used to characterize the image features. Second, an online training approach is used to build the shape statistics for accurately capturing intra-patient variation, which is more important than inter-patient variation for prostate segmentation in clinical radiotherapy. Experimental results show that the proposed method is robust and accurate, suitable for clinical application.

SEGMENTING CT PROSTATE IMAGES USING POPULATION AND PATIENT-SPECIFIC STATISTICS FOR RADIOTHERAPY

PubMed Central

Feng, Qianjin; Foskey, Mark; Tang, Songyuan; Chen, Wufan; Shen, Dinggang

2010-01-01

This paper presents a new deformable model using both population and patient-specific statistics to segment the prostate from CT images. There are two novelties in the proposed method. First, a modified scale invariant feature transform (SIFT) local descriptor, which is more distinctive than general intensity and gradient features, is used to characterize the image features. Second, an online training approach is used to build the shape statistics for accurately capturing intra-patient variation, which is more important than inter-patient variation for prostate segmentation in clinical radiotherapy. Experimental results show that the proposed method is robust and accurate, suitable for clinical application. PMID:21197416

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Volume 4; Inherent Optical Properties: Instruments, Characterizations, Field Measurements and Data Analysis Protocols; Revised

NASA Technical Reports Server (NTRS)

Mueller, J. L. (Editor); Fargion, Giuletta S. (Editor); McClain, Charles R. (Editor); Pegau, Scott; Zaneveld, J. Ronald V.; Mitchell, B. Gregg; Kahru, Mati; Wieland, John; Stramska, Malgorzat

2003-01-01

This document stipulates protocols for measuring bio-optical and radiometric data for the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project activities and algorithm development. The document is organized into 6 separate volumes as Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 4. Volume I: Introduction, Background and Conventions; Volume II: Instrument Specifications, Characterization and Calibration; Volume III: Radiometric Measurements and Data Analysis Methods; Volume IV: Inherent Optical Properties: Instruments, Characterization, Field Measurements and Data Analysis Protocols; Volume V: Biogeochemical and Bio-Optical Measurements and Data Analysis Methods; Volume VI: Special Topics in Ocean Optics Protocols and Appendices. The earlier version of Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 3 (Mueller and Fargion 2002, Volumes 1 and 2) is entirely superseded by the six volumes of Revision 4 listed above.

Circulating Tumor Cells: Moving Biological Insights into Detection

PubMed Central

Chen, Lichan; Bode, Ann M; Dong, Zigang

2017-01-01

Circulating tumor cells (CTCs) have shown promising potential as liquid biopsies that facilitate early detection, prognosis, therapeutic target selection and monitoring treatment response. CTCs in most cancer patients are low in abundance and heterogeneous in morphological and phenotypic profiles, which complicate their enrichment and subsequent characterization. Several methodologies for CTC enrichment and characterization have been developed over the past few years. However, integrating recent advances in CTC biology into these methodologies and the selection of appropriate enrichment and characterization methods for specific applications are needed to improve the reliability of CTC biopsies. In this review, we summarize recent advances in the studies of CTC biology, including the mechanisms of their generation and their potential forms of existence in blood, as well as the current CTC enrichment technologies. We then critically examine the selection of methods for appropriately enriching CTCs for further investigation of their clinical applications. PMID:28819450

Characterization and stability studies of bioactive compounds and food matrices as evidence in support of health claims.

PubMed

González-Ferrero, Carolina; Sáiz-Abajo, María-José

2015-07-01

The characterization and stability evaluation of food and food constituents (chemical active ingredient/microorganism) for which nutrition or health claims want to be requested are essential for the success of an application to EFSA. This work reviews the requirements that must be fulfilled for a full characterization of the active substance, comprising origin, elaboration, or extraction method, and chemical/microbiological composition, using validated analytical methods. The review focuses not only on establishing the specifications of the final active ingredient or food but also on ensuring homogeneity between batches. In addition, the article discusses the methodologies and conditions of the stability studies that need to be performed on food and food constituents to verify that the relevant compounds--chemical and microbiological active ingredients--will get to the consumer in the intended state and concentration to accomplish the claimed health effect over shelf life.

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 4, Volume IV: Inherent Optical Properties: Instruments, Characterizations, Field Measurements and Data Analysis Protocols

NASA Technical Reports Server (NTRS)

Mueller, J. L.; Fargion, G. S.; McClain, C. R. (Editor); Pegau, S.; Zanefeld, J. R. V.; Mitchell, B. G.; Kahru, M.; Wieland, J.; Stramska, M.

2003-01-01

This document stipulates protocols for measuring bio-optical and radiometric data for the Sensor Intercomparision and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project activities and algorithm development. The document is organized into 6 separate volumes as Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 4. Volume I: Introduction, Background, and Conventions; Volume II: Instrument Specifications, Characterization and Calibration; Volume III: Radiometric Measurements and Data Analysis Methods; Volume IV: Inherent Optical Properties: Instruments, Characterization, Field Measurements and Data Analysis Protocols; Volume V: Biogeochemical and Bio-Optical Measurements and Data Analysis Methods; Volume VI: Special Topics in Ocean Optics Protocols and Appendices. The earlier version of Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 3 is entirely superseded by the six volumes of Revision 4 listed above.

A Novel Method for Characterizing Spacesuit Mobility Through Metabolic Cost

NASA Technical Reports Server (NTRS)

McFarland, Shane M.; Norcross, Jason R.

2014-01-01

Historically, spacesuit mobility has been characterized by directly measuring both range of motion and joint torque of individual anatomic joints. The work detailed herein aims to improve on this method, which is often prone to uncertainly, lack of repeatability, and a general lack of applicability to real-world functional tasks. Specifically, the goal of this work is to characterize suited mobility performance by directly measuring the metabolic performance of the occupant. Pilot testing was conducted in 2013, employing three subjects performing a range of functional tasks in two different suits prototypes, the Mark III and Z-1. Cursory analysis of the results shows the approach has merit, with consistent performance trends toward one suit over the other. Forward work includes the need to look at more subjects, a refined task set, and another suit in a different mass/mobility regime to validate the approach.

A simple methodology for characterization of germanium coaxial detectors by using Monte Carlo simulation and evolutionary algorithms.

PubMed

Guerra, J G; Rubiano, J G; Winter, G; Guerra, A G; Alonso, H; Arnedo, M A; Tejera, A; Gil, J M; Rodríguez, R; Martel, P; Bolivar, J P

2015-11-01

The determination in a sample of the activity concentration of a specific radionuclide by gamma spectrometry needs to know the full energy peak efficiency (FEPE) for the energy of interest. The difficulties related to the experimental calibration make it advisable to have alternative methods for FEPE determination, such as the simulation of the transport of photons in the crystal by the Monte Carlo method, which requires an accurate knowledge of the characteristics and geometry of the detector. The characterization process is mainly carried out by Canberra Industries Inc. using proprietary techniques and methodologies developed by that company. It is a costly procedure (due to shipping and to the cost of the process itself) and for some research laboratories an alternative in situ procedure can be very useful. The main goal of this paper is to find an alternative to this costly characterization process, by establishing a method for optimizing the parameters of characterizing the detector, through a computational procedure which could be reproduced at a standard research lab. This method consists in the determination of the detector geometric parameters by using Monte Carlo simulation in parallel with an optimization process, based on evolutionary algorithms, starting from a set of reference FEPEs determined experimentally or computationally. The proposed method has proven to be effective and simple to implement. It provides a set of characterization parameters which it has been successfully validated for different source-detector geometries, and also for a wide range of environmental samples and certified materials. Copyright © 2015 Elsevier Ltd. All rights reserved.

Nanoscale zero-valent metals: a review of synthesis, characterization, and applications to environmental remediation.

PubMed

Li, Lingyun; Hu, Jiwei; Shi, Xuedan; Fan, Mingyi; Luo, Jin; Wei, Xionghui

2016-09-01

Engineered nanoscale zero-valent metals (NZVMs) representing the forefront of technologies have been considered as promising materials for environmental remediation and antimicrobial effect, due to their high reducibility and strong adsorption capability. This review is focused on the methodology for synthesis of bare NZVMs, supported NZVMs, modified NZVMs, and bimetallic systems with both traditional and green methods. Recent studies have demonstrated that self-assembly methods can play an important role for obtaining ordered, controllable, and tunable NZVMs. In addition to common characterization methods, the state-of-the-art methods have been developed to obtain the properties of NZVMs (e.g., granularity, size distribution, specific surface area, shape, crystal form, and chemical bond) with the resolution down to subnanometer scale. These methods include spherical aberration corrected scanning transmission electron microscopy (Cs-corrected STEM), electron energy-loss spectroscopy (EELS), and near edge X-ray absorption fine structure (NEXAFS). A growing body of experimental data has proven that nanoscale zero-valent iron (NZVI) is highly effective and versatile. This article discusses the applications of NZVMs to treatment of heavy metals, halogenated organic compounds, polycyclic aromatic hydrocarbons, nutrients, radioelements, and microorganisms, using both ex situ and in situ methods. Furthermore, this paper briefly describes the ecotoxicological effects for NZVMs and the research prospects related to their synthesis, modification, characterization, and applications.

A Novel Method for Analyzing Microbially Affiliated Volatile Organic Compounds in Soil Environments

NASA Astrophysics Data System (ADS)

Ruhs, C. V.; McNeal, K. S.

2010-12-01

A concerted, international effort by citizens, governments, industries and educational systems is necessary to address the myriad environmental issues that face us today. The authors of this paper concentrate on soil environments and, specifically, the methods currently used to characterize them. The ability to efficiently and effectively monitor and characterize various soils is desired, allows for the study, supervision, and protection of natural and cultivated ecosystems, and may assist stakeholders in meeting governmentally-imposed environmental standards. This research addresses soil characterization by a comparison of four methods that emphasize a combination of microbial community and metabolic measures: BIOLOG, fatty acid methyl-ester analysis (FAME), descriptive physical and chemical analysis (moisture content, pH, carbon content, nutrient content, and grain size), and the novel soil-microbe volatile organic compound analysis (SMVOC) presented in this work. In order to achieve the method comparison, soils were collected from three climatic regions (Bahamas, Michigan, and Mississippi), with three samples taken from niche ecosystems found at each climatic region (a total of nine sites). Of interest to the authors is whether or not an investigation of microbial communities and the volatile organic compounds (VOCs) produced by microbial communities from nine separate soil ecosystems provides useful information about soil dynamics. In essence, is analysis of soil-derived VOCs using gas chromatography-mass spectrometry (GC-MS) an effective method for characterizing microbial communities and their metabolic activity of soils rapidly and accurately compared with the other three traditional characterization methods? Preliminary results suggest that VOCs in each of these locales differ with changes in soil types, soil moisture, and bacterial community. Each niche site shows distinct patterns in both VOCs and BIOLOG readings. Results will be presented to show the efficacy of the SMVOC approach and the statistical alignment of the VOC and community measures.

A high-resolution method for the localization of proanthocyanidins in plant tissues

PubMed Central

2011-01-01

Background Histochemical staining of plant tissues with 4-dimethylaminocinnamaldehyde (DMACA) or vanillin-HCl is widely used to characterize spatial patterns of proanthocyanidin accumulation in plant tissues. These methods are limited in their ability to allow high-resolution imaging of proanthocyanidin deposits. Results Tissue embedding techniques were used in combination with DMACA staining to analyze the accumulation of proanthocyanidins in Lotus corniculatus (L.) and Trifolium repens (L.) tissues. Embedding of plant tissues in LR White or paraffin matrices, with or without DMACA staining, preserved the physical integrity of the plant tissues, allowing high-resolution imaging that facilitated cell-specific localization of proanthocyanidins. A brown coloration was seen in proanthocyanidin-producing cells when plant tissues were embedded without DMACA staining and this was likely to have been due to non-enzymatic oxidation of proanthocyanidins and the formation of colored semiquinones and quinones. Conclusions This paper presents a simple, high-resolution method for analysis of proanthocyanidin accumulation in organs, tissues and cells of two plant species with different patterns of proanthocyanidin accumulation, namely Lotus corniculatus (birdsfoot trefoil) and Trifolium repens (white clover). This technique was used to characterize cell type-specific patterns of proanthocyanidin accumulation in white clover flowers at different stages of development. PMID:21595992

Quantifying stream nutrient uptake from ambient to saturation with instantaneous tracer additions

NASA Astrophysics Data System (ADS)

Covino, T. P.; McGlynn, B. L.; McNamara, R.

2009-12-01

Stream nutrient tracer additions and spiraling metrics are frequently used to quantify stream ecosystem behavior. However, standard approaches limit our understanding of aquatic biogeochemistry. Specifically, the relationship between in-stream nutrient concentration and stream nutrient spiraling has not been characterized. The standard constant rate (steady-state) approach to stream spiraling parameter estimation, either through elevating nutrient concentration or adding isotopically labeled tracers (e.g. 15N), provides little information regarding the stream kinetic curve that represents the uptake-concentration relationship analogous to the Michaelis-Menten curve. These standard approaches provide single or a few data points and often focus on estimating ambient uptake under the conditions at the time of the experiment. Here we outline and demonstrate a new method using instantaneous nutrient additions and dynamic analyses of breakthrough curve (BTC) data to characterize the full relationship between spiraling metrics and nutrient concentration. We compare the results from these dynamic analyses to BTC-integrated, and standard steady-state approaches. Our results indicate good agreement between these three approaches but we highlight the advantages of our dynamic method. Specifically, our new dynamic method provides a cost-effective and efficient approach to: 1) characterize full concentration-spiraling metric curves; 2) estimate ambient spiraling metrics; 3) estimate Michaelis-Menten parameters maximum uptake (Umax) and the half-saturation constant (Km) from developed uptake-concentration kinetic curves, and; 4) measure dynamic nutrient spiraling in larger rivers where steady-state approaches are impractical.

Using Atomic Force Microscopy to Characterize the Conformational Properties of Proteins and Protein-DNA Complexes That Carry Out DNA Repair.

PubMed

LeBlanc, Sharonda; Wilkins, Hunter; Li, Zimeng; Kaur, Parminder; Wang, Hong; Erie, Dorothy A

2017-01-01

Atomic force microscopy (AFM) is a scanning probe technique that allows visualization of single biomolecules and complexes deposited on a surface with nanometer resolution. AFM is a powerful tool for characterizing protein-protein and protein-DNA interactions. It can be used to capture snapshots of protein-DNA solution dynamics, which in turn, enables the characterization of the conformational properties of transient protein-protein and protein-DNA interactions. With AFM, it is possible to determine the stoichiometries and binding affinities of protein-protein and protein-DNA associations, the specificity of proteins binding to specific sites on DNA, and the conformations of the complexes. We describe methods to prepare and deposit samples, including surface treatments for optimal depositions, and how to quantitatively analyze images. We also discuss a new electrostatic force imaging technique called DREEM, which allows the visualization of the path of DNA within proteins in protein-DNA complexes. Collectively, these methods facilitate the development of comprehensive models of DNA repair and provide a broader understanding of all protein-protein and protein-nucleic acid interactions. The structural details gleaned from analysis of AFM images coupled with biochemistry provide vital information toward establishing the structure-function relationships that govern DNA repair processes. © 2017 Elsevier Inc. All rights reserved.

Molecular-Level Study of the Effect of Prior Axial Compression/Torsion on the Axial-Tensile Strength of PPTA Fibers

DTIC Science & Technology

2013-07-16

Twaron, etc., which are characterized by high specific strength and high specific stiffness. Fibers of this type are often referred to as ‘‘ballistic... high level of penetration resistance against large kinetic energy projectiles, such as bullets, detonated-mine-induced soil ejecta, improvised...increasingly being designed and developed through an extensive use of computer-aided engineering ( CAE ) methods and tools. The utility of these

Axial-Compressive Behavior, Including Kink-Band Formation and Propagation, of Single p-Phenylene Terephthalamide (PPTA) Fibers

DTIC Science & Technology

2013-01-01

material models to describe the behavior of fibers and structures under high -rate loading conditions. With the utility of the CAE methods and tools largely...phenylene terephthalamide (PPTA), available commercially as Kevlar, Twaron, Technora, and so forth, are characterized by high specific axial stiffness...and high specific tensile strength. These fibers are often referred to as “ballistic fibers” since they are commonly used in different ballistic- and

Defining the Developmental Parameters of Temper Loss in Early Childhood: Implications for Developmental Psychopathology

ERIC Educational Resources Information Center

Wakschlag, Lauren S.; Choi, Seung W.; Carter, Alice S.; Hullsiek, Heide; Burns, James; McCarthy, Kimberly; Leibenluft, Ellen; Briggs-Gowan, Margaret J.

2012-01-01

Background: Temper modulation problems are both a hallmark of early childhood and a common mental health concern. Thus, characterizing specific behavioral manifestations of temper loss along a dimension from normative misbehaviors to clinically significant problems is an important step toward identifying clinical thresholds. Methods:…

Plotting landscape perspectives of clearcut units.

Treesearch

Roger H. Twito

1978-01-01

The aesthetics of clearcut units potentially visible from a distance can be improved by judicious design. Screening clearcuts from view or shaping them to characterize natural openings are current methods used for this purpose. Perspective plots illustrating how proposed clearcut units will look from specific off-site viewing points provide an...

Isolation by ion-exchange methods. In Sarker S.D. (ed) Natural Products Isolation, 3rd edition

USDA-ARS?s Scientific Manuscript database

The primary goal of many natural products chemists is to extract, isolate, and characterize specific analytes from complex plant, animal, microbial, and food matrices. To achieve this goal, they rely considerably on highly sophisticated and highly hyphenated modern instrumentation. Yet, the vast maj...

TRACE-LEVEL MEASUREMENT OF COMPLEX COMBUSTION EFFLUENTS AND RESIDUES USING MULTIDIMENSIONAL GAS CHROMATOGRAPHY-MASS SPECTROMETRY (MDGC-MS). (R828190)

EPA Science Inventory

The identification and quantitation of non-method-specific target analytes have greater importance with respect to EPA's current combustion strategy. The risk associated with combustion process emissions must now be characterized. EPA has recently released draft guidance on pr...

Method and apparatus for ion mobility spectrometry with alignment of dipole direction (IMS-ADD)

DOEpatents

Shvartsburg, Alexandre A [Richland, WA; Tang, Keqi [Richland, WA; Smith, Richard D [Richland, WA

2007-01-30

Techniques and instrumentation are described for analyses of substances, including complex samples/mixtures that require separation prior to characterization of individual components. A method is disclosed for separation of ion mixtures and identification of ions, including protein and other macromolecular ions and their different structural isomers. Analyte ions are not free to rotate during the separation, but are substantially oriented with respect to the drift direction. Alignment is achieved by applying, at a particular angle to the drift field, a much stronger alternating electric field that "locks" the ion dipoles with moments exceeding a certain value. That value depends on the buffer gas composition, pressure, and temperature, but may be as low as .about.3 Debye under certain conditions. The presently disclosed method measures the direction-specific cross-sections that provide the structural information complementing that obtained from known methods, and, when coupled to those methods, increases the total peak capacity and specificity of gas-phase separations. Simultaneous 2-D separations by direction-specific cross sections along and orthogonally to the ion dipole direction are also possible.

SPECIFICITY OF IMPROVED METHODS FOR MYCOBACTIN BIOASSAY BY ARTHROBACTER TERREGENS

PubMed Central

Antoine, Alan D.; Morrison, Norman E.; Hanks, John H.

1964-01-01

Antoine, Alan D. (Johns Hopkins University-Leonard Wood Memorial Leprosy Research Laboratory, Baltimore, Md.), Norman E. Morrison, and John H. Hanks. Specificity of improved methods for mycobactin bioassay by Arthrobacter terregens. J. Bacteriol. 88:1672–1677. 1964.—Arthrobacter terregens was used to assay mycobactin, a growth factor for Mycobacterium paratuberculosis. Improved techniques permit the assay of mycobactin within 3 to 4 days by agarplate or liquid-medium methods. For the agarplate method, Arthrobacter terregens gave linear increases in zonal growth at mycobactin concentrations of 0.07 to 0.30 μg per spot; for the liquid-medium method, linear increases in turbidimetric growth occurred at 0.05 to 0.27 μg/ml. Specificity studies show that the mycobactin hydrolytic products, cobactin and mycobactic acid, function as growth stimulators, but the high concentrations required would produce only minimal interference in mycobactin assays. Furthermore, the response to mycobactic acid is characterized by a delayed response of 3 days. Various synthetic hydroxylamine-containing compounds and metalchelating agents cannot replace the biological activity of mycobactin. Diacetylmycobactin is 7.4 times more effective than mycobactin as a growth stimulator. PMID:14240956

Temperature sensitive surfaces and methods of making same

DOEpatents

Liang, Liang [Richland, WA; Rieke, Peter C [Pasco, WA; Alford, Kentin L [Pasco, WA

2002-09-10

Poly-n-isopropylacrylamide surface coatings demonstrate the useful property of being able to switch charateristics depending upon temperature. More specifically, these coatings switch from being hydrophilic at low temperature to hydrophobic at high temperature. Research has been conducted for many years to better characterize and control the properties of temperature sensitive coatings. The present invention provides novel temperature sensitive coatings on articles and novel methods of making temperature sensitive coatings that are disposed on the surfaces of various articles. These novel coatings contain the reaction products of n-isopropylacrylamide and are characterized by their properties such as advancing contact angles. Numerous other characteristics such as coating thickness, surface roughness, and hydrophilic-to-hydrophobic transition temperatures are also described. The present invention includes articles having temperature-sensitve coatings with improved properties as well as improved methods for forming temperature sensitive coatings.

Interference of avian guano in analyses of fuel-contaminated soils

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

James, D.E.; Johnson, T.E.; Kreamer, D.K.

1996-01-01

Site characterization on Johnston Island, Johnston Atoll, Pacific Ocean, has yielded preliminary data that seabird guano can be an interference in three common petroleum hydrocarbon quantification methods. Volatiles from seabird guano were measured on a hydrocarbon-specific handheld vapor meter (catalytic detector) in concentrations as high as 256 ppm by volume total hydrocarbon. Analysis of guano solids produced measurable concentrations of total petroleum hydrocarbon (TPH) as diesel using both an immunoassay test and the EPA 8015 Modified Method. The testing was conducted on one surface sample of guano collected from a seabird roosting and nesting area. Source species were not identified.more » Positive hydrocarbon test results for guano raise concerns regarding the effectiveness of standard methods of petroleum-contaminated site characterization for Johnston island, other Pacific islands, and coastal areas with historic or contemporary seabird populations.« less

Influence of Humic Acids Extracted from Peat by Different Methods on Functional Activity of Macrophages in Vitro.

PubMed

Trofimova, E S; Zykova, M V; Ligacheva, A A; Sherstoboev, E Y; Zhdanov, V V; Belousov, M V; Yusubov, M S; Krivoshchekov, S V; Danilets, M G; Dygai, A M

2017-04-01

We studied activation of macrophages with humic acids extracted from peat of large deposits in the Tomsk region by two extraction methods: by hydroxide or sodium pyrophosphate. Humic acid of lowland peat types containing large amounts of aromatic carbon, phenolic and alcohol groups, carbohydrate residues and ethers, irrespectively of the extraction methods contained LPS admixture that probably determines their activating properties. Humic acid of upland peat types characterized by high content of carbonyl, carboxyl, and ester groups enhance NO production and reduce arginase expression, but these effects were minimized when sodium hydroxide was used as an extraction solvent. Pyrophosphate samples of the upland peat types were characterized by aromaticity and diversity of functional groups and have a significant advantage because of they induce specific endotoxin-independent stimulating action on antigen presenting cells.

A comparative in silico linear B-cell epitope prediction and characterization for South American and African Trypanosoma vivax strains.

PubMed

Guedes, Rafael Lucas Muniz; Rodrigues, Carla Monadeli Filgueira; Coatnoan, Nicolas; Cosson, Alain; Cadioli, Fabiano Antonio; Garcia, Herakles Antonio; Gerber, Alexandra Lehmkuhl; Machado, Rosangela Zacarias; Minoprio, Paola Marcella Camargo; Teixeira, Marta Maria Geraldes; de Vasconcelos, Ana Tereza Ribeiro

2018-02-27

Trypanosoma vivax is a parasite widespread across Africa and South America. Immunological methods using recombinant antigens have been developed aiming at specific and sensitive detection of infections caused by T. vivax. Here, we sequenced for the first time the transcriptome of a virulent T. vivax strain (Lins), isolated from an outbreak of severe disease in South America (Brazil) and performed a computational integrated analysis of genome, transcriptome and in silico predictions to identify and characterize putative linear B-cell epitopes from African and South American T. vivax. A total of 2278, 3936 and 4062 linear B-cell epitopes were respectively characterized for the transcriptomes of T. vivax LIEM-176 (Venezuela), T. vivax IL1392 (Nigeria) and T. vivax Lins (Brazil) and 4684 for the genome of T. vivax Y486 (Nigeria). The results presented are a valuable theoretical source that may pave the way for highly sensitive and specific diagnostic tools. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Scaling Linguistic Characterization of Precipitation Variability

NASA Astrophysics Data System (ADS)

Primo, C.; Gutierrez, J. M.

2003-04-01

Rainfall variability is influenced by changes in the aggregation of daily rainfall. This problem is of great importance for hydrological, agricultural and ecological applications. Rainfall averages, or accumulations, are widely used as standard climatic parameters. However different aggregation schemes may lead to the same average or accumulated values. In this paper we present a fractal method to characterize different aggregation schemes. The method provides scaling exponents characterizing weekly or monthly rainfall patterns for a given station. To this aim, we establish an analogy with linguistic analysis, considering precipitation as a discrete variable (e.g., rain, no rain). Each weekly, or monthly, symbolic precipitation sequence of observed precipitation is then considered as a "word" (in this case, a binary word) which defines a specific weekly rainfall pattern. Thus, each site defines a "language" characterized by the words observed in that site during a period representative of the climatology. Then, the more variable the observed weekly precipitation sequences, the more complex the obtained language. To characterize these languages, we first applied the Zipf's method obtaining scaling histograms of rank ordered frequencies. However, to obtain significant exponents, the scaling must be maintained some orders of magnitude, requiring long sequences of daily precipitation which are not available at particular stations. Thus this analysis is not suitable for applications involving particular stations (such as regionalization). Then, we introduce an alternative fractal method applicable to data from local stations. The so-called Chaos-Game method uses Iterated Function Systems (IFS) for graphically representing rainfall languages, in a way that complex languages define complex graphical patterns. The box-counting dimension and the entropy of the resulting patterns are used as linguistic parameters to quantitatively characterize the complexity of the patterns. We illustrate the high climatological discrimination power of the linguistic parameters in the Iberian peninsula, when compared with other standard techniques (such as seasonal mean accumulated precipitation). As an example, standard and linguistic parameters are used as inputs for a clustering regionalization method, comparing the resulting clusters.

Can we develop pathology-specific MRI contrast for "MR-negative" epilepsy?

PubMed

Feindel, Kirk W

2013-05-01

Recent improvements in magnetic resonance imaging (MRI) hardware, software, and analysis routines are helping to put cases of "MR-negative" epilepsy on the decline. However, most standard-of-care MRI relies on careful manipulation and presentation of T1, T2, and diffusion-weighted contrast, which characterize the behavior of water in "bulk" tissue rather than providing pathology-specific contrast. Research efforts in MR physics continue to identify and develop novel theory, and methods such as diffusional kurtosis imaging (DKI) and temporal diffusion spectroscopy that can better characterize tissue substructure, and chemical exchange saturation transfer (CEST) that can target underlying biochemical processes. The potential role of each technique in targeting pathologies implicated in "MR-negative" epilepsy is outlined herein. Wiley Periodicals, Inc. © 2013 International League Against Epilepsy.

An overview of field-specific designs of microbial EOR

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

Robertson, E.P.; Bala, G.A.; Fox, S.L.

1995-12-31

The selection and design of an MEOR process for application in a specific field involves geological, reservoir, and biological characterization. Microbially mediated oil recovery mechanisms (bigenic gas, biopolymers, and biosurfactants) are defined by the types of microorganisms used. The engineering and biological character of a given reservoir must be understood to correctly select a microbial system to enhance oil recovery. This paper discusses the methods used to evaluate three fields with distinct characteristics and production problems for the applicability of MEOR would not be applicable in two of the three fields considered. The development of a microbial oil recovery processmore » for the third field appeared promising. Development of a bacterial consortium capable of producing the desired metabolites was initiated, and field isolates were characterized.« less

EBIC Characterization and Hydrogen Passivation in Silicon Sheet

NASA Technical Reports Server (NTRS)

Hanoka, J. I.

1985-01-01

As a general qualitative tool, the electron beam induced current (EBIC) method can be very useful in imaging recombination in silicon sheet used for solar cells. Work using EBIC on EFG silicon ribbon is described. In particular, some efforts at making the technique more quantitative and hence more useful, some limitations of the method, and finally specific application to hydrogen passivation is treated. Some brief remarks are made regarding the technique itself.

Rapid functional screening of Streptomyces coelicolor regulators by use of a pH indicator and application to the MarR-like regulator AbsC.

PubMed

Yang, Yung-Hun; Song, Eunjung; Lee, Bo-Rahm; Kim, Eun-jung; Park, Sung-Hee; Kim, Yun-Gon; Lee, Chang-Soo; Kim, Byung-Gee

2010-06-01

To elucidate the function of an unknown regulator in Streptomyces, differences in phenotype and antibiotic production between a deletion mutant and a wild-type strain (WT) were compared. These differences are easily hidden by complex media. To determine the specific nutrient conditions that reveal such differences, we used a multiwell method containing different nutrients along with bromothymol blue. We found several nutrients that provide key information on characterization conditions. By comparing the growth of wild-type and mutant strains on screened nutrients, we were able to measure growth, organic acid production, and antibiotic production for the elucidation of regulator function. As a result of this method, a member of the MarR-like regulator family, SCO5405 (AbsC), was newly characterized to control pyruvate dehydrogenase in Streptomyces coelicolor. Deletion of SCO5405 increased the pH of the culture broth due to decreased production of organic acids such as pyruvate and alpha-ketoglutarate and increased extracellular actinorhodin (ACT) production in minimal medium containing glucose and alanine (MMGA). This method could therefore be a high-throughput method for the characterization of unknown regulators.

Flanking sequence determination and specific PCR identification of transgenic wheat B102-1-2.

PubMed

Cao, Jijuan; Xu, Junyi; Zhao, Tongtong; Cao, Dongmei; Huang, Xin; Zhang, Piqiao; Luan, Fengxia

2014-01-01

The exogenous fragment sequence and flanking sequence between the exogenous fragment and recombinant chromosome of transgenic wheat B102-1-2 were successfully acquired using genome walking technology. The newly acquired exogenous fragment encoded the full-length sequence of transformed genes with transformed plasmid and corresponding functional genes including ubi, vector pBANF-bar, vector pUbiGUSPlus, vector HSP, reporter vector pUbiGUSPlus, promoter ubiquitin, and coli DH1. A specific polymerase chain reaction (PCR) identification method for transgenic wheat B102-1-2 was established on the basis of designed primers according to flanking sequence. This established specific PCR strategy was validated by using transgenic wheat, transgenic corn, transgenic soybean, transgenic rice, and non-transgenic wheat. A specifically amplified target band was observed only in transgenic wheat B102-1-2. Therefore, this method is characterized by high specificity, high reproducibility, rapid identification, and excellent accuracy for the identification of transgenic wheat B102-1-2.

Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes

PubMed Central

Greenough, Lucia; Schermerhorn, Kelly M.; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E.; Gardner, Andrew F.

2016-01-01

Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3′-5′ exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. PMID:26365239

Characterizing human activity induced impulse and slip-pulse excitations through structural vibration

NASA Astrophysics Data System (ADS)

Pan, Shijia; Mirshekari, Mostafa; Fagert, Jonathon; Ramirez, Ceferino Gabriel; Chung, Albert Jin; Hu, Chih Chi; Shen, John Paul; Zhang, Pei; Noh, Hae Young

2018-02-01

Many human activities induce excitations on ambient structures with various objects, causing the structures to vibrate. Accurate vibration excitation source detection and characterization enable human activity information inference, hence allowing human activity monitoring for various smart building applications. By utilizing structural vibrations, we can achieve sparse and non-intrusive sensing, unlike pressure- and vision-based methods. Many approaches have been presented on vibration-based source characterization, and they often either focus on one excitation type or have limited performance due to the dispersion and attenuation effects of the structures. In this paper, we present our method to characterize two main types of excitations induced by human activities (impulse and slip-pulse) on multiple structures. By understanding the physical properties of waves and their propagation, the system can achieve accurate excitation tracking on different structures without large-scale labeled training data. Specifically, our algorithm takes properties of surface waves generated by impulse and of body waves generated by slip-pulse into account to handle the dispersion and attenuation effects when different types of excitations happen on various structures. We then evaluate the algorithm through multiple scenarios. Our method achieves up to a six times improvement in impulse localization accuracy and a three times improvement in slip-pulse trajectory length estimation compared to existing methods that do not take wave properties into account.

Facile synthesis of nickel-based metal organic framework [Ni3(HCOO)6] by microwave method and application for supercapacitor

NASA Astrophysics Data System (ADS)

Luo, Jujie; Yang, Xing; Wang, Shumin; Bi, Yuhong; Nautiyal, Amit; Zhang, Xinyu

The metal organic framework (MOF) [Ni3(HCOO)6] was synthesized via the simple and fast microwave method, and the effect of irradiation power on crystallinity of synthesized Ni-based MOF was studied. The samples were characterized by using Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The synthesized Ni-based MOF was electrochemically characterized by using galvanostatic charge-discharge (GCD), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) techniques. The synthesized MOF showed the highest specific capacitance of 1196.2F/g at 1A/g with excellent cyclability (86.04% capacitance retention after 2,000 cycles), thereby demonstrating its potential application in supercapacitors.

Investigation of specific interactions between T7 promoter and T7 RNA polymerase by force spectroscopy using atomic force microscope.

PubMed

Zhang, Xiaojuan; Yao, Zhixuan; Duan, Yanting; Zhang, Xiaomei; Shi, Jinsong; Xu, Zhenghong

2018-01-11

The specific recognition and binding of promoter and RNA polymerase is the first step of transcription initiation in bacteria and largely determines transcription activity. Therefore, direct analysis of the interaction between promoter and RNA polymerase in vitro may be a new strategy for promoter characterization, to avoid interference due to the cell's biophysical condition and other regulatory elements. In the present study, the specific interaction between T7 promoter and T7 RNA polymerase was studied as a model system using force spectroscopy based on atomic force microscope (AFM). The specific interaction between T7 promoter and T7 RNA polymerase was verified by control experiments, and the rupture force in this system was measured as 307.2 ± 6.7 pN. The binding between T7 promoter mutants with various promoter activities and T7 RNA polymerase was analyzed. Interaction information including rupture force, rupture distance and binding percentage were obtained in vitro , and reporter gene expression regulated by these promoters was also measured according to a traditional promoter activity characterization method in vivo Using correlation analysis, it was found that the promoter strength characterized by reporter gene expression was closely correlated with rupture force and the binding percentage by force spectroscopy. These results indicated that the analysis of the interaction between promoter and RNA polymerase using AFM-based force spectroscopy was an effective and valid approach for the quantitative characterization of promoters. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Model-independent Exoplanet Transit Spectroscopy

NASA Astrophysics Data System (ADS)

Aronson, Erik; Piskunov, Nikolai

2018-05-01

We propose a new data analysis method for obtaining transmission spectra of exoplanet atmospheres and brightness variation across the stellar disk from transit observations. The new method is capable of recovering exoplanet atmosphere absorption spectra and stellar specific intensities without relying on theoretical models of stars and planets. We simultaneously fit both stellar specific intensity and planetary radius directly to transit light curves. This allows stellar models to be removed from the data analysis. Furthermore, we use a data quality weighted filtering technique to achieve an optimal trade-off between spectral resolution and reconstruction fidelity homogenizing the signal-to-noise ratio across the wavelength range. Such an approach is more efficient than conventional data binning onto a low-resolution wavelength grid. We demonstrate that our analysis is capable of reproducing results achieved by using an explicit quadratic limb-darkening equation and that the filtering technique helps eliminate spurious spectral features in regions with strong telluric absorption. The method is applied to the VLT FORS2 observations of the exoplanets GJ 1214 b and WASP-49 b, and our results are in agreement with previous studies. Comparisons between obtained stellar specific intensity and numerical models indicates that the method is capable of accurately reconstructing the specific intensity. The proposed method enables more robust characterization of exoplanetary atmospheres by separating derivation of planetary transmission and stellar specific intensity spectra (that is model-independent) from chemical and physical interpretation.

Characterization and optimization of polycrystalline Si70%Ge30% for surface micromachined thermopiles in human body applications

NASA Astrophysics Data System (ADS)

Wang, Ziyang; Fiorini, Paolo; Leonov, Vladimir; Van Hoof, Chris

2009-09-01

This paper presents the material characterization methods, characterization results and the optimization scheme for polycrystalline Si70%Ge30% (poly-SiGe) from the perspective of its application in a surface micromachined thermopile. Due to its comparative advantages, such as lower thermal conductivity and ease of processing, over other materials, poly-SiGe is chosen to fabricate a surface micromachined thermopile and eventually a wearable thermoelectric generator (TEG) to be used on a human body. To enable optimal design of advanced thermocouple microstructures, poly-SiGe sample materials prepared by two different techniques, namely low-pressure chemical vapor deposition (LPCVD) with in situ doping and rapid thermal chemical vapor deposition (RTCVD) with ion implantation, have been characterized. Relevant material properties, including electrical resistivity, Seebeck coefficient, thermal conductivity and specific contact resistance, have been reported. For the determination of thermal conductivity, a novel surface-micromachined test structure based on the Seebeck effect is designed, fabricated and measured. Compared to the traditional test structures, it is more advantageous for sample materials with a relatively large Seebeck coefficient, such as poly-SiGe. Based on the characterization results, a further optimization scheme is suggested to allow independent respective optimization of the figure of merit and the specific contact resistance.

Evaluation of smallpox vaccines using variola neutralization.

PubMed

Damon, Inger K; Davidson, Whitni B; Hughes, Christine M; Olson, Victoria A; Smith, Scott K; Holman, Robert C; Frey, Sharon E; Newman, Frances; Belshe, Robert B; Yan, Lihan; Karem, Kevin

2009-08-01

The search for a 'third'-generation smallpox vaccine has resulted in the development and characterization of several vaccine candidates. A significant barrier to acceptance is the absence of challenge models showing induction of correlates of protective immunity against variola virus. In this light, virus neutralization provides one of few experimental methods to show specific 'in vitro' activity of vaccines against variola virus. Here, we provide characterization of the ability of a modified vaccinia virus Ankara vaccine to induce variola virus-neutralizing antibodies, and we provide comparison with the neutralization elicited by standard Dryvax vaccination.

Demonstrating Interactions of Transcription Factors with DNA by Electrophoretic Mobility Shift Assay.

PubMed

Yousaf, Nasim; Gould, David

2017-01-01

Confirming the binding of a transcription factor with a particular DNA sequence may be important in characterizing interactions with a synthetic promoter. Electrophoretic mobility shift assay is a powerful approach to demonstrate the specific DNA sequence that is bound by a transcription factor and also to confirm the specific transcription factor involved in the interaction. In this chapter we describe a method we have successfully used to demonstrate interactions of endogenous transcription factors with sequences derived from endogenous and synthetic promoters.

Matrix Assisted Laser Desorption Ionization Mass Spectrometric Analysis of Bacillus anthracis: From Fingerprint Analysis of the Bacterium to Quantification of its Toxins in Clinical Samples

NASA Astrophysics Data System (ADS)

Woolfitt, Adrian R.; Boyer, Anne E.; Quinn, Conrad P.; Hoffmaster, Alex R.; Kozel, Thomas R.; de, Barun K.; Gallegos, Maribel; Moura, Hercules; Pirkle, James L.; Barr, John R.

A range of mass spectrometry-based techniques have been used to identify, characterize and differentiate Bacillus anthracis, both in culture for forensic applications and for diagnosis during infection. This range of techniques could usefully be considered to exist as a continuum, based on the degrees of specificity involved. We show two examples here, a whole-organism fingerprinting method and a high-specificity assay for one unique protein, anthrax lethal factor.

[The structure of the glycerophosphate-containing O-specific polysaccharide from Escherichia coli 0130].

PubMed

Perepelov, A V; Lu, B; Sebchenkova, S N; Shevelev, S D; Wang, V; Shashkov, A S; Feng, L; Wang, L; Knirel', Iu A

2007-01-01

A phosphorylated O-specific polysaccharide was obtained by mild acidic degradation of the lipopolysaccharide from the intestinal bacterium Escherichia coli 0130 and characterized by the methods of chemical analysis, including dephosphorylation, and 1H and 13C NMR spectroscopy. The polysaccharide was shown to be composed of branched tetrasaccharide repeating units containing two N-acetyl-D-galactosamine residues, D-galactose, D-glucose, and glycerophosphate residues (one of each). The polysaccharide has the following structure, which is unique among the known bacterial polysaccharides.

Cluster model studies of anion and molecular specificities via electrospray ionization photoelectron spectroscopy

DOE PAGES

Wang, Xue -Bin

2017-01-06

Ion specificity, a widely observed macroscopic phenomenon in condensed phases and at interfaces, is essentially a fundamental chemical physical issue. We have been investigating such effects using cluster models in an “atom-by-atom” and “molecule-by-molecule” fashion not possible with condensed-phase methods. We use electrospray ionization (ESI) to generate molecular and ionic clusters to simulate key molecular entities involved in local binding regions, and characterize them employing negative ion photoelectron spectroscopy (NIPES). Inter- and intramolecular interactions and binding configurations are directly obtained as functions of cluster size and composition, providing insightful molecular-level description and characterization over the local active sites that playmore » crucial roles in determining solution chemistry and condensed phase phenomena. Finally, the topics covered in this article are relevant to a wide scope of research fields ranging from ion specific effects in electrolyte solutions, ion selectivity/recognition in normal functioning of life, to molecular specificity in aerosol particle formation, as well as in rational material design and synthesis.« less

Characterization of Aspergillus flavus strains from Brazilian Brazil nuts and cashew by RAPD and ribosomal DNA analysis.

PubMed

Midorikawa, G E O; Pinheiro, M R R; Vidigal, B S; Arruda, M C; Costa, F F; Pappas, G J; Ribeiro, S G; Freire, F; Miller, R N G

2008-07-01

The aim of this study was to determine the genetic variability in Aspergillus flavus populations from Brazil nut and cashew and develop a polymerase chain reaction (PCR) detection method. Chomatography analysis of 48 isolates identified 36 as aflatoxigenic (75%). One hundred and forty-one DNA bands were generated with 11 random amplified polymorphic DNA (RAPD) primers and analysed via unweighted pair group analysis, using arithmetic means (UPGMA). Isolates grouped according to host, with differentiation of those from A. occidentale also according to geographical origin. Aspergillus flavus-specific PCR primers ASPITSF2 and ASPITSR3 were designed from ribosomal DNA internal transcribed spacers (ITS 1 and 2), and an internal amplification control was developed, to prevent false negative results. Specificity to only A. flavus was confirmed against DNA from additional aspergilli and other fungi. RAPD-based characterization differentiated isolates according to plant host. The PCR primer pair developed showed specificity to A. flavus, with a detection limit of 10 fg. Genetic variability observed in A. flavus isolates from two Brazilian agroecosystems suggested reproductive isolation. The PCR detection method developed for A. flavus represents progress towards multiplex PCR detection of aflatoxigenic and nonaflatoxigenic strains in Hazard Analysis Critical Control Point systems.

Recombinase Polymerase Amplification Compared to Real-Time Polymerase Chain Reaction Test for the Detection of Fasciola hepatica in Human Stool

PubMed Central

Cabada, Miguel M.; Malaga, Jose L.; Castellanos-Gonzalez, Alejandro; Bagwell, Kelli A.; Naeger, Patrick A.; Rogers, Hayley K.; Maharsi, Safa; Mbaka, Maryann; White, A. Clinton

2017-01-01

Fasciola hepatica is the most widely distributed trematode infection in the world. Control efforts may be hindered by the lack of diagnostic capacity especially in remote endemic areas. Polymerase chain reaction (PCR)–based methods offer high sensitivity and specificity but require expensive technology. However, the recombinase polymerase amplification (RPA) is an efficient isothermal method that eliminates the need for a thermal cycler and has a high deployment potential to resource-limited settings. We report on the characterization of RPA and PCR tests to detect Fasciola infection in clinical stool samples with low egg burdens. The sensitivity of the RPA and PCR were 87% and 66%, respectively. Both tests were 100% specific showing no cross-reactivity with trematode, cestode, or nematode parasites. In addition, RPA and PCR were able to detect 47% and 26% of infections not detected by microscopy, respectively. The RPA adapted to a lateral flow platform was more sensitive than gel-based detection of the reaction products. In conclusion, the Fasciola RPA is a highly sensitive and specific test to diagnose chronic infection using stool samples. The Fasciola RPA lateral flow has the potential for deployment to endemic areas after further characterization. PMID:27821691

SVM-based automatic diagnosis method for keratoconus

NASA Astrophysics Data System (ADS)

Gao, Yuhong; Wu, Qiang; Li, Jing; Sun, Jiande; Wan, Wenbo

2017-06-01

Keratoconus is a progressive cornea disease that can lead to serious myopia and astigmatism, or even to corneal transplantation, if it becomes worse. The early detection of keratoconus is extremely important to know and control its condition. In this paper, we propose an automatic diagnosis algorithm for keratoconus to discriminate the normal eyes and keratoconus ones. We select the parameters obtained by Oculyzer as the feature of cornea, which characterize the cornea both directly and indirectly. In our experiment, 289 normal cases and 128 keratoconus cases are divided into training and test sets respectively. Far better than other kernels, the linear kernel of SVM has sensitivity of 94.94% and specificity of 97.87% with all the parameters training in the model. In single parameter experiment of linear kernel, elevation with 92.03% sensitivity and 98.61% specificity and thickness with 97.28% sensitivity and 97.82% specificity showed their good classification abilities. Combining elevation and thickness of the cornea, the proposed method can reach 97.43% sensitivity and 99.19% specificity. The experiments demonstrate that the proposed automatic diagnosis method is feasible and reliable.

Novel α-L-arabinofuranosidase from Cellulomonas fimi ATCC 484 and its substrate-specificity analysis with the aid of computer.

PubMed

Yang, Ying; Zhang, Lujia; Guo, Mingrong; Sun, Jiaqi; Matsukawa, Shingo; Xie, Jingli; Wei, Dongzhi

2015-04-15

In the process of gene mining for novel α-L-arabinofuranosidases (AFs), the gene Celf_3321 from Cellulomonas fimi ATCC 484 encodes an AF, termed as AbfCelf, with potent activity, 19.4 U/mg under the optimum condition, pH 6.0 and 40 °C. AbfCelf can hydrolyze α-1,5-linked oligosaccharides, sugar beet arabinan, linear 1,5-α-arabinan, and wheat flour arabinoxylan, which is partly different from some previously well-characterized GH 51 AFs. The traditional substrate-specificity analysis for AFs is labor-consuming and money costing, because the substrates include over 30 kinds of various 4-nitrophenol (PNP)-glycosides, oligosaccharides, and polysaccharides. Hence, a preliminary structure and mechanism based method was applied for substrate-specificity analysis. The binding energy (ΔG, kcal/mol) obtained by docking suggested the reaction possibility and coincided with the experimental results. AbfA crystal 1QW9 was used to test the rationality of docking method in simulating the interaction between enzyme and substrate, as well the credibility of the substrate-specificity analysis method in silico.

Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy

PubMed Central

Friedrichs, Stephanie; Malan, Daniela; Voss, Yvonne; Sasse, Philipp

2015-01-01

Disease-specific induced pluripotent stem (iPS) cells can be generated from patients and differentiated into functional cardiomyocytes for characterization of the disease and for drug screening. In order to obtain pure cardiomyocytes for automated electrophysiological investigation, we here report a novel non-clonal purification strategy by using lentiviral gene transfer of a puromycin resistance gene under the control of a cardiac-specific promoter. We have applied this method to our previous reported wild-type and long QT syndrome 3 (LQTS 3)-specific mouse iPS cells and obtained a pure cardiomyocyte population. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. Action potentials and field potentials showed the characteristic prolongation at low heart rates in LQTS 3-specific, but not in wild-type iPS cell-derived cardiomyocytes. Hence, LQTS 3-specific cardiomyocytes can be purified from iPS cells with a lentiviral strategy, maintain the hallmarks of the LQTS 3 disease and can be used for automated electrophysiological characterization and drug screening. PMID:26237021

Domain-based assays of individual antibody concentrations in an oligoclonal combination targeting a single protein.

PubMed

Meng, Q; Li, M; Silberg, M A; Conrad, F; Bettencourt, J; To, R; Huang, C; Ma, J; Meyer, K; Shimizu, R; Cao, L; Tomic, M T; Marks, J D

2012-02-15

Quantitation of individual monoclonal antibodies (mAbs) within a combined antibody drug product is required for preclinical and clinical drug development, including pharmacokinetic (PK), toxicology, stability, and biochemical characterization studies of such drugs. We have developed an antitoxin, XOMA 3AB, consisting of three recombinant mAbs that potently neutralize the known subtypes of type A botulinum neurotoxin (BoNT/A). The three mAbs bind nonoverlapping BoNT/A epitopes with high affinity. XOMA 3AB is being developed as a treatment for botulism resulting from BoNT/A. To develop antibody-specific assays, we cloned, expressed, and purified BoNT/A domains from Escherichia coli. Each mAb bound only to its specific domain with affinity comparable to the binding to holotoxin. mAb-specific domains were used to develop an enzyme-linked immunosorbent assay (ELISA) for characterization of the integrity and binding activity of the three mAbs in the drug product. An electrochemiluminescence bridging assay that is robust to interference from components in serum was also developed, and we demonstrate that it can be used for PK assays. This type of antigen engineering to generate mAb-specific domains is a general method allowing quantitation and characterization of individual mAbs in a mAb cocktail that binds the same protein and is superior to anti-idiotype approaches. Copyright © 2011 Elsevier Inc. All rights reserved.

Domain based assays of individual antibody concentrations in an oligoclonal combination targeting a single protein

PubMed Central

Meng, Q.; Li, M.; Silberg, M.A.; Conrad, F.; Bettencourt, J.; To, R.; Huang, C.; Ma, J.; Meyer, K.; Shimizu, R.; Cao, L.; Tomic, M.T.; Marks, J.D.

2014-01-01

Quantitation of individual mAbs within a combined antibody drug product is required for preclinical and clinical drug development including pharmacokinetics (PK), toxicology, stability and biochemical characterization studies of such drugs. We have developed an antitoxin (XOMA 3AB) consisting of three recombinant monoclonal antibodies (mAbs) that potently neutralizes the known subtypes of type A botulinum neurotoxin (BoNT/A). The three mAbs bind non-overlapping BoNT/A epitopes with high affinity. XOMA3AB is being developed as a treatment for botulism resulting from BoNT/A. To develop antibody-specific assays, we cloned, expressed, and purified BoNT/A domains from E. coli. Each mAb bound only to its specific domain with affinity comparable to the binding to holotoxin. MAb specific domains were used to develop an ELISA for characterization of the integrity and binding activity of the three mAbs in the drug product. An electrochemiluminescence bridging assay was also developed that is robust to interference from components in serum and we demonstrate that it can be used for PK assays. This type of antigen engineering to generate mAb-specific domains is a general method allowing quantitation and characterization of individual mAbs in a mAb cocktail that bind the same protein and is superior to anti-idiotype approaches. PMID:22037290

Mentoring in Cooperative Education and Internships: Preparing Proteges for STEM Professions

ERIC Educational Resources Information Center

Fifolt, Matt; Searby, Linda

2010-01-01

This paper presents the findings of a mixed methods study regarding students' perceptions of mentoring in a cooperative education (co-op) program. Specifically, content analysis of student interviews suggested a lack of understanding of the mentoring process as characterized by unexamined assumptions and inadequate preparation for the co-op…

Aviation Security, Risk Assessment, and Risk Aversion for Public Decisionmaking

ERIC Educational Resources Information Center

Stewart, Mark G.; Mueller, John

2013-01-01

This paper estimates risk reductions for each layer of security designed to prevent commercial passenger airliners from being commandeered by terrorists, kept under control for some time, and then crashed into specific targets. Probabilistic methods are used to characterize the uncertainty of rates of deterrence, detection, and disruption, as well…

Neuropathogenic capacity of lentogenic, mesogenic, and velogenic Newcastle disease virus strains in day-old chickens

USDA-ARS?s Scientific Manuscript database

Strains of Newcastle disease virus (NDV) have different abilities to elicit neurological signs in infected birds. Although neuropathogenesis has been characterized in classical animal experiments, there is no specific “in vivo” method to study the intrinsic capacity of NDV strains to infect the bra...

Synthesis of LaVO4/TiO2 heterojunction nanotubes by sol-gel coupled with hydrothermal method for photocatalytic air purification.

PubMed

Zou, Xuejun; Li, Xinyong; Zhao, Qidong; Liu, Shaomin

2012-10-01

With the aim of improving the effective utilization of visible light, the LaVO(4)/TiO(2) heterojunction nanotubes were fabricated by sol-gel coupled with hydrothermal method. The photocatalytic ability was demonstrated through catalytic removal of gaseous toluene species. The nanotube samples were characterized by scanning electron microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), surface photovoltage (SPV), Raman spectra and N(2) adsorption-desorption measurements. The characterization results showed that the samples with high specific surface areas were of typical nanotubular morphology, which would lead to the high separation and transfer efficiency of photo induced electron-hole pairs. The as-prepared nanotubes exhibited high photocatalytic activity in decomposing toluene species under visible light irradiation with fine photochemical stability. The enhanced photocatalytic performance of LaVO(4)/TiO(2) nanotubes might be attributed to the matching band potentials, the interconnected heterojunction of LaVO(4) versus TiO(2), and the large specific surface areas of nanotubes. Copyright © 2012 Elsevier Inc. All rights reserved.

Electrophysiological characterization of recombinant and native P2X receptors.

PubMed

Niforatos, Wende; Jarvis, Michael F

2004-10-01

ATP acts as a fast neurotransmitter by activating a family of ligand-gated ion channels, the P2X receptors. Functional homomeric P2X(3) and heteromeric P2X(2/3) receptors are highly localized on primary sensory afferent neurons that transmit nociceptive sensory information. Activation of these P2X(3)-containing channels may provide a specific mechanism whereby ATP, released via synaptic transmission or by cellular injury, elicits pain. The experimental procedures described in this unit are useful for the electorphysiological characterization of P2X receptors. In addition, these protocols provide methods for the evaluation of ligands that interact with P2X receptors that are either natively expressed on excitable cells or cloned and expressed in heterologous cell systems. These methods are derived from standard electrophysiological principles and procedures that are applicable to a wide variety of ligand-gated ion channels. Specific attention is given here to the reliable electrophysiological measurement of both quickly (P2X(3)) and more slowly (P2X(2) and P2X(2/3)) desensitizing receptors.

Schizophrenia proteomics: biomarkers on the path to laboratory medicine?

PubMed Central

Lakhan, Shaheen Emmanuel

2006-01-01

Over two million Americans are afflicted with schizophrenia, a debilitating mental health disorder with a unique symptomatic and epidemiological profile. Genomics studies have hinted towards candidate schizophrenia susceptibility chromosomal loci and genes. Modern proteomic tools, particularly mass spectrometry and expression scanning, aim to identify both pathogenic-revealing and diagnostically significant biomarkers. Only a few studies on basic proteomics have been conducted for psychiatric disorders relative to the plethora of cancer specific experiments. One such proteomic utility enables the discovery of proteins and biological marker fingerprinting profiling techniques (SELDI-TOF-MS), and then subjects them to tandem mass spectrometric fragmentation and de novo protein sequencing (MALDI-TOF/TOF-MS) for the accurate identification and characterization of the proteins. Such utilities can explain the pathogenesis of neuro-psychiatric disease, provide more objective testing methods, and further demonstrate a biological basis to mental illness. Although clinical proteomics in schizophrenia have yet to reveal a biomarker with diagnostic specificity, methods that better characterize the disorder using endophenotypes can advance findings. Schizophrenia biomarkers could potentially revolutionize its psychopharmacology, changing it into a more hypothesis and genomic/proteomic-driven science. PMID:16846510

Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR.

PubMed

Ali, Shahzad; Ali, Qurban; Melzer, Falk; Khan, Iahtasham; Akhter, Shamim; Neubauer, Heinrich; Jamal, Syed M

2014-01-01

Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell's serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n = 5), aborted fetuses (n = 13), and vaginal swabs (n = 12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.

Characterization of the volatile profiles of beer using headspace solid-phase microextraction and gas chromatography-mass spectrometry.

PubMed

Rossi, Serena; Sileoni, Valeria; Perretti, Giuseppe; Marconi, Ombretta

2014-03-30

The objective of this study was a multivariate characterization of the volatile profile of beers. Such a characterization is timely considering the increasing worldwide consumption of beer, the continuous growth of microbreweries and the importance of volatile compounds to beer flavour. A method employing solid-phase microextraction and gas chromatography-mass spectrometry (SPME-GC-MS) was optimized and then applied to a sample set of 36 industrial and craft beers of various styles and fermentation types. The volatile profiles of different beer styles is described, with particular attention paid to the volatile compounds characteristic of a spontaneously fermented lambic raspberry framboise beer. Furthermore, it was also possible to identify which specific volatile compounds are principally responsible for the differences in the volatile profiles of top- and bottom-fermented beers. Moreover, a volatile fingerprint of the craft top-fermented Italian beers was defined, as they show a very similar volatile profile. Finally, the volatile compounds that are characteristic of the bock-style beers are described. The SPME-GC-MS analytical method optimized in this study is suitable for characterizing the volatile fingerprint of different beers, especially on the basis of the kind of fermentation (top, bottom or spontaneous), the method of production and the style of the beer. © 2013 Society of Chemical Industry.

Noninvasive Evaluation of Special Alloys for Prostheses Using Complementary Methods

NASA Astrophysics Data System (ADS)

Savin, A.; Vizureanu, P.; Prevorovsky, Z.; Chlada, M.; Krofta, J.; Baltatu, M. S.; Istrate, B.; Steigmann, R.

2018-06-01

Ti-Mo-Si alloys have gained the attention of biomedical industry due to specific strength and corrosion resistance and the best biocompatibility among metallic materials used in medical prostheses. In order to characterize the material, the experimental determination of elastic matrix, mechanical wear and the probability of appearance and propagation of thin cracks are imposed. Thus, resonant ultrasound spectroscopy and acoustic emission as non-invasive methods and complementary methods as SEM, EDX are involved, to choose the best concentration of elements with the aim of mechanical properties improvement.

A Retrospective Evaluation of the Use of Mass Spectrometry in FDA Biologics License Applications

NASA Astrophysics Data System (ADS)

Rogstad, Sarah; Faustino, Anneliese; Ruth, Ashley; Keire, David; Boyne, Michael; Park, Jun

2017-05-01

The characterization sections of biologics license applications (BLAs) approved by the United States Food and Drug Administration (FDA) between 2000 and 2015 were investigated to examine the extent of the use of mass spectrometry. Mass spectrometry was found to be integral to the characterization of these biotherapeutics. Of the 80 electronically submitted monoclonal antibody and protein biotherapeutic BLAs included in this study, 79 were found to use mass spectrometric workflows for protein or impurity characterization. To further examine how MS is being used in successful BLAs, the applications were filtered based on the type and number of quality attributes characterized, the mass spectrometric workflows used (peptide mapping, intact mass analysis, and cleaved glycan analysis), the methods used to introduce the proteins into the gas phase (ESI, MALDI, or LC-ESI), and the specific types of instrumentation used. Analyses were conducted over a time course based on the FDA BLA approval to determine if any trends in utilization could be observed over time. Additionally, the different classes of protein-based biotherapeutics among the approved BLAs were clustered to determine if any trends could be attributed to the specific type of biotherapeutic.

Top-down and middle-down approach by fraction collection enrichment using off-line capillary electrophoresis - mass spectrometry coupling: Application to monoclonal antibody Fc/2 charge variants.

PubMed

Biacchi, Michael; Said, Nassur; Beck, Alain; Leize-Wagner, Emmanuelle; François, Yannis-Nicolas

2017-05-19

The characterization of complex protein mixtures represents one of the biggest challenge in many research fields such as biological or biopharmaceutical sciences. Out of all categories, monoclonal antibodies (mAbs) and related products drawn the most interest due to their strong therapeutic potency and specificity. Because of their intrinsic complexity due to a large number of micro-heterogeneities, there is a crucial need for analytical methods to provide comprehensive in-depth characterization of these proteins. In this work, we developed a methodology using CE-UV/MALDI-MS to perform top-down or middle-down characterization after fraction collection enrichment applied to intact protein and mAbs samples. The performance of the method was evaluated with the rapid separation of three intact protein mixture. Good robustness of CZE separation and quality of MALDI-MS spectra and MALDI-ISD spectra of each protein confirms the usefulness of sample enrichment to obtain adequate quantity of deposed protein for top-down analysis and the proof of principle of the method. In a second step, the method was applied to the middle-down characterization of Fc/2 cetuximab variants. Identification of around 9% sequence coverage of Fc/2 cetuximab fragments allows to conclude on the feasibility of the strategy for middle-down characterization of Fc/2 cetuximab variants using CE-UV/MALDI-MS. Moreover, MALDI-ISD fragmentation of Fc/2 cetuximab variants confirm separation phenomenon based on the formation of Fc/2 dimers with and without C-terminal truncation. Copyright © 2017 Elsevier B.V. All rights reserved.

Multipole Vortex Blobs (MVB): Symplectic Geometry and Dynamics.

PubMed

Holm, Darryl D; Jacobs, Henry O

2017-01-01

Vortex blob methods are typically characterized by a regularization length scale, below which the dynamics are trivial for isolated blobs. In this article, we observe that the dynamics need not be trivial if one is willing to consider distributional derivatives of Dirac delta functionals as valid vorticity distributions. More specifically, a new singular vortex theory is presented for regularized Euler fluid equations of ideal incompressible flow in the plane. We determine the conditions under which such regularized Euler fluid equations may admit vorticity singularities which are stronger than delta functions, e.g., derivatives of delta functions. We also describe the symplectic geometry associated with these augmented vortex structures, and we characterize the dynamics as Hamiltonian. Applications to the design of numerical methods similar to vortex blob methods are also discussed. Such findings illuminate the rich dynamics which occur below the regularization length scale and enlighten our perspective on the potential for regularized fluid models to capture multiscale phenomena.

Discovery, detection and use of biomarkers

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

Swanson, Basil I.; Mukundan, Harshini; Sakamuri, Rama Murthy

Provided herein are systems for and methods of capturing, detecting, quantifying, and characterizing target moieties that are characterized by having a lipophilic portion of sufficient size and chemical composition whereby the target moiety inserts (or partitions) into a lipid assembly. Examples of such assays employ synthetic lipid constructs such as supported bilayers which are used to capture target moieties; other example assays exploit the natural absorption of compounds into natural lipid constructs such as HDL or LDL particles or cell membranes to capture target moieties. In specific embodiments, the target moieties are bacterial pathogen associated molecular pattern (PAMP) molecules ormore » compounds not yet identified as PAMP molecules. Also provided are methods of determining PAMP molecule fingerprints and profiles that are linked to (indicative of) bacterial infection, disease states or progression, development of antibiotic resistance, and so forth, as well as these fingerprints, profiles and methods of using them.« less

Hierarchical Ensemble Methods for Protein Function Prediction

PubMed Central

2014-01-01

Protein function prediction is a complex multiclass multilabel classification problem, characterized by multiple issues such as the incompleteness of the available annotations, the integration of multiple sources of high dimensional biomolecular data, the unbalance of several functional classes, and the difficulty of univocally determining negative examples. Moreover, the hierarchical relationships between functional classes that characterize both the Gene Ontology and FunCat taxonomies motivate the development of hierarchy-aware prediction methods that showed significantly better performances than hierarchical-unaware “flat” prediction methods. In this paper, we provide a comprehensive review of hierarchical methods for protein function prediction based on ensembles of learning machines. According to this general approach, a separate learning machine is trained to learn a specific functional term and then the resulting predictions are assembled in a “consensus” ensemble decision, taking into account the hierarchical relationships between classes. The main hierarchical ensemble methods proposed in the literature are discussed in the context of existing computational methods for protein function prediction, highlighting their characteristics, advantages, and limitations. Open problems of this exciting research area of computational biology are finally considered, outlining novel perspectives for future research. PMID:25937954

Quantitative characterization of conformational-specific protein-DNA binding using a dual-spectral interferometric imaging biosensor.

PubMed

Zhang, Xirui; Daaboul, George G; Spuhler, Philipp S; Dröge, Peter; Ünlü, M Selim

2016-03-14

DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are not fully understood. Recently, it was discovered that DNA-binding proteins recognize specific binding sites to carry out their functions through an indirect readout mechanism by recognizing and capturing DNA conformational flexibility and deformation. High-throughput DNA microarray-based methods that provide large-scale protein-DNA binding information have shown effective and comprehensive analysis of protein-DNA binding affinities, but do not provide information of DNA conformational changes in specific protein-DNA complexes. Building on the high-throughput capability of DNA microarrays, we demonstrate a quantitative approach that simultaneously measures the amount of protein binding to DNA and nanometer-scale DNA conformational change induced by protein binding in a microarray format. Both measurements rely on spectral interferometry on a layered substrate using a single optical instrument in two distinct modalities. In the first modality, we quantitate the amount of binding of protein to surface-immobilized DNA in each DNA spot using a label-free spectral reflectivity technique that accurately measures the surface densities of protein and DNA accumulated on the substrate. In the second modality, for each DNA spot, we simultaneously measure DNA conformational change using a fluorescence vertical sectioning technique that determines average axial height of fluorophores tagged to specific nucleotides of the surface-immobilized DNA. The approach presented in this paper, when combined with current high-throughput DNA microarray-based technologies, has the potential to serve as a rapid and simple method for quantitative and large-scale characterization of conformational specific protein-DNA interactions.

Modelling based on Spatial Impulse Response Model for Optimization of Inter Digital Transducers (SAW Sensors) for Non Destructive Testing

NASA Astrophysics Data System (ADS)

Fall, D.; Duquennoy, M.; Ouaftouh, M.; Piwakowski, B.; Jenot, F.

This study deals with modelling SAW-IDT transducers for their optimization. These sensors are specifically developed to characterize properties of thin layers, coatings and functional surfaces. Among the methods of characterization, the ultrasonic methods using Rayleigh surface waves are particularly interesting because the propagation of these waves is close to the surface of material and the energy is concentrated within a layer under the surface of about one wavelength thick. In order to characterize these coatings and structures, it is necessary to work in high frequencies, this is why in this study, SAW-IDT sensors are realized for surface acoustic wave generation. For optimization of these SAW-IDT sensors, particularly their band-width, it is necessary to study various IDT configurations by varying the number of electrodes, dimensions of the electrodes, their shapes and spacings. Thus it is necessary to implement effective and rapid technique for modelling. The originality of this study is to develop simulation tools based on Spatial Impulse Response model. Therefore it will be possible to reduce considerably computing time and results are obtained in a few seconds, instead of several hours (or days) by using finite element method. In order to validate this method, theoretical and experimental results are compared with finite element method and Interferometric measurements. The results obtained show a good overall concordance and confirm effectiveness of suggested method.

Generation of Regionally Specific Neural Progenitor Cells (NPCs) and Neurons from Human Pluripotent Stem Cells (hPSCs).

PubMed

Cutts, Josh; Brookhouser, Nicholas; Brafman, David A

2016-01-01

Neural progenitor cells (NPCs) derived from human pluripotent stem cells (hPSCs) are a multipotent cell population capable of long-term expansion and differentiation into a variety of neuronal subtypes. As such, NPCs have tremendous potential for disease modeling, drug screening, and regenerative medicine. Current methods for the generation of NPCs results in cell populations homogenous for pan-neural markers such as SOX1 and SOX2 but heterogeneous with respect to regional identity. In order to use NPCs and their neuronal derivatives to investigate mechanisms of neurological disorders and develop more physiologically relevant disease models, methods for generation of regionally specific NPCs and neurons are needed. Here, we describe a protocol in which exogenous manipulation of WNT signaling, through either activation or inhibition, during neural differentiation of hPSCs, promotes the formation of regionally homogenous NPCs and neuronal cultures. In addition, we provide methods to monitor and characterize the efficiency of hPSC differentiation to these regionally specific cell identities.

Allele-specific copy-number discovery from whole-genome and whole-exome sequencing

PubMed Central

Wang, WeiBo; Wang, Wei; Sun, Wei; Crowley, James J.; Szatkiewicz, Jin P.

2015-01-01

Copy-number variants (CNVs) are a major form of genetic variation and a risk factor for various human diseases, so it is crucial to accurately detect and characterize them. It is conceivable that allele-specific reads from high-throughput sequencing data could be leveraged to both enhance CNV detection and produce allele-specific copy number (ASCN) calls. Although statistical methods have been developed to detect CNVs using whole-genome sequence (WGS) and/or whole-exome sequence (WES) data, information from allele-specific read counts has not yet been adequately exploited. In this paper, we develop an integrated method, called AS-GENSENG, which incorporates allele-specific read counts in CNV detection and estimates ASCN using either WGS or WES data. To evaluate the performance of AS-GENSENG, we conducted extensive simulations, generated empirical data using existing WGS and WES data sets and validated predicted CNVs using an independent methodology. We conclude that AS-GENSENG not only predicts accurate ASCN calls but also improves the accuracy of total copy number calls, owing to its unique ability to exploit information from both total and allele-specific read counts while accounting for various experimental biases in sequence data. Our novel, user-friendly and computationally efficient method and a complete analytic protocol is freely available at https://sourceforge.net/projects/asgenseng/. PMID:25883151

Applications of Fluorescence Spectroscopy for dissolved organic matter characterization in wastewater treatment plants

NASA Astrophysics Data System (ADS)

Goffin, Angélique; Guérin, Sabrina; Rocher, Vincent; Varrault, Gilles

2016-04-01

Dissolved organic matter (DOM) influences wastewater treatment plants efficiency (WTTP): variations in its quality and quantity can induce a foaming phenomenon and a fouling event inside biofiltration processes. Moreover, in order to manage denitrification step (control and optimization of the nitrate recirculation), it is important to be able to estimate biodegradable organic matter quantity before biological treatment. But the current methods used to characterize organic matter quality, like biological oxygen demand are laborious, time consuming and sometimes not applicable to directly monitor organic matter in situ. In the context of MOCOPEE research program (www.mocopee.com), this study aims to assess the use of optical techniques, such as UV-Visible absorbance and more specifically fluorescence spectroscopy in order to monitor and to optimize process efficiency in WWTP. Fluorescence excitation-emission matrix (EEM) spectroscopy was employed to prospect the possibility of using this technology online and in real time to characterize dissolved organic matter in different effluents of the WWTP Seine Centre (240,000 m3/day) in Paris, France. 35 sewage water influent samples were collected on 10 days at different hours. Data treatment were performed by two methods: peak picking and parallel factor analysis (PARAFAC). An evolution of DOM quality (position of excitation - emission peaks) and quantity (intensity of fluorescence) was observed between the different treatment steps (influent, primary treatment, biological treatment, effluent). Correlations were found between fluorescence indicators and different water quality key parameters in the sewage influents. We developed different multivariate linear regression models in order to predict a variety of water quality parameters by fluorescence intensity at specific excitation-emission wavelengths. For example dissolved biological oxygen demand (r2=0,900; p<0,0001) and ammonium concentration (r2=0,898; p<0,0001) present good correlation with specific fluorescence peaks and indicators. These indicators derived from 3D spectrofluorescence could be used in order to characterize DOM online and thus to optimize process efficiency in WWTP.

A system for online beam emittance measurements and proton beam characterization

NASA Astrophysics Data System (ADS)

Nesteruk, K. P.; Auger, M.; Braccini, S.; Carzaniga, T. S.; Ereditato, A.; Scampoli, P.

2018-01-01

A system for online measurement of the transverse beam emittance was developed. It is named 4PrOBεaM (4-Profiler Online Beam Emittance Measurement) and was conceived to measure the emittance in a fast and efficient way using the multiple beam profiler method. The core of the system is constituted by four consecutive UniBEaM profilers, which are based on silica fibers passing across the beam. The 4PrOBεaM system was deployed for characterization studies of the 18 MeV proton beam produced by the IBA Cyclone 18 MeV cyclotron at Bern University Hospital (Inselspital). The machine serves daily radioisotope production and multi-disciplinary research, which is carried out with a specifically conceived Beam Transport Line (BTL). The transverse RMS beam emittance of the cyclotron was measured as a function of several machine parameters, such as the magnetic field, RF peak voltage, and azimuthal angle of the stripper. The beam emittance was also measured using the method based on the quadrupole strength variation. The results obtained with both techniques were compared and a good agreement was found. In order to characterize the longitudinal dynamics, the proton energy distribution was measured. For this purpose, a method was developed based on aluminum absorbers of different thicknesses, a UniBEaM detector, and a Faraday cup. The results were an input for a simulation of the BTL developed in the MAD-X software. This tool allows machine parameters to be tuned online and the beam characteristics to be optimized for specific applications.

Characterizability of metabolic pathway systems from time series data.

PubMed

Voit, Eberhard O

2013-12-01

Over the past decade, the biomathematical community has devoted substantial effort to the complicated challenge of estimating parameter values for biological systems models. An even more difficult issue is the characterization of functional forms for the processes that govern these systems. Most parameter estimation approaches tacitly assume that these forms are known or can be assumed with some validity. However, this assumption is not always true. The recently proposed method of Dynamic Flux Estimation (DFE) addresses this problem in a genuinely novel fashion for metabolic pathway systems. Specifically, DFE allows the characterization of fluxes within such systems through an analysis of metabolic time series data. Its main drawback is the fact that DFE can only directly be applied if the pathway system contains as many metabolites as unknown fluxes. This situation is unfortunately rare. To overcome this roadblock, earlier work in this field had proposed strategies for augmenting the set of unknown fluxes with independent kinetic information, which however is not always available. Employing Moore-Penrose pseudo-inverse methods of linear algebra, the present article discusses an approach for characterizing fluxes from metabolic time series data that is applicable even if the pathway system is underdetermined and contains more fluxes than metabolites. Intriguingly, this approach is independent of a specific modeling framework and unaffected by noise in the experimental time series data. The results reveal whether any fluxes may be characterized and, if so, which subset is characterizable. They also help with the identification of fluxes that, if they could be determined independently, would allow the application of DFE. Copyright © 2013 Elsevier Inc. All rights reserved.

Characterizability of Metabolic Pathway Systems from Time Series Data

PubMed Central

Voit, Eberhard O.

2013-01-01

Over the past decade, the biomathematical community has devoted substantial effort to the complicated challenge of estimating parameter values for biological systems models. An even more difficult issue is the characterization of functional forms for the processes that govern these systems. Most parameter estimation approaches tacitly assume that these forms are known or can be assumed with some validity. However, this assumption is not always true. The recently proposed method of Dynamic Flux Estimation (DFE) addresses this problem in a genuinely novel fashion for metabolic pathway systems. Specifically, DFE allows the characterization of fluxes within such systems through an analysis of metabolic time series data. Its main drawback is the fact that DFE can only directly be applied if the pathway system contains as many metabolites as unknown fluxes. This situation is unfortunately rare. To overcome this roadblock, earlier work in this field had proposed strategies for augmenting the set of unknown fluxes with independent kinetic information, which however is not always available. Employing Moore-Penrose pseudo-inverse methods of linear algebra, the present article discusses an approach for characterizing fluxes from metabolic time series data that is applicable even if the pathway system is underdetermined and contains more fluxes than metabolites. Intriguingly, this approach is independent of a specific modeling framework and unaffected by noise in the experimental time series data. The results reveal whether any fluxes may be characterized and, if so, which subset is characterizable. They also help with the identification of fluxes that, if they could be determined independently, would allow the application of DFE. PMID:23391489

Characterization of Olive Oil by Ultrasonic and Physico-chemical Methods

NASA Astrophysics Data System (ADS)

Alouache, B.; Khechena, F. K.; Lecheb, F.; Boutkedjirt, T.

Olive oil excels by its nutritional and medicinal benefits. It can be consumed without any treatment. However, its quality can be altered by inadequate storage conditions or if it is mixed with other kinds of oils. The objective of this work is to demonstrate the ability of ultrasonic methods to characterize and control olive oil quality. By using of a transducer of 2.25 MHz nominal frequency, in pulse echo mode, ultrasonic parameters, such as propagation velocity and attenuation,have been measured for pure olive oil and for its mixtures with sunflower oil at different proportions. Mechanical properties, such as density and viscosity, have also been determined. The results of ultrasonic measurements are consistent with those obtained by physico-chemical methods, such as rancidity degree, acid index, UV specific extinction coefficient and viscosity. They show that the ultrasonic method allows to distinguish between mixtures at different proportions. The study allows concluding that ultrasound techniques can be considered as a useful complement to existing physico-chemical analysis techniques.

Fabrication and characterization of magnesium and calcium trimesate complexes via ion-exchange and one-pot self-assembly reaction

NASA Astrophysics Data System (ADS)

Ozer, Demet; Oztas, Nursen Altuntas; Köse, Dursun A.; Şahin, Onur

2018-03-01

Using two different synthesis methods, two diversified magnesium and calcium complexes were successfully prepared. When the ion exchange method was used, C9H14MgO11.H2O and C18H30Ca3O24 complexes were obtained. When the one-pot self-assembly reaction was used, C18H34Mg3O26.4H2O and C9H12CaO10 complexes were produced. The structural characterizations were performed by using X-ray diffraction, FT-IR and elemental analyses. Thermal behavior of complexes were also determined via TGA method. The both complexes of magnesium and calcium trimesate have micro and mesoporosity with low porosity because of hydrogen bonds. Then hydrogen storage capacities of complexes were also determined. The differences in synthesis method result in the differences on complexes structure, morphology (shape, particle size and specific surface area) and hydrogen storage capacities.

Surface Dilution Kinetics Using Substrate Analog-Enantiomers as Diluents: Enzymatic Lipolysis by Bee-Venom Phospholipase A2

PubMed Central

Singh, Jasmeet; Ranganathan, Radha; Hajdu, Joseph

2010-01-01

A novel assay employing D-enantiomers of phospholipids as diluents for characterizing surface kinetics of lipid hydrolysis by phospholipases is introduced. The rationale of the method are: (i) D-enantiomers resist hydrolysis because of the stereoselectivity of the enzymes toward L-enantiomers and (ii) mixtures of L+D-lipids at various L:D ratios but constant L+D-lipid concentrations yield a surface dilution series of variable L-lipid concentration with constant medium properties. Kinetic characterization of bee-venom phospholipase A2 activity at bile salt + phospholipid aggregate-water interfaces was performed using the mixed L+D-lipid surface dilution assay and interface kinetic parameters were obtained. The assay applies to bio-membrane models as well. Activity was measured by pH-Stat methods. Aggregation numbers and interface hydration/microviscosity measured by time resolved fluorescence quenching and electron spin resonance respectively confirmed that interface properties were indeed invariant in a surface dilution series, supporting rationale (ii) and were used to calculate substrate concentrations. Activity data show excellent agreement with a kinetic model derived with D-enantiomers as diluents and also that D-phospholipids bind to the enzyme but resist hydrolysis; underscoring rationale (i). The assay is significant to enabling determination of interface specific kinetic parameters for the first time and thereby characterization of interface specificity of lipolytic enzymes. PMID:20727845

Invasive candidiasis: future directions in non-culture based diagnosis.

PubMed

Posch, Wilfried; Heimdörfer, David; Wilflingseder, Doris; Lass-Flörl, Cornelia

2017-09-01

Delayed initial antifungal therapy is associated with high mortality rates caused by invasive candida infections, since accurate detection of the opportunistic pathogenic yeast and its identification display a diagnostic challenge. diagnosis of candida infections relies on time-consuming methods such as blood cultures, serologic and histopathologic examination. to allow for fast detection and characterization of invasive candidiasis, there is a need to improve diagnostic tools. trends in diagnostics switch to non-culture-based methods, which allow specified diagnosis within significantly shorter periods of time in order to provide early and appropriate antifungal treatment. Areas covered: within this review comprise novel pathogen- and host-related testing methods, e.g. multiplex-PCR analyses, T2 magnetic resonance, fungus-specific DNA microarrays, microRNA characterization or analyses of IL-17 as biomarker for early detection of invasive candidiasis. Expert commentary: Early recognition and diagnosis of fungal infections is a key issue for improved patient management. As shown in this review, a broad range of novel molecular based tests for the detection and identification of Candida species is available. However, several assays are in-house assays and lack standardization, clinical validation as well as data on sensitivity and specificity. This underscores the need for the development of faster and more accurate diagnostic tests.

Systematic Biological Filter Design with a Desired I/O Filtering Response Based on Promoter-RBS Libraries.

PubMed

Hsu, Chih-Yuan; Pan, Zhen-Ming; Hu, Rei-Hsing; Chang, Chih-Chun; Cheng, Hsiao-Chun; Lin, Che; Chen, Bor-Sen

2015-01-01

In this study, robust biological filters with an external control to match a desired input/output (I/O) filtering response are engineered based on the well-characterized promoter-RBS libraries and a cascade gene circuit topology. In the field of synthetic biology, the biological filter system serves as a powerful detector or sensor to sense different molecular signals and produces a specific output response only if the concentration of the input molecular signal is higher or lower than a specified threshold. The proposed systematic design method of robust biological filters is summarized into three steps. Firstly, several well-characterized promoter-RBS libraries are established for biological filter design by identifying and collecting the quantitative and qualitative characteristics of their promoter-RBS components via nonlinear parameter estimation method. Then, the topology of synthetic biological filter is decomposed into three cascade gene regulatory modules, and an appropriate promoter-RBS library is selected for each module to achieve the desired I/O specification of a biological filter. Finally, based on the proposed systematic method, a robust externally tunable biological filter is engineered by searching the promoter-RBS component libraries and a control inducer concentration library to achieve the optimal reference match for the specified I/O filtering response.

Specific RNA-protein interactions detected with saturation transfer difference NMR.

PubMed

Harris, Kimberly A; Shekhtman, Alexander; Agris, Paul F

2013-08-01

RNA, at the forefront of biochemical research due to its central role in biology, is recognized by proteins through various mechanisms. Analysis of the RNA-protein interface provides insight into the recognition determinants and function. As such, there is a demand for developing new methods to characterize RNA-protein interactions. Saturation transfer difference (STD) NMR can identify binding ligands for proteins in a rather short period of time, with data acquisitions of just a few hours. Two RNA-protein systems involved in RNA modification were studied using STD NMR. The N (6)-threonylcarbamoyltransferase, YrdC, with nucleoside-specific recognition, was shown to bind the anticodon stem-loop of tRNA(Lys)UUU. The points of contact on the RNA were assigned and a binding interface was identified. STD NMR was also applied to the interaction of the archaeal ribosomal protein, L7Ae, with the box C/D K-turn RNA. The distinctiveness of the two RNA-protein interfaces was evident. Both RNAs exhibited strong STD signals indicative of direct contact with the respective protein, but reflected the nature of recognition. Characterization of nucleic acid recognition determinants traditionally involves cost and time prohibitive methods. This approach offers significant insight into interaction interfaces fairly rapidly, and complements existing structural methods.

Results and lessons learned from MODIS polarization sensitivity characterization

NASA Astrophysics Data System (ADS)

Sun, J.; Xiong, X.; Wang, X.; Qiu, S.; Xiong, S.; Waluschka, E.

2006-08-01

In addition to radiometric, spatial, and spectral calibration requirements, MODIS design specifications include polarization sensitivity requirements of less than 2% for all Reflective Solar Bands (RSB) except for the band centered at 412nm. To the best of our knowledge, MODIS was the first imaging radiometer that went through comprehensive system level (end-to-end) polarization characterization. MODIS polarization sensitivity was measured pre-launch at a number of sensor view angles using a laboratory Polarization Source Assembly (PSA) that consists of a rotatable source, a polarizer (Ahrens prism design), and a collimator. This paper describes MODIS polarization characterization approaches used by MODIS Characterization Support Team (MCST) at NASA/GSFC and addresses issues and concerns in the measurements. Results (polarization factor and phase angle) using different analyzing methods are discussed. Also included in this paper is a polarization characterization comparison between Terra and Aqua MODIS. Our previous and recent analysis of MODIS RSB polarization sensitivity could provide useful information for future Earth-observing sensor design, development, and characterization.

Separation and characterization of silybin, isosilybin, silydianin and silychristin in milk thistle extract by liquid chromatography-electrospray tandem mass spectrometry.

PubMed

Lee, James I; Hsu, Bih H; Wu, Di; Barrett, Jeffrey S

2006-05-26

A selective and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed for the characterization of silymarin in commercially available milk thistle extract. In this study, six main active constituents, including silydianin, silychristin, diastereomers of silybin (silybin A and B) and diastereomers of isosilybin (isosilybin A and B) in silymarin, were completely separated on a YMC ODS-AQ HPLC column using a gradient mobile phase system comprised of ammonium acetate and methanol/water/formic acid. Identification and characterization of the major constituents were based not only on the product ion scan, which provided unique fragmentation information of a selected molecular ion, but also on the specific fragmentation of multiple reaction monitoring (MRM) data, which confirmed the retention times of LC chromatographic peaks. The method was applied in the analysis of human plasma samples in the presence of silymarin and appeared to be suitable for the pharmacokinetic studies in which the discrimination of silymarin constituents is essential.

Scattering of circumferential waves in a cracked annulus

NASA Astrophysics Data System (ADS)

Valle, Christine; Qu, Jianmin; Jacobs, Laurence J.

2000-05-01

This paper considers guided waves propagating in the circumferential direction of an annulus with a radial crack, with the objective of developing an ultrasonic technique that can detect and characterize these cracks. Specifically, the finite element method is used to simulate the propagation and scattering of guided circumferential waves in a cracked annulus. This method fosters a better understanding of the wave fields, so that a transducer configuration used in the field can be optimized for crack detection/characterization. Both a point source (simulating laser generated ultrasound) and a distributed source (simulating a PZT transducer) are modeled and compared to corresponding experimental results. Animations (snapshots at different instants in time) of the strain energy field in the annulus are given for various combinations of load profiles, incident angles, and incident frequencies. Results of this paper provide the necessary design guidelines for developing nondestructive ultrasonic techniques for the detection/characterization of radial cracks in cylindrical pressure vessels, gas/oil pipes, and shaft/bearing systems.

Integrated Method for Purification and Single-Particle Characterization of Lentiviral Vector Systems by Size Exclusion Chromatography and Tunable Resistive Pulse Sensing.

PubMed

Heider, Susanne; Muzard, Julien; Zaruba, Marianne; Metzner, Christoph

2017-07-01

Elements derived from lentiviral particles such as viral vectors or virus-like particles are commonly used for biotechnological and biomedical applications, for example in mammalian protein expression, gene delivery or therapy, and vaccine development. Preparations of high purity are necessary in most cases, especially for clinical applications. For purification, a wide range of methods are available, from density gradient centrifugation to affinity chromatography. In this study we have employed size exclusion columns specifically designed for the easy purification of extracellular vesicles including exosomes. In addition to viral marker protein and total protein analysis, a well-established single-particle characterization technology, termed tunable resistive pulse sensing, was employed to analyze fractions of highest particle load and purity and characterize the preparations by size and surface charge/electrophoretic mobility. With this study, we propose an integrated platform combining size exclusion chromatography and tunable resistive pulse sensing for monitoring production and purification of viral particles.

Formulation and Solid State Characterization of Nicotinamide-based Co-crystals of Fenofibrate

PubMed Central

Shewale, Sheetal; Shete, A. S.; Doijad, R. C.; Kadam, S. S.; Patil, V. A.; Yadav, A. V.

2015-01-01

The present investigation deals with formulation of nicotinamide-based co-crystals of fenofibrate by different methods and solid-state characterization of the prepared co-crystals. Fenofibrate and nicotinamide as a coformer in 1:1 molar ratio were used to formulate molecular complexes by kneading, solution crystallization, antisolvent addition and solvent drop grinding methods. The prepared molecular complexes were characterized by powder X-ray diffractometry, differential scanning calorimetry, Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy and in vitro dissolution study. Considerable improvement in the dissolution rate of fenofibrate from optimized co-crystal formulation was due to an increased solubility that is attributed to the super saturation from the fine co-crystals is faster because of large specific surface area of small particles and prevention of phase transformation to pure fenofibrate. In vitro dissolution study showed that the formation of co-crystals improves the dissolution rate of fenofibrate. Nicotinamide forms the co-crystals with fenofibrate, theoretically and practically. PMID:26180279

Template-DTW based on inertial signals: Preliminary results for step characterization.

PubMed

Mantilla, Juan; Oudre, Laurent; Barrois, Remi; Vienne, Alienor; Ricard, Damien

2017-07-01

In this paper, we present a method for the creation of a library of inertial signals based on Dynamic Time Warping (DTW) for step characterization, with preliminary results in control subjects and patients with neurological diseases. Subjects performed a protocol including a 10 m straight walking, then turn back and walking for additional 10 m. The library is constructed with inertial signals (acceleration and angular velocities recorded in three directions) aligned with the DTW. Templates in the library are obtained for a specific cohort and for the different walking phases of the protocol. They are compared to the signal of a single subject by calculating a Pearson correlation coefficient. The method has been tested on a database of 864 exercises, obtained from 71 healthy controls, 24 patients with Parkinson disease and 48 patients with Radiation Induced Leukoencephalopathy (RIL). Pearson correlation classification reports a precision of about 85% for step detection. For exercise characterization the sensitivity is about 57%, 56% and 82% for Parkinson, RIL and control subjects respectively.

Enhanced Characterization of Niobium Surface Topography

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

Chen Xu, Hui Tian, Charles Reece, Michael Kelley

2011-12-01

Surface topography characterization is a continuing issue for the Superconducting Radio Frequency (SRF) particle accelerator community. Efforts are underway to both to improve surface topography, and its characterization and analysis using various techniques. In measurement of topography, Power Spectral Density (PSD) is a promising method to quantify typical surface parameters and develop scale-specific interpretations. PSD can also be used to indicate how chemical processes modifiesy the roughnesstopography at different scales. However, generating an accurate and meaningful topographic PSD of an SRF surface requires careful analysis and optimization. In this report, polycrystalline surfaces with different process histories are sampled with AFMmore » and stylus/white light interferometer profilometryers and analyzed to indicate trace topography evolution at different scales. evolving during etching or polishing. Moreover, Aan optimized PSD analysis protocol will be offered to serve the SRF surface characterization needs is presented.« less

Detection and characterization of nonspecific, sparsely-populated binding modes in the early stages of complexation

PubMed Central

Cardone, A.; Bornstein, A.; Pant, H. C.; Brady, M.; Sriram, R.; Hassan, S. A.

2015-01-01

A method is proposed to study protein-ligand binding in a system governed by specific and non-specific interactions. Strong associations lead to narrow distributions in the proteins configuration space; weak and ultra-weak associations lead instead to broader distributions, a manifestation of non-specific, sparsely-populated binding modes with multiple interfaces. The method is based on the notion that a discrete set of preferential first-encounter modes are metastable states from which stable (pre-relaxation) complexes at equilibrium evolve. The method can be used to explore alternative pathways of complexation with statistical significance and can be integrated into a general algorithm to study protein interaction networks. The method is applied to a peptide-protein complex. The peptide adopts several low-population conformers and binds in a variety of modes with a broad range of affinities. The system is thus well suited to analyze general features of binding, including conformational selection, multiplicity of binding modes, and nonspecific interactions, and to illustrate how the method can be applied to study these problems systematically. The equilibrium distributions can be used to generate biasing functions for simulations of multiprotein systems from which bulk thermodynamic quantities can be calculated. PMID:25782918

Chemometrics Methods for Specificity, Authenticity and Traceability Analysis of Olive Oils: Principles, Classifications and Applications.

PubMed

Messai, Habib; Farman, Muhammad; Sarraj-Laabidi, Abir; Hammami-Semmar, Asma; Semmar, Nabil

2016-11-17

Olive oils (OOs) show high chemical variability due to several factors of genetic, environmental and anthropic types. Genetic and environmental factors are responsible for natural compositions and polymorphic diversification resulting in different varietal patterns and phenotypes. Anthropic factors, however, are at the origin of different blends' preparation leading to normative, labelled or adulterated commercial products. Control of complex OO samples requires their (i) characterization by specific markers; (ii) authentication by fingerprint patterns; and (iii) monitoring by traceability analysis. These quality control and management aims require the use of several multivariate statistical tools: specificity highlighting requires ordination methods; authentication checking calls for classification and pattern recognition methods; traceability analysis implies the use of network-based approaches able to separate or extract mixed information and memorized signals from complex matrices. This chapter presents a review of different chemometrics methods applied for the control of OO variability from metabolic and physical-chemical measured characteristics. The different chemometrics methods are illustrated by different study cases on monovarietal and blended OO originated from different countries. Chemometrics tools offer multiple ways for quantitative evaluations and qualitative control of complex chemical variability of OO in relation to several intrinsic and extrinsic factors.

Characterization of low concentration uranium glass working materials

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

Eppich, G. R.; Wimpenny, J. B.; Leever, M. E.

A series of uranium-doped silicate glasses were created at (Lawrence Livermore National Laboratory) LLNL, to be used as working reference material analogs for low uranium concentration research. Specifically, the aim of this effort was the generation of well-characterized glasses spanning a range of concentrations and compositions, and of sufficient homogeneity in uranium concentration and isotopic composition, for instrumentation research and development purposes. While the glasses produced here are not intended to replace or become standard materials for uranium concentration or uranium isotopic composition, it is hoped that they will help fill a current gap, providing low-level uranium glasses sufficient formore » methods development and method comparisons within the limitations of the produced glass suite. Glasses are available for research use by request.« less

Annual distribution of allergenic fungal spores in atmospheric particulate matter in the Eastern Mediterranean; a comparative study between ergosterol and quantitative PCR analysis

NASA Astrophysics Data System (ADS)

Lang-Yona, N.; Dannemiller, K.; Yamamoto, N.; Burshtein, N.; Peccia, J.; Yarden, O.; Rudich, Y.

2012-03-01

Airborne fungal spores are an important fraction of atmospheric particulate matter and are major causative agents of allergenic and infectious diseases. Predicting the variability and species of allergy-causing fungal spores requires detailed and reliable methods for identification and quantification. There are diverse methods for their detection in the atmosphere and in the indoor environments; yet, it is important to optimize suitable methods for characterization of fungal spores in atmospheric samples. In this study we sampled and characterized total and specific airborne fungal spores from PM10 samples collected in Rehovot, Israel over an entire year. The total fungal spore concentrations vary throughout the year although the species variability was nearly the same. Seasonal equivalent spore concentrations analyzed by real-time quantitative-PCR-based methods were fall > winter > spring > summer. Reported concentrations based on ergosterol analysis for the same samples were and fall > spring > winter > summer. Correlation between the two analytical methods was found only for the spring season. These poor associations may be due to the per-spore ergosterol variations that arise from both varying production rates, as well as molecular degradation of ergosterol. While conversion of genome copies to spore concentration is not yet straightforward, the potential for improving this conversion and the ability of qPCR to identify groups of fungi or specific species makes this method preferable for environmental spore quantification. Identifying tools for establishing the relation between the presence of species and the actual ability to induce allergies is still needed in order to predict the effect on human health.

Annual distribution of allergenic fungal spores in atmospheric particulate matter in the eastern mediterranean; a comparative study between ergosterol and quantitative PCR analysis

NASA Astrophysics Data System (ADS)

Lang-Yona, N.; Dannemiller, K.; Yamamoto, N.; Burshtein, N.; Peccia, J.; Yarden, O.; Rudich, Y.

2011-10-01

Airborne fungal spores are an important fraction of atmospheric particulate matter and are major causative agents of allergenic and infectious diseases. Predicting the variability and species of allergy-causing fungal spores requires detailed and reliable methods for identification and quantification. There are diverse methods for their detection in the atmosphere and in the indoor environments; yet, it is important to optimize suitable methods for characterization of fungal spores in atmospheric samples. In this study we sampled and characterized total and specific airborne fungal spores from PM10 samples collected in Rohovot, Israel over an entire year. The total fungal spore concentrations vary throughout the year although the species variability was nearly the same. Seasonal equivalent spore concentrations analyzed by real-time quantitative-PCR-based methods were fall > winter > spring > summer. Reported concentrations based on ergosterol analysis for the same samples were and fall > spring > winter > summer. Correlation between the two analytical methods was found only for the spring season. These poor associations may be due to the per-spore ergosterol variations that arise from both varying production rates, as well as molecular degradation of ergosterol. While conversion of genome copies to spore concentration is not yet straightforward, the potential for improving this conversion and the ability of qPCR to identify groups of fungi or specific species makes this method preferable for environmental spore quantification. Identifying tools for establishing the relation between the presence of species and the actual ability to induce allergies is still needed in order to predict the effect on human health.

Characterization of Adrenal Adenoma by Gaussian Model-Based Algorithm.

PubMed

Hsu, Larson D; Wang, Carolyn L; Clark, Toshimasa J

2016-01-01

We confirmed that computed tomography (CT) attenuation values of pixels in an adrenal nodule approximate a Gaussian distribution. Building on this and the previously described histogram analysis method, we created an algorithm that uses mean and standard deviation to estimate the percentage of negative attenuation pixels in an adrenal nodule, thereby allowing differentiation of adenomas and nonadenomas. The institutional review board approved both components of this study in which we developed and then validated our criteria. In the first, we retrospectively assessed CT attenuation values of adrenal nodules for normality using a 2-sample Kolmogorov-Smirnov test. In the second, we evaluated a separate cohort of patients with adrenal nodules using both the conventional 10HU unit mean attenuation method and our Gaussian model-based algorithm. We compared the sensitivities of the 2 methods using McNemar's test. A total of 183 of 185 observations (98.9%) demonstrated a Gaussian distribution in adrenal nodule pixel attenuation values. The sensitivity and specificity of our Gaussian model-based algorithm for identifying adrenal adenoma were 86.1% and 83.3%, respectively. The sensitivity and specificity of the mean attenuation method were 53.2% and 94.4%, respectively. The sensitivities of the 2 methods were significantly different (P value < 0.001). In conclusion, the CT attenuation values within an adrenal nodule follow a Gaussian distribution. Our Gaussian model-based algorithm can characterize adrenal adenomas with higher sensitivity than the conventional mean attenuation method. The use of our algorithm, which does not require additional postprocessing, may increase workflow efficiency and reduce unnecessary workup of benign nodules. Copyright © 2016 Elsevier Inc. All rights reserved.

Generation and Characterization of Anti-CD34 Monoclonal Antibodies that React with Hematopoietic Stem Cells

PubMed Central

Aghebati Maleki, Leili; Majidi, Jafar; Baradaran, Behzad; Movassaghpour, Aliakbar; Abdolalizadeh, Jalal

2014-01-01

CD34 is a type I membrane protein with a molecular mass of approximately 110 kDa. This antigen is associated with human hematopoietic progenitor cells and is a differentiation stage-specific leukocyte antigen. In this study we have generated and characterized monoclonal antibodies (mAbs) directed against a CD34 marker. Mice were immunized with two keyhole lympet hemocyanin (KLH)-conjugated CD34 peptides. Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by the limiting dilution (L.D) method. Several monoclones were isolated by three rounds of limited dilutions. From these, we chose stable clones that presented sustained antibody production for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the CD34 peptides and further screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses. One of the mAbs (3D5) was strongly reactive against the CD34 peptide and with native CD34 from human umbilical cord blood cells (UCB) in ELISA and Western blotting analyses. The results have shown that this antibody is highly specific and functional in biomedical applications such as ELISA and Western blot assays. This monoclonal antibodies (mAb) can be a useful tool for isolation and purification of human hematopoietic stem cells (HSCs). PMID:24611141

Novel approaches for an enhanced geothermal development of residential sites

NASA Astrophysics Data System (ADS)

Schelenz, Sophie; Firmbach, Linda; Shao, Haibing; Dietrich, Peter; Vienken, Thomas

2015-04-01

An ongoing technological enhancement drives an increasing use of shallow geothermal systems for heating and cooling applications. However, even in areas with intensive shallow geothermal use, planning of geothermal systems is in many cases solely based on geological maps, drilling databases, and literature references. Thus, relevant heat transport parameters are rather approximated than measured for the specific site. To increase the planning safety and promote the use of renewable energies in the domestic sector, this study investigates a novel concept for an enhanced geothermal development of residential neighbourhoods. This concept is based on a site-specific characterization of subsurface conditions and the implementation of demand-oriented geothermal usage options. Therefore, an investigation approach has been tested that combines non-invasive with minimum-invasive exploration methods. While electrical resistivity tomography has been applied to characterize the geological subsurface structure, Direct Push soundings enable a detailed, vertical high-resolution characterization of the subsurface surrounding the borehole heat exchangers. The benefit of this site-specific subsurface investigation is highlighted for 1) a more precise design of shallow geothermal systems and 2) a reliable prediction of induced long-term changes in groundwater temperatures. To guarantee the financial feasibility and practicability of the novel geothermal development, three different options for its implementation in residential neighbourhoods were consequently deduced.

Introduction to session on materials and structures

NASA Technical Reports Server (NTRS)

Vosteen, L. F.

1978-01-01

A review was given of the development of composites for aircraft. Supporting base technology and the Aircraft Energy Efficiency Composites Program are included. Specific topics discussed include: (1) environmental effects on materials; (2) material quality and chemical characterization; (3) design and analysis methods; (4) structural durability; (5) impact sensitivity; (6) carbon fiber electrical effects; and (7) composite components.

Quantitative assessment of specific defects in roasted ground coffee via infrared-photoacoustic spectroscopy.

PubMed

Dias, Rafael Carlos Eloy; Valderrama, Patrícia; Março, Paulo Henrique; Dos Santos Scholz, Maria Brigida; Edelmann, Michael; Yeretzian, Chahan

2018-07-30

Chemical analyses and sensory evaluation are the most applied methods for quality control of roasted and ground coffee (RG). However, faster alternatives would be highly valuable. Here, we applied infrared-photoacoustic spectroscopy (FTIR-PAS) on RG powder. Mixtures of specific defective beans were blended with healthy (defect-free) Coffea arabica and Coffea canephora bases in specific ratios, forming different classes of blends. Principal Component Analysis allowed predicting the amount/fraction and nature of the defects in blends while partial Least Squares Discriminant Analysis revealed similarities between blends (=samples). A successful predictive model was obtained using six classes of blends. The model could classify 100% of the samples into four classes. The specificities were higher than 0.9. Application of FTIR-PAS on RG coffee to characterize and classify blends has shown to be an accurate, easy, quick and "green" alternative to current methods. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Characterization of Focal Liver Lesions using CEUS and MRI with Liver-Specific Contrast Media: Experience of a Single Radiologic Center.

PubMed

Beyer, Lukas Philipp; Wassermann, Florian; Pregler, Benedikt; Michalik, Katharina; Rennert, Janine; Wiesinger, Isabel; Stroszczynski, Christian; Wiggermann, Philipp; Jung, Ernst Michael

2017-12-01

 The purpose of this study was to compare contrast-enhanced ultrasound (CEUS), magnetic resonance imaging (MRI) using liver-specific contrast agent and a combination of both for the characterization of focal liver lesions (FLL).  83 patients with both benign and malignant liver lesions were examined using CEUS and MRI after the intravenous administration of liver-specific contrast media. All patients had inconclusive results from prior imaging examinations. Histopathological specimens could be obtained in 53 patients. Ultrasound was performed using a multi-frequency curved probe (1 - 6 MHz) after the injection of 1 - 2.4 ml ultrasound contrast media. The sensitivity, specificity, positive predictive value and negative predictive value of CEUS, MRI and a combination of both (CEUS + MRI) were compared.  The sensitivity, specificity, positive and negative predictive values regarding lesion classification were 90.9 %, 70.6 %, 92.3 % and 66.6 %, respectively, for CEUS; 90.9 %, 82.4 %, 95.2 % and 70.0 %, respectively, for MRI; and 96.9 %, 70.6 %, 92.7 % and 85.7 % respectively, for CEUS + MRI. There were no statistically significant differences. 6 malignant lesions were missed using CEUS or MRI alone (false negatives). The use of both modalities combined reduced the false-negative results to 2.  CEUS and MRI with liver-specific contrast media are very reliable and of equal informative value in the characterization of focal liver lesions. The number of false-negative results can be decreased using a combination of the two methods. © Georg Thieme Verlag KG Stuttgart · New York.

The application of NMR and MS methods for detection of adulteration of wine, fruit juices, and olive oil. A review.

PubMed

Ogrinc, N; Kosir, I J; Spangenberg, J E; Kidric, J

2003-06-01

This review covers two important techniques, high resolution nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS), used to characterize food products and detect possible adulteration of wine, fruit juices, and olive oil, all important products of the Mediterranean Basin. Emphasis is placed on the complementary use of SNIF-NMR (site-specific natural isotopic fractionation nuclear magnetic resonance) and IRMS (isotope-ratio mass spectrometry) in association with chemometric methods for detecting the adulteration.

Structural and functional characterization of solute binding proteins for aromatic compounds derived from lignin: p-coumaric acid and related aromatic acids.

PubMed

Tan, Kemin; Chang, Changsoo; Cuff, Marianne; Osipiuk, Jerzy; Landorf, Elizabeth; Mack, Jamey C; Zerbs, Sarah; Joachimiak, Andrzej; Collart, Frank R

2013-10-01

Lignin comprises 15-25% of plant biomass and represents a major environmental carbon source for utilization by soil microorganisms. Access to this energy resource requires the action of fungal and bacterial enzymes to break down the lignin polymer into a complex assortment of aromatic compounds that can be transported into the cells. To improve our understanding of the utilization of lignin by microorganisms, we characterized the molecular properties of solute binding proteins of ATP-binding cassette transporter proteins that interact with these compounds. A combination of functional screens and structural studies characterized the binding specificity of the solute binding proteins for aromatic compounds derived from lignin such as p-coumarate, 3-phenylpropionic acid and compounds with more complex ring substitutions. A ligand screen based on thermal stabilization identified several binding protein clusters that exhibit preferences based on the size or number of aromatic ring substituents. Multiple X-ray crystal structures of protein-ligand complexes for these clusters identified the molecular basis of the binding specificity for the lignin-derived aromatic compounds. The screens and structural data provide new functional assignments for these solute-binding proteins which can be used to infer their transport specificity. This knowledge of the functional roles and molecular binding specificity of these proteins will support the identification of the specific enzymes and regulatory proteins of peripheral pathways that funnel these compounds to central metabolic pathways and will improve the predictive power of sequence-based functional annotation methods for this family of proteins. Copyright © 2013 Wiley Periodicals, Inc.

Structural and functional characterization of solute binding proteins for aromatic compounds derived from lignin: p-coumaric acid and related aromatic acids

PubMed Central

Tan, Kemin; Chang, Changsoo; Cuff, Marianne; Osipiuk, Jerzy; Landorf, Elizabeth; Mack, Jamey C.; Zerbs, Sarah; Joachimiak, Andrzej; Collart, Frank R.

2013-01-01

Lignin comprises 15.25% of plant biomass and represents a major environmental carbon source for utilization by soil microorganisms. Access to this energy resource requires the action of fungal and bacterial enzymes to break down the lignin polymer into a complex assortment of aromatic compounds that can be transported into the cells. To improve our understanding of the utilization of lignin by microorganisms, we characterized the molecular properties of solute binding proteins of ATP.binding cassette transporter proteins that interact with these compounds. A combination of functional screens and structural studies characterized the binding specificity of the solute binding proteins for aromatic compounds derived from lignin such as p-coumarate, 3-phenylpropionic acid and compounds with more complex ring substitutions. A ligand screen based on thermal stabilization identified several binding protein clusters that exhibit preferences based on the size or number of aromatic ring substituents. Multiple X-ray crystal structures of protein-ligand complexes for these clusters identified the molecular basis of the binding specificity for the lignin-derived aromatic compounds. The screens and structural data provide new functional assignments for these solute.binding proteins which can be used to infer their transport specificity. This knowledge of the functional roles and molecular binding specificity of these proteins will support the identification of the specific enzymes and regulatory proteins of peripheral pathways that funnel these compounds to central metabolic pathways and will improve the predictive power of sequence-based functional annotation methods for this family of proteins. PMID:23606130

A Fast and Robust UHPLC-MRM-MS Method to Characterize and Quantify Grape Skin Tannins after Chemical Depolymerization.

PubMed

Pinasseau, Lucie; Verbaere, Arnaud; Roques, Maryline; Meudec, Emmanuelle; Vallverdú-Queralt, Anna; Terrier, Nancy; Boulet, Jean-Claude; Cheynier, Véronique; Sommerer, Nicolas

2016-10-21

A rapid, sensitive, and selective analysis method using ultra high performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UHPLC-QqQ-MS) has been developed for the characterization and quantification of grape skin flavan-3-ols after acid-catalysed depolymerization in the presence of phloroglucinol (phloroglucinolysis). The compound detection being based on specific MS transitions in Multiple Reaction Monitoring (MRM) mode, this fast gradient robust method allows analysis of constitutive units of grape skin proanthocyanidins, including some present in trace amounts, in a single injection, with a throughput of 6 samples per hour. This method was applied to a set of 214 grape skin samples from 107 different red and white grape cultivars grown under two conditions in the vineyard, irrigated or non-irrigated. The results of triplicate analyses confirmed the robustness of the method, which was thus proven to be suitable for high-throughput and large-scale metabolomics studies. Moreover, these preliminary results suggest that analysis of tannin composition is relevant to investigate the genetic bases of grape response to drought.

Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes.

PubMed

Greenough, Lucia; Schermerhorn, Kelly M; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E; Gardner, Andrew F

2016-01-29

Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3'-5' exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Evaluation of smallpox vaccines using variola neutralization

PubMed Central

Damon, Inger K.; Davidson, Whitni B.; Hughes, Christine M.; Olson, Victoria A.; Smith, Scott K.; Holman, Robert C.; Frey, Sharon E.; Newman, Frances; Belshe, Robert B.; Yan, Lihan; Karem, Kevin

2009-01-01

The search for a ‘third’-generation smallpox vaccine has resulted in the development and characterization of several vaccine candidates. A significant barrier to acceptance is the absence of challenge models showing induction of correlates of protective immunity against variola virus. In this light, virus neutralization provides one of few experimental methods to show specific ‘in vitro’ activity of vaccines against variola virus. Here, we provide characterization of the ability of a modified vaccinia virus Ankara vaccine to induce variola virus-neutralizing antibodies, and we provide comparison with the neutralization elicited by standard Dryvax vaccination. PMID:19339477

Mode of action characterization for adverse effect of propranolol in Daphnia magna based on behavior and physiology monitoring and metabolite profiling.

PubMed

Jeong, Tae-Yong; Yoon, Dahye; Kim, Suhkmann; Kim, Hyun Young; Kim, Sang Don

2018-02-01

Studies are underway to gather information about the mode of action (MOA) of emerging pollutants that could guide practical environmental decision making. Previously, we showed that propranolol, an active pharmaceutical ingredient, had adverse effects on Daphnia magna that were similar to its pharmaceutical action. In order to characterize the mode of action of propranolol in D. magna, which is suspected to be organ-specific pharmaceutical action or baseline toxicity, we performed time-series monitoring of behavior along with heart rate measurements and nuclear magnetic resonance (NMR) based metabolite profiling. Principle component analysis (PCA) and hierarchical clustering were used to categorize the mode of action of propranolol among 5 chemicals with different modes of action. The findings showed that the mode of action of propranolol in D. magna is organ-specific and vastly different from those of narcotics, even though metabolite regulation is similar between narcotic and non-narcotic candidates. The method applied in this study seems applicable to rapid characterization of the MOA of other cardiovascular pharmaceutical ingredients. Copyright © 2017 Elsevier Ltd. All rights reserved.

Formation, Physicochemical Characterization, and Thermodynamic Stability of the Amorphous State of Drugs and Excipients.

PubMed

Martino, Piera Di; Magnoni, Federico; Peregrina, Dolores Vargas; Gigliobianco, Maria Rosa; Censi, Roberta; Malaj, Ledjan

2016-01-01

Drugs and excipients used for pharmaceutical applications generally exist in the solid (crystalline or amorphous) state, more rarely as liquid materials. In some cases, according to the physicochemical nature of the molecule, or as a consequence of specific technological processes, a compound may exist exclusively in the amorphous state. In other cases, as a consequence of specific treatments (freezing and spray drying, melting and co-melting, grinding and compression), the crystalline form may convert into a completely or partially amorphous form. An amorphous material shows physical and thermodynamic properties different from the corresponding crystalline form, with profound repercussions on its technological performance and biopharmaceutical properties. Several physicochemical techniques such as X-ray powder diffraction, thermal methods of analysis, spectroscopic techniques, gravimetric techniques, and inverse gas chromatography can be applied to characterize the amorphous form of a compound (drug or excipient), and to evaluate its thermodynamic stability. This review offers a survey of the technologies used to convert a crystalline solid into an amorphous form, and describes the most important techniques for characterizing the amorphous state of compounds of pharmaceutical interest.

Survey and Method for Determination of Trajectory Predictor Requirements

NASA Technical Reports Server (NTRS)

Rentas, Tamika L.; Green, Steven M.; Cate, Karen Tung

2009-01-01

A survey of air-traffic-management researchers, representing a broad range of automation applications, was conducted to document trajectory-predictor requirements for future decision-support systems. Results indicated that the researchers were unable to articulate a basic set of trajectory-prediction requirements for their automation concepts. Survey responses showed the need to establish a process to help developers determine the trajectory-predictor-performance requirements for their concepts. Two methods for determining trajectory-predictor requirements are introduced. A fast-time simulation method is discussed that captures the sensitivity of a concept to the performance of its trajectory-prediction capability. A characterization method is proposed to provide quicker, yet less precise results, based on analysis and simulation to characterize the trajectory-prediction errors associated with key modeling options for a specific concept. Concept developers can then identify the relative sizes of errors associated with key modeling options, and qualitatively determine which options lead to significant errors. The characterization method is demonstrated for a case study involving future airport surface traffic management automation. Of the top four sources of error, results indicated that the error associated with accelerations to and from turn speeds was unacceptable, the error associated with the turn path model was acceptable, and the error associated with taxi-speed estimation was of concern and needed a higher fidelity concept simulation to obtain a more precise result

High specific activity platinum-195m

DOEpatents

Mirzadeh, Saed; Du, Miting; Beets, Arnold L.; Knapp, Jr., Furn F.

2004-10-12

A new composition of matter includes .sup.195m Pt characterized by a specific activity of at least 30 mCi/mg Pt, generally made by method that includes the steps of: exposing .sup.193 Ir to a flux of neutrons sufficient to convert a portion of the .sup.193 Ir to .sup.195m Pt to form an irradiated material; dissolving the irradiated material to form an intermediate solution comprising Ir and Pt; and separating the Pt from the Ir by cation exchange chromatography to produce .sup.195m Pt.

Surveyor assay to diagnose persistent Müllerian duct syndrome in Miniature Schnauzers.

PubMed

Kim, Young June; Kwon, Hyuk Jin; Byun, Hyuk Soo; Yeom, Donguk; Choi, Jea-Hong; Kim, Joong-Hyun; Shim, Hosup

2017-12-31

Persistent Müllerian duct syndrome (PMDS) is a pseudohermaphroditism in males characterized by the presence of Müllerian duct derivatives. As PMDS dogs often lack clinical symptoms, a molecular diagnosis is essential to identify the syndrome in these animals. In this study, a new molecular method using DNA mismatch-specific Surveyor nuclease was developed. The Surveyor nuclease assay identified the AMHR2 mutation that produced PMDS in a Miniature Schnauzer as accurately as that obtained by using the conventional method based on restriction digestion. As an alternative to the current molecular diagnostic method, the new method may result in increased accuracy when detecting PMDS.

Surveyor assay to diagnose persistent Müllerian duct syndrome in Miniature Schnauzers

PubMed Central

Kim, Young June; Kwon, Hyuk Jin; Byun, Hyuk Soo; Yeom, Donguk; Choi, Jea-Hong; Kim, Joong-Hyun

2017-01-01

Persistent Müllerian duct syndrome (PMDS) is a pseudohermaphroditism in males characterized by the presence of Müllerian duct derivatives. As PMDS dogs often lack clinical symptoms, a molecular diagnosis is essential to identify the syndrome in these animals. In this study, a new molecular method using DNA mismatch-specific Surveyor nuclease was developed. The Surveyor nuclease assay identified the AMHR2 mutation that produced PMDS in a Miniature Schnauzer as accurately as that obtained by using the conventional method based on restriction digestion. As an alternative to the current molecular diagnostic method, the new method may result in increased accuracy when detecting PMDS. PMID:27515263

Chemometrics Methods for Specificity, Authenticity and Traceability Analysis of Olive Oils: Principles, Classifications and Applications

PubMed Central

Messai, Habib; Farman, Muhammad; Sarraj-Laabidi, Abir; Hammami-Semmar, Asma; Semmar, Nabil

2016-01-01

Background. Olive oils (OOs) show high chemical variability due to several factors of genetic, environmental and anthropic types. Genetic and environmental factors are responsible for natural compositions and polymorphic diversification resulting in different varietal patterns and phenotypes. Anthropic factors, however, are at the origin of different blends’ preparation leading to normative, labelled or adulterated commercial products. Control of complex OO samples requires their (i) characterization by specific markers; (ii) authentication by fingerprint patterns; and (iii) monitoring by traceability analysis. Methods. These quality control and management aims require the use of several multivariate statistical tools: specificity highlighting requires ordination methods; authentication checking calls for classification and pattern recognition methods; traceability analysis implies the use of network-based approaches able to separate or extract mixed information and memorized signals from complex matrices. Results. This chapter presents a review of different chemometrics methods applied for the control of OO variability from metabolic and physical-chemical measured characteristics. The different chemometrics methods are illustrated by different study cases on monovarietal and blended OO originated from different countries. Conclusion. Chemometrics tools offer multiple ways for quantitative evaluations and qualitative control of complex chemical variability of OO in relation to several intrinsic and extrinsic factors. PMID:28231172

Purity Determination by Capillary Electrophoresis Sodium Hexadecyl Sulfate (CE-SHS): A Novel Application For Therapeutic Protein Characterization.

PubMed

Beckman, Jeff; Song, Yuanli; Gu, Yan; Voronov, Sergey; Chennamsetty, Naresh; Krystek, Stanley; Mussa, Nesredin; Li, Zheng Jian

2018-02-20

Capillary gel electrophoresis using sodium dodecyl sulfate (CE-SDS) is used commercially to provide quantitative purity data for therapeutic protein characterization and release. In CE-SDS, proteins are denatured under reducing or nonreducing conditions in the presence of SDS and electrophoretically separated by molecular weight and hydrodynamic radius through a sieving polymer matrix. Acceptable performance of this method would yield protein peaks that are baseline resolved and symmetrical. Nominal CE-SDS conditions and parameters are not optimal for all therapeutic proteins, specifically for Recombinant Therapeutic Protein-1 (RTP-1), where acceptable resolution and peak symmetry were not achieved. The application of longer alkyl chain detergents in the running buffer matrix substantially improved assay performance. Matrix running buffer containing sodium hexadecyl sulfate (SHS) increased peak resolution and plate count 3- and 8-fold, respectively, compared to a traditional SDS-based running gel matrix. At Bristol-Myers Squibb (BMS), we developed and qualified a viable method for the characterization and release of RTP-1 using an SHS-containing running buffer matrix. This work underscores the potential of detergents other than SDS to enhance the resolution and separation power of CE-based separation methods.

Site-Specific Pyrolysis Induced Cleavage at Aspartic Acid Residue in Peptides and Proteins

PubMed Central

Zhang, Shaofeng; Basile, Franco

2011-01-01

A simple and site-specific non-enzymatic method based on pyrolysis has been developed to cleave peptides and proteins. Pyrolytic cleavage was found to be specific and rapid as it induced a cleavage at the C-terminal side of aspartic acid in the temperature range of 220–250 °C in 10 seconds. Electrospray Ionization (ESI) mass spectrometry (MS) and tandem-MS (MS/MS) were used to characterize and identify pyrolysis cleavage products, confirming that sequence information is conserved after the pyrolysis process in both peptides and protein tested. This suggests that pyrolysis-induced cleavage at aspartyl residues can be used as a rapid protein digestion procedure for the generation of sequence specific protein biomarkers. PMID:17388620

Ultra-high performance supercritical fluid chromatography hyphenated to atmospheric pressure chemical ionization high resolution mass spectrometry for the characterization of fast pyrolysis bio-oils.

PubMed

Crepier, Julien; Le Masle, Agnès; Charon, Nadège; Albrieux, Florian; Duchene, Pascal; Heinisch, Sabine

2018-06-01

Extensive characterization of complex mixtures requires the combination of powerful analytical techniques. A Supercritical Fluid Chromatography (SFC) method was previously developed, for the specific case of fast pyrolysis bio oils, as an alternative to gas chromatography (GC and GC × GC) or liquid chromatography (LC and LC × LC), both separation methods being generally used prior to mass spectrometry (MS) for the characterization of such complex matrices. In this study we investigated the potential of SFC hyphenated to high resolution mass spectrometry (SFC-HRMS) for this characterization using Negative ion Atmospheric Pressure Chemical ionization ((-)APCI) for the ionization source. The interface between SFC and (-)APCI/HRMS was optimized from a mix of model compounds with the objective of maximizing the signal to noise ratio. The main studied parameters included both make-up flow-rate and make-up composition. A methodology for the treatment of APCI/HRMS data is proposed. This latter allowed for the identification of molecular formulae. Both SFC-APCI/HRMS method and data processing method were applied to a mixture of 36 model compounds, first analyzed alone and then spiked in a bio-oil. In both cases, 19 compounds could be detected. Among them 9 could be detected in a fast pyrolysis bio-oil by targeted analysis. The whole procedure was applied to the characterization of a bio-oil using helpful representations such as mass-plots, van Krevelen diagrams and heteroatom class distributions. Finally the results were compared with those obtained with a Fourier Transform ion-cyclotron resonance mass spectrometer (FT-ICR/MS). Copyright © 2018 Elsevier B.V. All rights reserved.

Automatic Keyword Extraction from Individual Documents

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

Rose, Stuart J.; Engel, David W.; Cramer, Nicholas O.

2010-05-03

This paper introduces a novel and domain-independent method for automatically extracting keywords, as sequences of one or more words, from individual documents. We describe the method’s configuration parameters and algorithm, and present an evaluation on a benchmark corpus of technical abstracts. We also present a method for generating lists of stop words for specific corpora and domains, and evaluate its ability to improve keyword extraction on the benchmark corpus. Finally, we apply our method of automatic keyword extraction to a corpus of news articles and define metrics for characterizing the exclusivity, essentiality, and generality of extracted keywords within a corpus.

Apnea Detection Method for Cheyne-Stokes Respiration Analysis on Newborn

NASA Astrophysics Data System (ADS)

Niimi, Taiga; Itoh, Yushi; Natori, Michiya; Aoki, Yoshimitsu

2013-04-01

Cheyne-Stokes respiration is especially prevalent in preterm newborns, but its severity may not be recognized. It is characterized by apnea and cyclical weakening and strengthening of the breathing. We developed a method for detecting apnea and this abnormal respiration and for estimating its malignancy. Apnea was detected based on a "difference" feature (calculated from wavelet coefficients) and a modified maximum displacement feature (related to the respiratory waveform shape). The waveform is calculated from vertical motion of the thoracic and abdominal region during respiration using a vision sensor. Our proposed detection method effectively detects apnea (sensitivity 88.4%, specificity 99.7%).

Specific phobia predicts psychopathology in young women

PubMed Central

Margraf, Jürgen; Vriends, Noortje; Meyer, Andrea H.; Becker, Eni S.

2009-01-01

Background Although specific phobia is characterized by an early age at onset and by high rates of comorbidity, few studies have examined comorbid relationships prospectively. Objectives The present study investigated the association between specific phobia and the risk of a broad range of psychopathology among young women in the community. Method Data came from the Dresden Predictor Study in which 1,538 German women (18–25 years) completed a diagnostic interview at two time points. Results Women with specific phobia had a twofold increase in odds of developing any anxiety disorder, generalized anxiety disorder, depression, and any somatoform disorder during 17 months, compared to women without specific phobia. Except for depression, these associations persisted after adjustment for all comorbid mental disorders. Conclusions Specific phobia thus appears to be a risk factor for a variety of problems. The result further underpins the necessity for early intervention for specific phobia to prevent later mental health problems. PMID:19888542

Minimum Information Specification For In Situ Hybridization and Immunohistochemistry Experiments (MISFISHIE)

PubMed Central

Deutsch, Eric W; Ball, Catherine A; Berman, Jules J; Bova, G Steven; Brazma, Alvis; Bumgarner, Roger E; Campbell, David; Causton, Helen C; Christiansen, Jeffrey H; Daian, Fabrice; Dauga, Delphine; Davidson, Duncan R; Gimenez, Gregory; Goo, Young Ah; Grimmond, Sean; Henrich, Thorsten; Herrmann, Bernhard G; Johnson, Michael H; Korb, Martin; Mills, Jason C; Oudes, Asa J; Parkinson, Helen E; Pascal, Laura E; Pollet, Nicolas; Quackenbush, John; Ramialison, Mirana; Ringwald, Martin; Salgado, David; Sansone, Susanna-Assunta; Sherlock, Gavin; Stoeckert, Christian J; Swedlow, Jason; Taylor, Ronald C; Walashek, Laura; Warford, Anthony; Wilkinson, David G; Zhou, Yi; Zon, Leonard I; Liu, Alvin Y; True, Lawrence D

2015-01-01

One purpose of the biomedical literature is to report results in sufficient detail so that the methods of data collection and analysis can be independently replicated and verified. Here we present for consideration a minimum information specification for gene expression localization experiments, called the “Minimum Information Specification For In Situ Hybridization and Immunohistochemistry Experiments (MISFISHIE)”. It is modelled after the MIAME (Minimum Information About a Microarray Experiment) specification for microarray experiments. Data specifications like MIAME and MISFISHIE specify the information content without dictating a format for encoding that information. The MISFISHIE specification describes six types of information that should be provided for each experiment: Experimental Design, Biomaterials and Treatments, Reporters, Staining, Imaging Data, and Image Characterizations. This specification has benefited the consortium within which it was initially developed and is expected to benefit the wider research community. We welcome feedback from the scientific community to help improve our proposal. PMID:18327244

Technologies and methods used for the detection, enrichment and characterization of cancer stem cells.

PubMed

Williams, Anthony; Datar, Ram; Cote, Richard

2010-01-01

Cancer stem cells (CSCs) represent a subclass of tumour cells with the ability for self-renewal, production of differentiated progeny, prolonged survival, resistance to damaging therapeutic agents, and anchorage-independent survival, which together make this population effectively equipped to metastasize, invade and colonize secondary tissues in the face of therapeutic intervention. In recent years, investigators have increasingly focused on the characterization of CSCs to better understand the mechanisms that govern malignant disease progression in an effort to develop more effective, targeted therapeutic agents. The primary obstacle to the study of CSCs, however, is their rarity. Thus, the study of CSCs requires the use of sensitive and efficient technologies for their enrichment and detection. This review discusses technologies and methods that have been adapted and used to isolate and characterize CSCs to date, as well as new potential directions for the enhanced enrichment and detection of CSCs. While the technologies used for CSC enrichment and detection have been useful thus far for their characterization, each approach is not without limitations. Future studies of CSCs will depend on the enhanced sensitivity and specificity of currently available technologies, and the development of novel technologies for increased detection and enrichment of CSCs.

Rapid Characterization of Microalgae and Microalgae Mixtures Using Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)

PubMed Central

Barbano, Duane; Diaz, Regina; Zhang, Lin; Sandrin, Todd; Gerken, Henri; Dempster, Thomas

2015-01-01

Current molecular methods to characterize microalgae are time-intensive and expensive. Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) may represent a rapid and economical alternative approach. The objectives of this study were to determine whether MALDI-TOF MS can be used to: 1) differentiate microalgae at the species and strain levels and 2) characterize simple microalgal mixtures. A common protein extraction sample preparation method was used to facilitate rapid mass spectrometry-based analysis of 31 microalgae. Each yielded spectra containing between 6 and 56 peaks in the m/z 2,000 to 20,000 range. The taxonomic resolution of this approach appeared higher than that of 18S rDNA sequence analysis. For example, two strains of Scenedesmus acutus differed only by two 18S rDNA nucleotides, but yielded distinct MALDI-TOF mass spectra. Mixtures of two and three microalgae yielded relatively complex spectra that contained peaks associated with members of each mixture. Interestingly, though, mixture-specific peaks were observed at m/z 11,048 and 11,230. Our results suggest that MALDI-TOF MS affords rapid characterization of individual microalgae and simple microalgal mixtures. PMID:26271045

Synthesis and characterization of Co3O4 prepared from atmospheric pressure acid leach liquors of nickel laterite ores

NASA Astrophysics Data System (ADS)

Meng, Long; Guo, Zhan-cheng; Qu, Jing-kui; Qi, Tao; Guo, Qiang; Hou, Gui-hua; Dong, Peng-yu; Xi, Xin-guo

2018-01-01

A chemical precipitation-thermal decomposition method was developed to synthesize Co3O4 nanoparticles using cobalt liquor obtained from the atmospheric pressure acid leaching process of nickel laterite ores. The effects of the precursor reaction temperature, the concentration of Co2+, and the calcination temperature on the specific surface area, morphology, and the electrochemical behavior of the obtained Co3O4 particles were investigated. The precursor basic cobaltous carbonate and cobaltosic oxide products were characterized and analyzed by Fourier transform infrared spectroscopy, thermogravimetric differential thermal analysis, X-ray diffraction, field-emission scanning electron microscopy, specific surface area analysis, and electrochemical analysis. The results indicate that the specific surface area of the Co3O4 particles with a diameter of 30 nm, which were obtained under the optimum conditions of a precursor reaction temperature of 30°C, 0.25 mol/L Co2+, and a calcination temperature of 350°C, was 48.89 m2/g. Electrodes fabricated using Co3O4 nanoparticles exhibited good electrochemical properties, with a specific capacitance of 216.3 F/g at a scan rate of 100 mV/s.

Purification and characterization of VDE, a site-specific endonuclease from the yeast Saccharomyces cerevisiae.

PubMed

Gimble, F S; Thorner, J

1993-10-15

The 119-kDa primary translation product of the VMA1 gene of Saccharomyces cerevisiae undergoes a self-catalyzed rearrangement ("protein splicing") that excises an internal 50-kDa segment of the polypeptide and joins the amino-terminal and carboxyl-terminal segments to generate the 69-kDa subunit of the vacuolar membrane-associated H(+)-ATPase. We have shown previously that the internal segment is a site-specific endonuclease (Gimble, F. S., and Thorner, J. (1992) Nature 357, 301-306). Here we describe methods for the high level expression and purification to near homogeneity of both the authentic VMA1-derived endonuclease (or VDE) from yeast (yield 18%) and a recombinant form of VDE made in bacteria (yield 29%). Detailed characterization of these preparations demonstrated that the yeast-derived and bacterially produced enzymes were indistinguishable, as judged by: (a) behavior during purification; (b) apparent native molecular mass (50 kDa); (c) immunological reactivity; and (d) catalytic properties (specific activity; cleavage site recognition; and optima for pH, temperature, divalent cation and ionic strength). The minimal site required for VDE cleavage was delimited to a 30-base pair sequence within its specific substrate (the VMA1 delta vde allele).

Identification of family-specific residue packing motifs and their use for structure-based protein function prediction: I. Method development.

PubMed

Bandyopadhyay, Deepak; Huan, Jun; Prins, Jan; Snoeyink, Jack; Wang, Wei; Tropsha, Alexander

2009-11-01

Protein function prediction is one of the central problems in computational biology. We present a novel automated protein structure-based function prediction method using libraries of local residue packing patterns that are common to most proteins in a known functional family. Critical to this approach is the representation of a protein structure as a graph where residue vertices (residue name used as a vertex label) are connected by geometrical proximity edges. The approach employs two steps. First, it uses a fast subgraph mining algorithm to find all occurrences of family-specific labeled subgraphs for all well characterized protein structural and functional families. Second, it queries a new structure for occurrences of a set of motifs characteristic of a known family, using a graph index to speed up Ullman's subgraph isomorphism algorithm. The confidence of function inference from structure depends on the number of family-specific motifs found in the query structure compared with their distribution in a large non-redundant database of proteins. This method can assign a new structure to a specific functional family in cases where sequence alignments, sequence patterns, structural superposition and active site templates fail to provide accurate annotation.

A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara.

PubMed

Won, Helen; Yang, Samuel; Gaydos, Charlotte; Hardick, Justin; Ramachandran, Padmini; Hsieh, Yu-Hsiang; Kecojevic, Alexander; Njanpop-Lafourcade, Berthe-Marie; Mueller, Judith E; Tameklo, Tsidi Agbeko; Badziklou, Kossi; Gessner, Bradford D; Rothman, Richard E

2012-09-01

This study aimed to conduct a pilot evaluation of broad-based multiprobe polymerase chain reaction (PCR) in clinical cerebrospinal fluid (CSF) samples compared to local conventional PCR/culture methods used for bacterial meningitis surveillance. A previously described PCR consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by Gram typing, and pathogen-specific probes was designed targeting variable regions of the 16S rRNA gene. The diagnostic performance of the 16S rRNA assay in "127 CSF samples was evaluated in samples from patients from Togo, Africa, by comparison to conventional PCR/culture methods. Our probes detected Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. Uniprobe sensitivity and specificity versus conventional PCR were 100% and 54.6%, respectively. Sensitivity and specificity of uniprobe versus culture methods were 96.5% and 52.5%, respectively. Gram-typing probes correctly typed 98.8% (82/83) and pathogen-specific probes identified 96.4% (80/83) of the positives. This broad-based PCR algorithm successfully detected and provided species level information for multiple bacterial meningitis agents in clinical samples. Copyright © 2012 Elsevier Inc. All rights reserved.

A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara☆, ☆☆,★

PubMed Central

Won, Helen; Yang, Samuel; Gaydos, Charlotte; Hardick, Justin; Ramachandran, Padmini; Hsieh, Yu-Hsiang; Kecojevic, Alexander; Njanpop-Lafourcade, Berthe-Marie; Mueller, Judith E.; Tameklo, Tsidi Agbeko; Badziklou, Kossi; Gessner, Bradford D.; Rothman, Richard E.

2012-01-01

This study aimed to conduct a pilot evaluation of broad-based multiprobe polymerase chain reaction (PCR) in clinical cerebrospinal fluid (CSF) samples compared to local conventional PCR/culture methods used for bacterial meningitis surveillance. A previously described PCR consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by Gram typing, and pathogen-specific probes was designed targeting variable regions of the 16S rRNA gene. The diagnostic performance of the 16S rRNA assay in “”127 CSF samples was evaluated in samples from patients from Togo, Africa, by comparison to conventional PCR/culture methods. Our probes detected Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. Uniprobe sensitivity and specificity versus conventional PCR were 100% and 54.6%, respectively. Sensitivity and specificity of uniprobe versus culture methods were 96.5% and 52.5%, respectively. Gram-typing probes correctly typed 98.8% (82/83) and pathogen-specific probes identified 96.4% (80/83) of the positives. This broad-based PCR algorithm successfully detected and provided species level information for multiple bacterial meningitis agents in clinical samples. PMID:22809694

Characterization of Diffusion Metric Map Similarity in Data From a Clinical Data Repository Using Histogram Distances

PubMed Central

Warner, Graham C.; Helmer, Karl G.

2018-01-01

As the sharing of data is mandated by funding agencies and journals, reuse of data has become more prevalent. It becomes imperative, therefore, to develop methods to characterize the similarity of data. While users can group data based on the acquisition parameters stored in the file headers, these gives no indication whether a file can be combined with other data without increasing the variance in the data set. Methods have been implemented that characterize the signal-to-noise ratio or identify signal drop-outs in the raw image files, but potential users of data often have access to calculated metric maps and these are more difficult to characterize and compare. Here we describe a histogram-distance-based method applied to diffusion metric maps of fractional anisotropy and mean diffusivity that were generated using data extracted from a repository of clinically-acquired MRI data. We describe the generation of the data set, the pitfalls specific to diffusion MRI data, and the results of the histogram distance analysis. We find that, in general, data from GE scanners are less similar than are data from Siemens scanners. We also find that the distribution of distance metric values is not Gaussian at any selection of the acquisition parameters considered here (field strength, number of gradient directions, b-value, and vendor). PMID:29568257

Preparation, Characterization, and Catalytic Activity of MoCo/USY Catalyst on Hydrodeoxygenation Reaction of Anisole

NASA Astrophysics Data System (ADS)

Nugrahaningtyas, K. D.; Suharbiansah, R. S. R.; Rahmawati, F.

2018-03-01

This research aims to prepare, characterize, and study the catalytic activity of Molybdenum (Mo) and Cobalt (Co) metal with supporting material Ultra Stable Y-Zeolite (USY), to produce catalysts with activity in hydrotreatment reaction and in order to eliminate impurities compounds that containing unwanted groups heteroatoms. The bimetallic catalysts MoCo/USY were prepared by wet impregnation method with weight variation of Co metal 0%, 2%, 4%, 6%, 8%, and Mo metal 8% (w/w), respectively. Activation method of the catalyst included calcination, oxidation, reduction and the crystallinity was characterized using X-ray diffraction (XRD), the acidity of the catalyst was analyzed using Fourier Transform Infrared Spectroscopy (FT-IR) and gravimetry method, minerals present in the catalyst was analyzed using X-Ray Fluorescence (XRF), and surface of the catalyst was analyzed using Surface Area Analyzer (SAA). Catalytic activity test (benzene yield product) of MoCo/USY on hydrodeoxigenation reaction of anisole aimed to determine the effect of Mo-Co/USY for catalytic activity in the reaction hydrodeoxigenation (HDO) anisole. Based on characterization and test of catalytic activity, it is known that catalytic of MoCo/USY 2% (catalyst B) shows best activities with acidity of 10.209 mmol/g, specific area of catalyst of 426.295 m2/g, pore average of 14.135 Å, total pore volume 0.318 cc/g, and total yield of HDO products 6.06%.

Identification and characterization of Serpulina hyodysenteriae by restriction enzyme analysis and Southern blot analysis.

PubMed Central

Sotiropoulos, C; Coloe, P J; Smith, S C

1994-01-01

Chromosomal DNA restriction enzyme analysis and Southern blot hybridization were used to characterize Serpulina hyodysenteriae strains. When chromosomal DNAs from selected strains (reference serotypes) of S. hyodysenteriae were digested with the restriction endonuclease Sau3A and hybridized with a 1.1-kb S. hyodysenteriae-specific DNA probe, a common 3-kb band was always detected in S. hyodysenteriae strains but was absent from Serpulina innocens strains. When the chromosomal DNA was digested with the restriction endonuclease Asp 700 and hybridized with two S. hyodysenteriae-specific DNA probes (0.75 and 1.1 kb of DNA), distinct hybridization patterns for each S. hyodysenteriae reference strain and the Australian isolate S. hyodysenteriae 5380 were detected. Neither the 1.1-kb nor the 0.75-kb DNA probe hybridized with Asp 700- or Sau3A-digested S. innocens chromosomal DNA. The presence of the 3-kb Sau3A DNA fragment in S. hyodysenteriae reference strains from diverse geographical locations shows that this fragment is conserved among S. hyodysenteriae strains and can be used as a species-specific marker. Restriction endonuclease analysis and Southern blot hybridization with these well-defined DNA probes are reliable and accurate methods for species-specific and strain-specific identification of S. hyodysenteriae. Images PMID:7914209

Structural Characterization and Absolute Quantification of Microcystin Peptides Using Collision-Induced and Ultraviolet Photo-Dissociation Tandem Mass Spectrometry

NASA Astrophysics Data System (ADS)

Attard, Troy J.; Carter, Melissa D.; Fang, Mengxuan; Johnson, Rudolph C.; Reid, Gavin E.

2018-05-01

Microcystin (MC) peptides produced by cyanobacteria pose a hepatotoxic threat to human health upon ingestion from contaminated drinking water. While rapid MC identification and quantification in contaminated body fluids or tissue samples is important for patient treatment and outcomes, conventional immunoassay-based measurement strategies typically lack the specificity required for unambiguous determination of specific MC variants, whose toxicity can significantly vary depending on their structures. Furthermore, the unambiguous identification and accurate quantitation of MC variants using tandem mass spectrometry (MS/MS)-based methods can be limited due to a current lack of appropriate stable isotope-labeled internal standards. To address these limitations, we have systematically examined here the sequence and charge state dependence to the formation and absolute abundance of both "global" and "variant-specific" product ions from representative MC-LR, MC-YR, MC-RR, and MC-LA peptides, using higher-energy collisional dissociation (HCD)-MS/MS, ion-trap collision-induced dissociation (CID)-MS/MS and CID-MS3, and 193 nm ultraviolet photodissociation (UPVD)-MS/MS. HCD-MS/MS was found to provide the greatest detection sensitivity for both global and variant-specific product ions in each of the MC variants, except for MC-YR where a variant-specific product uniquely formed via UPVD-MS/MS was observed with the greatest absolute abundance. A simple methodology for the preparation and characterization of 18O-stable isotope-labeled MC reference materials for use as internal standards was also developed. Finally, we have demonstrated the applicability of the methods developed herein for absolute quantification of MC-LR present in human urine samples, using capillary scale liquid chromatography coupled with ultra-high resolution / accurate mass spectrometry and HCD-MS/MS.

The simple preparation of birnessite-type manganese oxide with flower-like microsphere morphology and its remarkable capacity retention

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

Zhu, Gang; Deng, Lingjuan; Wang, Jianfang

Graphical abstract: Flower-like birnessite-type manganese oxide microspheres with large specific surface area and excellent electrochemical properties have been prepared by a facile hydrothermal method. Highlights: ► Birnessite-type manganese oxide with flower-like microsphere morphology and large specific surface area. ► A facile low-temperature hydrothermal method. ► Novel flower-like microsphere consists of the thin nano-platelets. ► Birnessite-type manganese oxide exhibits an ideal capacitive behavior and excellent cycling stability. -- Abstract: Birnessite-type manganese oxide with flower-like microsphere morphology and large specific surface area has been prepared by hydrothermal treating a mixture solution of KMnO{sub 4} and (NH{sub 4}){sub 2}SO{sub 4} at 90 °Cmore » for 24 h. The obtained material is characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy and N{sub 2} adsorption–desorption. Results indicate that the birnessite-type manganese oxide shows novel flower-like microsphere morphology and a specific surface area of 280 m{sup 2} g{sup −1}, and the flower-like microsphere consists of the thin nano-platelets. Electrochemical characterization indicates that the prepared material exhibits an ideal capacitive behavior with a capacitance value of 278 F g{sup −1} in 1 mol L{sup −1} Na{sub 2}SO{sub 4} aqueous solution at a scan rate of 5 mV s{sup −1}. Moreover, the prepared manganese oxide electrode shows excellent cycle stability, and the specific capacitance can maintain 98.6% of the initial one after 5000 cycles.« less

Spherical V-Fe-MCM-48: The Synthesis, Characterization and Hydrothermal Stability.

PubMed

Qian, Wang; Wang, Haiqing; Chen, Jin; Kong, Yan

2015-04-14

Spherical MCM-48 mesoporous sieve co-doped with vanadium and iron was successfully synthesized via one-step hydrothermal method. The material was characterized by X-ray diffraction (XRD), nitrogen adsorption-desorption isotherms, inductively coupled plasma (ICP), scanning electron microscopy (SEM), transmission electron microscopy (TEM), diffuse reflectance UV-vis spectra, and X-ray photoelectron spectra (XPS) techniques. Results indicated that the V-Fe-MCM-48 showed an ordered 3D cubic mesostructure with spherical morphology, narrow pore size distribution and high specific surface area. Most of vanadium and iron atoms existing as tetrahedral V 4+ and Fe 3+ species were co-doped into the silicate framework. The particle sizes of V-Fe-MCM-48 were smaller and the specific area was much higher than those of of V-MCM-48. Additionally, the synthesized V-Fe-MCM-48 exhibited improved hydrothermal stability compared with the pure MCM-48.

Spherical V-Fe-MCM-48: The Synthesis, Characterization and Hydrothermal Stability

PubMed Central

Qian, Wang; Wang, Haiqing; Chen, Jin; Kong, Yan

2015-01-01

Spherical MCM-48 mesoporous sieve co-doped with vanadium and iron was successfully synthesized via one-step hydrothermal method. The material was characterized by X-ray diffraction (XRD), nitrogen adsorption-desorption isotherms, inductively coupled plasma (ICP), scanning electron microscopy (SEM), transmission electron microscopy (TEM), diffuse reflectance UV-vis spectra, and X-ray photoelectron spectra (XPS) techniques. Results indicated that the V-Fe-MCM-48 showed an ordered 3D cubic mesostructure with spherical morphology, narrow pore size distribution and high specific surface area. Most of vanadium and iron atoms existing as tetrahedral V4+ and Fe3+ species were co-doped into the silicate framework. The particle sizes of V-Fe-MCM-48 were smaller and the specific area was much higher than those of of V-MCM-48. Additionally, the synthesized V-Fe-MCM-48 exhibited improved hydrothermal stability compared with the pure MCM-48. PMID:28788030

Preparation, Characterization and Application of Optical Switch Probes.

PubMed

Petchprayoon, Chutima; Marriott, Gerard

2010-08-01

Optical switches represent a new class of molecular probe with applications in high contrast imaging and optical manipulation of protein interactions. Small molecule, organic optical switches based on nitrospirobenzopyran (NitroBIPS) and their reactive derivatives and conjugates undergo efficient, rapid and reversible, orthogonal optically-driven transitions between a colorless spiro (SP) state and a colored merocyanine (MC) state. The excited MC-state also emits fluorescence, which serves as readout of the state of the switch. Defined optical perturbations of SP and MC generate a defined waveform of MC-fluorescence that can be isolated against unmodulated background signals by using a digital optical lock-in detection approach or to control specific dipolar interactions on proteins. The protocols describe general procedures for the synthesis and spectroscopic characterization of NitroBIPS and specifically labeled conjugates along with methods for the manipulation of dipolar interactions on proteins and imaging of the MC-state of NitroBIPS within living cells.

Effective material parameter retrieval of anisotropic elastic metamaterials with inherent nonlocality

NASA Astrophysics Data System (ADS)

Lee, Hyung Jin; Lee, Heung Son; Ma, Pyung Sik; Kim, Yoon Young

2016-09-01

In this paper, the scattering (S-) parameter retrieval method is presented specifically for anisotropic elastic metamaterials; so far, no retrieval has been accomplished when elastic metamaterials exhibit fully anisotropic behavior. Complex constitutive property and intrinsic scattering behavior of elastic metamaterials make their characterization far more complicated than that for acoustic and electromagnetic metamaterials. In particular, elastic metamaterials generally exhibit anisotropic scattering behavior due to higher scattering modes associated with shear deformation. They also exhibit nonlocal responses to some degrees, which originate from strong multiple scattering interactions even in the long wavelength limit. Accordingly, the conventional S-parameter retrieval methods cannot be directly used for elastic metamaterials, because they determine only the diagonal components in effective tensor property. Also, the conventional methods simply use the analytic inversion formulae for the material characterization so that inherent nonlocality cannot be taken into account. To establish a retrieval method applicable to anisotropic elastic metamaterials, we propose an alternative S-parameter method to deal with full anisotropy of elastic metamaterials. To retrieve the whole effective anisotropic parameter, we utilize not only normal but also oblique wave incidences. For the retrieval, we first retrieve the ratio of the effective stiffness tensor to effective density and then determine the effective density. The proposed retrieval method is validated by characterizing the effective material parameters of various types of non-resonant anisotropic metamaterials. It is found that the whole effective parameters are retrieved consistently regardless of used retrieval conditions in spite of inherent nonlocality.

Selection, characterization, and application of DNA aptamers for detection of Mycobacterium tuberculosis secreted protein MPT64.

PubMed

Sypabekova, Marzhan; Bekmurzayeva, Aliya; Wang, Ronghui; Li, Yanbin; Nogues, Claude; Kanayeva, Damira

2017-05-01

Rapid detection of Mycobacterium tuberculosis (Mtb), an etiological agent of tuberculosis (TB), is important for global control of this disease. Aptamers have emerged as a potential rival for antibodies in therapeutics, diagnostics and biosensing due to their inherent characteristics. The aim of the current study was to select and characterize single-stranded DNA aptamers against MPT64 protein, one of the predominant secreted proteins of Mtb pathogen. Aptamers specific to MPT64 protein were selected in vitro using systematic evolution of ligands through exponential enrichment (SELEX) method. The selection was started with a pool of ssDNA library with randomized 40-nucleotide region. A total of 10 cycles were performed and seventeen aptamers with unique sequences were identified by sequencing. Dot Blot analysis was performed to monitor the SELEX process and to conduct the preliminary tests on the affinity and specificity of aptamers. Enzyme linked oligonucleotide assay (ELONA) showed that most of the aptamers were specific to the MPT64 protein with a linear correlation of R 2  = 0.94 for the most selective. Using Surface Plasmon Resonance (SPR), dissociation equilibrium constant K D of 8.92 nM was obtained. Bioinformatics analysis of the most specific aptamers revealed the existence of a conserved as well as distinct sequences and possible binding site on MPT64. The specificity was determined by testing non-target ESAT-6 and CFP-10. Negligible cross-reactivity confirmed the high specificity of the selected aptamer. The selected aptamer was further tested on clinical sputum samples using ELONA and had sensitivity and specificity of 91.3% and 90%, respectively. Microscopy, culture positivity and nucleotide amplification methods were used as reference standards. The aptamers studied could be further used for the development of medical diagnostic tools and detection assays for Mtb. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Surveillance methods for identifying, characterizing, and monitoring tobacco products: potential reduced exposure products as an example

PubMed Central

O’Connor, Richard J.; Cummings, K. Michael; Rees, Vaughan W.; Connolly, Gregory N.; Norton, Kaila J.; Sweanor, David; Parascandola, Mark; Hatsukami, Dorothy K.; Shields, Peter G.

2015-01-01

Tobacco products are widely sold and marketed, yet integrated data systems for identifying, tracking, and characterizing products are lacking. Tobacco manufacturers recently have developed potential reduction exposure products (PREPs) with implied or explicit health claims. Currently, a systematic approach for identifying, defining, and evaluating PREPs sold at the local, state or national levels in the US has not been developed. Identifying, characterizing, and monitoring new tobacco products could be greatly enhanced with a responsive surveillance system. This paper critically reviews available surveillance data sources for identifying and tracking tobacco products, including PREPs, evaluating strengths and weaknesses of potential data sources in light of their reliability and validity. Absent regulations mandating disclosure of product-specific information, it is likely that public health officials will need to rely on a variety of imperfect data sources to help identify, characterize, and monitor tobacco products, including PREPs. PMID:19959680

Conformal piezoelectric systems for clinical and experimental characterization of soft tissue biomechanics

NASA Astrophysics Data System (ADS)

Dagdeviren, Canan; Shi, Yan; Joe, Pauline; Ghaffari, Roozbeh; Balooch, Guive; Usgaonkar, Karan; Gur, Onur; Tran, Phat L.; Crosby, Jessi R.; Meyer, Marcin; Su, Yewang; Chad Webb, R.; Tedesco, Andrew S.; Slepian, Marvin J.; Huang, Yonggang; Rogers, John A.

2015-07-01

Mechanical assessment of soft biological tissues and organs has broad relevance in clinical diagnosis and treatment of disease. Existing characterization methods are invasive, lack microscale spatial resolution, and are tailored only for specific regions of the body under quasi-static conditions. Here, we develop conformal and piezoelectric devices that enable in vivo measurements of soft tissue viscoelasticity in the near-surface regions of the epidermis. These systems achieve conformal contact with the underlying complex topography and texture of the targeted skin, as well as other organ surfaces, under both quasi-static and dynamic conditions. Experimental and theoretical characterization of the responses of piezoelectric actuator-sensor pairs laminated on a variety of soft biological tissues and organ systems in animal models provide information on the operation of the devices. Studies on human subjects establish the clinical significance of these devices for rapid and non-invasive characterization of skin mechanical properties.

Microstructural and Defect Characterization in Ceramic Composites Using an Ultrasonic Guided Wave Scan System

NASA Technical Reports Server (NTRS)

Roth, D. J.; Cosgriff, L. M.; Martin, R. E.; Verrilli, M. J.; Bhatt, R. T.

2003-01-01

In this study, an ultrasonic guided wave scan system was used to characterize various microstructural and flaw conditions in two types of ceramic matrix composites, SiC/SiC and C/SiC. Rather than attempting to isolate specific lamb wave modes to use for characterization (as is desired for many types of guided wave inspection problems), the guided wave scan system utilizes the total (multi-mode) ultrasonic response in its inspection analysis. Several time and frequency-domain parameters are calculated from the ultrasonic guided wave signal at each scan location to form images. Microstructural and defect conditions examined include delamination, density variation, cracking, and pre/ post-infiltration. Results are compared with thermographic imaging methods. Although the guided wave technique is commonly used so scanning can be eliminated, applying the technique in the scanning mode allows a more precise characterization of defect conditions.

Allele-specific copy-number discovery from whole-genome and whole-exome sequencing.

PubMed

Wang, WeiBo; Wang, Wei; Sun, Wei; Crowley, James J; Szatkiewicz, Jin P

2015-08-18

Copy-number variants (CNVs) are a major form of genetic variation and a risk factor for various human diseases, so it is crucial to accurately detect and characterize them. It is conceivable that allele-specific reads from high-throughput sequencing data could be leveraged to both enhance CNV detection and produce allele-specific copy number (ASCN) calls. Although statistical methods have been developed to detect CNVs using whole-genome sequence (WGS) and/or whole-exome sequence (WES) data, information from allele-specific read counts has not yet been adequately exploited. In this paper, we develop an integrated method, called AS-GENSENG, which incorporates allele-specific read counts in CNV detection and estimates ASCN using either WGS or WES data. To evaluate the performance of AS-GENSENG, we conducted extensive simulations, generated empirical data using existing WGS and WES data sets and validated predicted CNVs using an independent methodology. We conclude that AS-GENSENG not only predicts accurate ASCN calls but also improves the accuracy of total copy number calls, owing to its unique ability to exploit information from both total and allele-specific read counts while accounting for various experimental biases in sequence data. Our novel, user-friendly and computationally efficient method and a complete analytic protocol is freely available at https://sourceforge.net/projects/asgenseng/. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Frequency and reactivity of antigen-specific T cells were concurrently measured through the combination of artificial antigen-presenting cell, MACS and ELISPOT.

PubMed

Shen, Chuanlai; Xu, Tao; Wu, You; Li, Xiaoe; Xia, Lingzhi; Wang, Wei; Shahzad, Khawar Ali; Zhang, Lei; Wan, Xin; Qiu, Jie

2017-11-27

Conventional peptide-major histocompatibility complex (pMHC) multimer staining, intracellular cytokine staining, and enzyme-linked immunospot (ELISPOT) assay cannot concurrently determine the frequency and reactivity of antigen-specific T cells (AST) in a single assay. In this report, pMHC multimer, magnetic-activated cell sorting (MACS), and ELISPOT techniques have been integrated into a micro well by coupling pMHC multimers onto cell-sized magnetic beads to characterize AST cell populations in a 96-well microplate which pre-coated with cytokine-capture antibodies. This method, termed AAPC-microplate, allows the enumeration and local cytokine production of AST cells in a single assay without using flow cytometry or fluorescence intensity scanning, thus will be widely applicable. Here, ovalbumin 257-264 -specific CD8 + T cells from OT-1 T cell receptor (TCR) transgenic mice were measured. The methodological accuracy, specificity, reproducibility, and sensitivity in enumerating AST cells compared well with conventional pMHC multimer staining. Furthermore, the AAPC-microplate was applied to detect the frequency and reactivity of Hepatitis B virus (HBV) core antigen 18-27 - and surface antigen 183-191 -specific CD8 + T cells for the patients, and was compared with conventional method. This method without the need of high-end instruments may facilitate the routine analysis of patient-specific cellular immune response pattern to a given antigen in translational studies.

Array tomography of physiologically-characterized CNS synapses.

PubMed

Valenzuela, Ricardo A; Micheva, Kristina D; Kiraly, Marianna; Li, Dong; Madison, Daniel V

2016-08-01

The ability to correlate plastic changes in synaptic physiology with changes in synaptic anatomy has been very limited in the central nervous system because of shortcomings in existing methods for recording the activity of specific CNS synapses and then identifying and studying the same individual synapses on an anatomical level. We introduce here a novel approach that combines two existing methods: paired neuron electrophysiological recording and array tomography, allowing for the detailed molecular and anatomical study of synapses with known physiological properties. The complete mapping of a neuronal pair allows determining the exact number of synapses in the pair and their location. We have found that the majority of close appositions between the presynaptic axon and the postsynaptic dendrite in the pair contain synaptic specializations. The average release probability of the synapses between the two neurons in the pair is low, below 0.2, consistent with previous studies of these connections. Other questions, such as receptor distribution within synapses, can be addressed more efficiently by identifying only a subset of synapses using targeted partial reconstructions. In addition, time sensitive events can be captured with fast chemical fixation. Compared to existing methods, the present approach is the only one that can provide detailed molecular and anatomical information of electrophysiologically-characterized individual synapses. This method will allow for addressing specific questions about the properties of identified CNS synapses, even when they are buried within a cloud of millions of other brain circuit elements. Copyright © 2016. Published by Elsevier B.V.

Characterization of sorption properties of selected soils from Lublin region by using water vapour adsorption method

NASA Astrophysics Data System (ADS)

Skic, Kamil; Boguta, Patrycja; Sokołowska, Zofia

2016-04-01

*The studies were carried out within the framework of a research project. The project was financed from funds of National Science Center on the base of decision number DEC-2013/11/D/NZ9/02545 Among many methods proposed to study sorption properties of soils an analysis of adsorption/ desorption isotherm is probably the easiest and most convenient one. It characterizes both quantity and quality of mineral and organic components and also their physical and physicochemical properties. The main aim of this study is comparison of sorption properties of selected Polish soils by using water vapour adsorption method. Samples were taken from the depth of 0-20 cm, from the Lublin region, eastern Poland. Soils were selected on the basis of their different physicochemical properties and were classified as: Haplic Fluvisol, Haplic Chernozem, Mollic Gleysol, Rendzic Phaeozem, Stagnic Luvisol, Haplic Cambisol (WG WRB 2006). Data taken from experimental adsorption isotherms were used to determine parameters of monolayer capacity, specific surface area and the total amount of vapour adsorbed at relative pressure of 0.974. Obtained adsorption and desorption isotherms reviled that adsorbate molecules interacted with the soil particles in different extent. Similar monolayer capacity was observed for Haplic Fluvisol, Haplic Chernozem and Stagnic Luvisol, while for Mollic Gleysol was more than 4 times higher. Mollic Gleysol was also characterized by highest values of specific surface area as well as quantity of adsorbed vapour at relative pressure of 0.974. Higher sorption was caused by presence of soil colloids which contains functional groups of a polar nature (mainly hydroxyls, phenolic and carboxyls). These groups similarly to silicates, oxides, hydratable cations as well as electric charge form adsorption centres for water vapour molecules.

Methodology for cork plank characterization (Quercus suber L.) by near-infrared spectroscopy and image analysis

NASA Astrophysics Data System (ADS)

Prades, Cristina; García-Olmo, Juan; Romero-Prieto, Tomás; García de Ceca, José L.; López-Luque, Rafael

2010-06-01

The procedures used today to characterize cork plank for the manufacture of cork bottle stoppers continue to be based on a traditional, manual method that is highly subjective. Furthermore, there is no specific legislation regarding cork classification. The objective of this viability study is to assess the potential of near-infrared spectroscopy (NIRS) technology for characterizing cork plank according to the following variables: aspect or visual quality, porosity, moisture and geographical origin. In order to calculate the porosity coefficient, an image analysis program was specifically developed in Visual Basic language for a desktop scanner. A set comprising 170 samples from two geographical areas of Andalusia (Spain) was classified into eight quality classes by visual inspection. Spectra were obtained in the transverse and tangential sections of the cork planks using an NIRSystems 6500 SY II reflectance spectrophotometer. The quantitative calibrations showed cross-validation coefficients of determination of 0.47 for visual quality, 0.69 for porosity and 0.66 for moisture. The results obtained using NIRS technology are promising considering the heterogeneity and variability of a natural product such as cork in spite of the fact that the standard error of cross validation (SECV) in the quantitative analysis is greater than the standard error of laboratory (SEL) for the three variables. The qualitative analysis regarding geographical origin achieved very satisfactory results. Applying these methods in industry will permit quality control procedures to be automated, as well as establishing correlations between the different classification systems currently used in the sector. These methods can be implemented in the cork chain of custody certification and will also provide a certainly more objective tool for assessing the economic value of the product.

Comparison Through Image Analysis Between Al Foams Produced Using Two Different Methods

NASA Astrophysics Data System (ADS)

Boschetto, A.; Campana, F.; Pilone, D.

2014-02-01

Several methods are available for making metal foams. They allow to tailor their mechanical, thermal, acoustic, and electrical properties for specific applications by varying the relative density as well as the cell size and morphology. Foams have a very heterogeneous structure so that their properties may show a large scatter. In this paper, an aluminum foam produced by means of foaming of powder compacts and another one prepared via the infiltration process were analyzed and compared. Image analysis has been used as a useful tool to determine size, morphology, and distribution of cells in both foams and to correlate cell morphology with the considered manufacturing process. The results highlighted that cell size and morphology are strictly dependent upon the manufacturing method. This paper shows how some standard 2D morphological indicators may be usefully adopted to characterize foams whose structure derives from the specific manufacturing process.

NetMHCcons: a consensus method for the major histocompatibility complex class I predictions.

PubMed

Karosiene, Edita; Lundegaard, Claus; Lund, Ole; Nielsen, Morten

2012-03-01

A key role in cell-mediated immunity is dedicated to the major histocompatibility complex (MHC) molecules that bind peptides for presentation on the cell surface. Several in silico methods capable of predicting peptide binding to MHC class I have been developed. The accuracy of these methods depends on the data available characterizing the binding specificity of the MHC molecules. It has, moreover, been demonstrated that consensus methods defined as combinations of two or more different methods led to improved prediction accuracy. This plethora of methods makes it very difficult for the non-expert user to choose the most suitable method for predicting binding to a given MHC molecule. In this study, we have therefore made an in-depth analysis of combinations of three state-of-the-art MHC-peptide binding prediction methods (NetMHC, NetMHCpan and PickPocket). We demonstrate that a simple combination of NetMHC and NetMHCpan gives the highest performance when the allele in question is included in the training and is characterized by at least 50 data points with at least ten binders. Otherwise, NetMHCpan is the best predictor. When an allele has not been characterized, the performance depends on the distance to the training data. NetMHCpan has the highest performance when close neighbours are present in the training set, while the combination of NetMHCpan and PickPocket outperforms either of the two methods for alleles with more remote neighbours. The final method, NetMHCcons, is publicly available at www.cbs.dtu.dk/services/NetMHCcons , and allows the user in an automatic manner to obtain the most accurate predictions for any given MHC molecule.

Quantum dot nanoparticle conjugation, characterization, and applications in neuroscience

NASA Astrophysics Data System (ADS)

Pathak, Smita

Quantum dot are semiconducting nanoparticles that have been used for decades in a variety of applications such as solar cells, LEDs and medical imaging. Their use in the last area, however, has been extremely limited despite their potential as revolutionary new biological labeling tools. Quantum dots are much brighter and more stable than conventional fluorophores, making them optimal for high resolution imaging and long term studies. Prior work in this area involves synthesizing and chemically conjugating quantum dots to molecules of interest in-house. However this method is both time consuming and prone to human error. Additionally, non-specific binding and nanoparticle aggregation currently prevent researchers from utilizing this system to its fullest capacity. Another critical issue that has not been addressed is determining the number of ligands bound to nanoparticles, which is crucial for proper interpretation of results. In this work, methods to label fixed cells using two types of chemically modified quantum dots are studied. Reproducible non-specific artifact labeling is consistently demonstrated if antibody-quantum dot conditions are less than optimal. In order to explain this, antibodies bound to quantum dots were characterized and quantified. While other groups have qualitatively characterized antibody functionalized quantum dots using TEM, AFM, UV spectroscopy and gel electrophoresis, and in some cases have reported calculated estimates of the putative number of total antibodies bound to quantum dots, no quantitative experimental results had been reported prior to this work. The chemical functionalization and characterization of quantum dot nanocrystals achieved in this work elucidates binding mechanisms of ligands to nanoparticles and allows researchers to not only translate our tools to studies in their own areas of interest but also derive quantitative results from these studies. This research brings ease of use and increased reliability to nanoparticles in medical imaging.

Thermophysical characterization of the seeds of invasive Chinese tallow tree: importance for biofuel production.

PubMed

Picou, Laura; Boldor, Doran

2012-10-16

The limited supply of traditional fossil based fuels, and increased concern about their environmental impact has driven the interest in the utilization of biomass based energy sources, including those that are underutilized or otherwise nuisance species such as Chinese tallow trees (Triadica sebifera [L.]). This species is a prolific seeds producer, and this paper shows that they contain more than 50% lipids by mass that are suitable for conversion into biodiesel. We present here, for the first time, the seeds' thermophysical properties important for biofuel production. The seeds were characterized using thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and ultimate analysis; their thermal conductivity, thermal diffusivity, and specific heat were determined. The characterization results were correlated to fatty acid composition and lipid content for whole seeds and individual layers, as well as to the protein, hemicellulose, cellulose, and lignin content. The TGA analysis indicated the presence, in addition to lipids, of hemicellulose, cellulose, lignin, and proteins, depending on the layer analyzed. Thermal conductivity and specific heat were, respectively 0.14 ± 0.007 W/mK and 3843.5 ± 171.16 J/kgK for wax, 0.20 ± 0.002 W/mK and 2018.7 ± 5.18 J/kgK for shells, 0.13 ± 0.0 W/mK and 1237 ± 3.15 J/kgK for internal kernel, and 0.13 ± 0.000 W/mK and 2833.9 ± 104.11 J/kgK for whole seeds. These properties and characterization method can be further used in engineering analysis used to determine the most optimum processing method for production of biofuels from this feedstock.

The Role of Extracellular Vesicles in Metastasis

DTIC Science & Technology

2017-10-01

quantitative characterization of each cancerous ESV subpopulation’s role in cargo transfer. Specifically, we aim to (1) optimize an existing...the first quantitative data on which ESV subpopulations (exosomes, MVs, or oncosomes) manipulate the tumor microenvironment, the ESV cargo transferred...without cellular contaminants or without damaging the cargo. A second hindrance has been the lack of quantitative methods for measuring very small

Method calibration of the model 13145 infrared target projectors

NASA Astrophysics Data System (ADS)

Huang, Jianxia; Gao, Yuan; Han, Ying

2014-11-01

The SBIR Model 13145 Infrared Target Projectors ( The following abbreviation Evaluation Unit ) used for characterizing the performances of infrared imaging system. Test items: SiTF, MTF, NETD, MRTD, MDTD, NPS. Infrared target projectors includes two area blackbodies, a 12 position target wheel, all reflective collimator. It provide high spatial frequency differential targets, Precision differential targets imaged by infrared imaging system. And by photoelectricity convert on simulate signal or digital signal. Applications software (IR Windows TM 2001) evaluate characterizing the performances of infrared imaging system. With regards to as a whole calibration, first differently calibration for distributed component , According to calibration specification for area blackbody to calibration area blackbody, by means of to amend error factor to calibration of all reflective collimator, radiance calibration of an infrared target projectors using the SR5000 spectral radiometer, and to analyze systematic error. With regards to as parameter of infrared imaging system, need to integrate evaluation method. According to regulation with -GJB2340-1995 General specification for military thermal imaging sets -testing parameters of infrared imaging system, the results compare with results from Optical Calibration Testing Laboratory . As a goal to real calibration performances of the Evaluation Unit.

Better bioinformatics through usability analysis.

PubMed

Bolchini, Davide; Finkelstein, Anthony; Perrone, Vito; Nagl, Sylvia

2009-02-01

Improving the usability of bioinformatics resources enables researchers to find, interact with, share, compare and manipulate important information more effectively and efficiently. It thus enables researchers to gain improved insights into biological processes with the potential, ultimately, of yielding new scientific results. Usability 'barriers' can pose significant obstacles to a satisfactory user experience and force researchers to spend unnecessary time and effort to complete their tasks. The number of online biological databases available is growing and there is an expanding community of diverse users. In this context there is an increasing need to ensure the highest standards of usability. Using 'state-of-the-art' usability evaluation methods, we have identified and characterized a sample of usability issues potentially relevant to web bioinformatics resources, in general. These specifically concern the design of the navigation and search mechanisms available to the user. The usability issues we have discovered in our substantial case studies are undermining the ability of users to find the information they need in their daily research activities. In addition to characterizing these issues, specific recommendations for improvements are proposed leveraging proven practices from web and usability engineering. The methods and approach we exemplify can be readily adopted by the developers of bioinformatics resources.

Development, Validation, and Application of a Novel Ligand-Binding Assay to Selectively Measure PEGylated Recombinant Human Coagulation Factor VIII (BAX 855).

PubMed

Weber, Alfred; Engelmaier, Andrea; Hainzelmayer, Sandra; Minibeck, Eva; Anderle, Heinz; Schwarz, Hans Peter; Turecek, Peter L

2015-10-21

BAX 855 is a PEGylated recombinant factor VIII preparation that showed prolonged circulatory half-life in nonclinical and clinical studies. This paper describes the development, validation, and application of a novel ligand-binding assay (LBA) to selectively measure BAX 855 in plasma. The LBA is based on PEG-specific capture of BAX 855, followed by immunological factor VIII (FVIII)-specific detection of the antibody-bound BAX 855. This assay principle enabled sensitive measurement of BAX 855 down to the low nanomolar range without interference from non-PEGylated FVIII as demonstrated by validation data for plasma from animals typically used for nonclinical characterization of FVIII. The selectivity of an in-house-developed anti-PEG and a commercially available preparation, shown by competition studies to primarily target the terminating methoxy group of PEG, also allowed assessment of the intactness of the attached PEG chains. Altogether, this new LBA adds to the group of methods to selectively, accurately, and precisely measure a PEGylated drug in complex biological matrices. The feasibility and convenience of using this method was demonstrated during extensive nonclinical characterization of BAX 855.

A Lagrangian cylindrical coordinate system for characterizing dynamic surface geometry of tubular anatomic structures.

PubMed

Lundh, Torbjörn; Suh, Ga-Young; DiGiacomo, Phillip; Cheng, Christopher

2018-03-03

Vascular morphology characterization is useful for disease diagnosis, risk stratification, treatment planning, and prediction of treatment durability. To quantify the dynamic surface geometry of tubular-shaped anatomic structures, we propose a simple, rigorous Lagrangian cylindrical coordinate system to monitor well-defined surface points. Specifically, the proposed system enables quantification of surface curvature and cross-sectional eccentricity. Using idealized software phantom examples, we validate the method's ability to accurately quantify longitudinal and circumferential surface curvature, as well as eccentricity and orientation of eccentricity. We then apply the method to several medical imaging data sets of human vascular structures to exemplify the utility of this coordinate system for analyzing morphology and dynamic geometric changes in blood vessels throughout the body. Graphical abstract Pointwise longitudinal curvature of a thoracic aortic endograft surface for systole and diastole, with their absolute difference.

Laboratory analytical methods for the determination of the hydrocarbon status of soils (a review)

NASA Astrophysics Data System (ADS)

Pikovskii, Yu. I.; Korotkov, L. A.; Smirnova, M. A.; Kovach, R. G.

2017-10-01

Laboratory analytical methods suitable for the determination of the hydrocarbon status of soils (a specific soil characteristic involving information on the total content and qualitative features of soluble (bitumoid) carbonaceous substances and individual hydrocarbons (polycyclic aromatic hydrocarbons, alkanes, etc.) in bitumoid, as well as the composition and content of hydrocarbon gases) have been considered. Among different physicochemical methods of study, attention is focused on the methods suitable for the wide use. Luminescence-bituminological analysis, low-temperature spectrofluorimetry (Shpolskii spectroscopy), infrared (IR) spectroscopy, gas chromatography, chromatography-mass spectrometry, and some other methods have been characterized, as well as sample preparation features. Advantages and limitations of each of these methods are described; their efficiency, instrumental complexity, analysis duration, and accuracy are assessed.

Optimization and Validation of a Plaque Reduction Neutralization Test for the Detection of Neutralizing Antibodies to Four Serotypes of Dengue Virus Used in Support of Dengue Vaccine Development

PubMed Central

Timiryasova, Tatyana M.; Bonaparte, Matthew I.; Luo, Ping; Zedar, Rebecca; Hu, Branda T.; Hildreth, Stephen W.

2013-01-01

A dengue plaque reduction neutralization test (PRNT) to measure dengue serotype–specific neutralizing antibodies for all four virus serotypes was developed, optimized, and validated in accordance with guidelines for validation of bioanalytical test methods using human serum samples from dengue-infected persons and persons receiving a dengue vaccine candidate. Production and characterization of dengue challenge viruses used in the assay was standardized. Once virus stocks were characterized, the dengue PRNT50 for each of the four serotypes was optimized according to a factorial design of experiments approach for critical test parameters, including days of cell seeding before testing, percentage of overlay carboxymethylcellulose medium, and days of incubation post-infection to generate a robust assay. The PRNT50 was then validated and demonstrated to be suitable to detect and measure dengue serotype-specific neutralizing antibodies in human serum samples with acceptable intra-assay and inter-assay precision, accuracy/dilutability, specificity, and with a lower limit of quantitation of 10. PMID:23458954

[Characterization of puroindolines in the control of endosperm texture in common wheat lines with substitutions of homeologous group-5 chromosomes].

PubMed

Obukhova, L V; Efremova, T T; Shumnyĭ, V K

2013-03-01

The genetic control of grain hardness and its association with the specific friabilin content on starch granules of common wheat cultivars and lines with intervarietal substitutions of homeologous group-5 chromosomes were studied. A significant correlation was revealed between the technological parameters of grain hardness (mean size of flour particles) and the specific content of puroindolines on the starch surface estimated in terms of starch doses. The results obtained allowed the method of starch doses to be used to identify soft and hard wheat cultivars and lines based on an analysis of a single grain. The biochemical analysis confirmed the previously obtained estimates of flour-grinding properties of wheat cultivars and substitution lines, which allowed specific genotypes to be characterized according to the composition of puroindolines. The influence of chromosomes 5D and 5A of donor wheat cultivars on the activity of the Ha loci of recipient cultivars was revealed and found to be associated with the composition of PIN products and with the expression of the Pina-D1 and Pinb-D1 genes.

Autoradiographic analysis of binding sites for sup 125 I-Bolton-Hunter-substance P in the human eye

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

Kieselbach, G.F.; Ragaut, R.; Knaus, H.G.

1990-07-01

Substance P is known to exert potent effects in peripheral tissues, and is thought to be important for ocular function. The mechanism of action of substance P in the human eye is not known. As a basis for biochemical characterization specific binding of {sup 125}I-Bolton-Hunter-substance P was demonstrated in the human eye using autoradiographic methods. Biochemical characterization on slide-mounted tissue preparations showed that binding was saturable with a KD of 0.27 +/- 0.1 nmol/l. Specific binding occurred at comparable autoradiographic densities to both human retina and choroid. Substance P and its carboxyterminal fragment, substance P(3-11), were shown to be highlymore » potent in binding competition experiments against {sup 125}I-Bolton-Hunter-substance P. Similar concentrations of substance P(1-9), neurokinin A and neurokinin B failed to significantly alter specific binding of {sup 125}I-Bolton-Hunter-substance P. The results indicate expression of high affinity substance P binding sites in human retina and choroid.« less

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

NASA Astrophysics Data System (ADS)

Zhukhlistova, N. E.; Balaev, V. V.; Lyashenko, A. V.; Lashkov, A. A.

2012-05-01

Endonucleases (EC 3.1) are enzymes of the hydrolase class that catalyze the hydrolytic cleavage of deoxyribonucleic and ribonucleic acids at any region of the polynucleotide chain. Endonucleases are widely used both in biotechnological processes and in veterinary medicine as antiviral agents. Medical applications of endonucleases in human cancer therapy hold promise. The results of X-ray diffraction studies of the spatial organization of endonucleases and their complexes and the mechanism of their action are analyzed and generalized. An analysis of the structural studies of this class of enzymes showed that the specific binding of enzymes to nucleic acids is characterized by interactions with nitrogen bases and the nucleotide backbone, whereas the nonspecific binding of enzymes is generally characterized by interactions only with the nucleic-acid backbone. It should be taken into account that the specificity can be modulated by metal ions and certain low-molecular-weight organic compounds. To test the hypotheses about specific and nonspecific nucleic-acid-binding proteins, it is necessary to perform additional studies of atomic-resolution three-dimensional structures of enzyme-nucleic-acid complexes by methods of structural biology.

Ratiometric spectral imaging for fast tumor detection and chemotherapy monitoring in vivo

PubMed Central

Hwang, Jae Youn; Gross, Zeev; Gray, Harry B.; Medina-Kauwe, Lali K.; Farkas, Daniel L.

2011-01-01

We report a novel in vivo spectral imaging approach to cancer detection and chemotherapy assessment. We describe and characterize a ratiometric spectral imaging and analysis method and evaluate its performance for tumor detection and delineation by quantitatively monitoring the specific accumulation of targeted gallium corrole (HerGa) into HER2-positive (HER2 +) breast tumors. HerGa temporal accumulation in nude mice bearing HER2 + breast tumors was monitored comparatively by a. this new ratiometric imaging and analysis method; b. established (reflectance and fluorescence) spectral imaging; c. more commonly used fluorescence intensity imaging. We also tested the feasibility of HerGa imaging in vivo using the ratiometric spectral imaging method for tumor detection and delineation. Our results show that the new method not only provides better quantitative information than typical spectral imaging, but also better specificity than standard fluorescence intensity imaging, thus allowing enhanced in vivo outlining of tumors and dynamic, quantitative monitoring of targeted chemotherapy agent accumulation into them. PMID:21721808

Tracing the source of difficult to settle fine particles which cause turbidity in the Hitotsuse Reservoir, Japan.

PubMed

Murakami, Toshiki; Suzuki, Yoshihiro; Oishi, Hiroyuki; Ito, Kenichi; Nakao, Toshio

2013-05-15

A unique method to trace the source of "difficult-to-settle fine particles," which are a causative factor of long-term turbidity in reservoirs was developed. This method is characterized by cluster analysis of XRD (X-ray diffraction) data and homology comparison of major component compositions between "difficult-to-settle fine particles" contained in landslide soil samples taken from the upstream of a dam, and suspended "long-term turbid water particles" in the reservoir, which is subject to long-term turbidity. The experiment carried out to validate the proposed method, demonstrated a high possibility of being able to make an almost identical match between "difficult-to-settle fine particles" taken from landslide soils at specific locations and "long-term turbid water particles" taken from a reservoir. This method has the potential to determine substances causing long-term turbidity and the locations of soils from which those substances came. Appropriate countermeasures can then be taken at those specific locations. Copyright © 2013 Elsevier Ltd. All rights reserved.

Will isomalto-oligosaccharides, a well-established functional food in Asia, break through the European and American market? The status of knowledge on these prebiotics.

PubMed

Goffin, Dorothee; Delzenne, Nathalie; Blecker, Christophe; Hanon, Emilien; Deroanne, Claude; Paquot, Michel

2011-05-01

This critical review article presents the current state of knowledge on isomalto-oligosaccharides, some well known functional oligosaccharides in Asia, to evaluate their potential as emergent prebiotics in the American and European functional food market. It includes first a unique inventory of the different families of compounds which have been considered as IMOs and their specific structure. A description has been given of the different production methods including the involved enzymes and their specific activities, the substrates, and the types of IMOs produced. Considering the structural complexity of IMO products, specific characterization methods are described, as well as purification methods which enable the body to get rid of digestible oligosaccharides. Finally, an extensive review of their techno-functional and nutritional properties enables placing IMOs inside the growing prebiotic market. This review is of particular interest considering that IMO commercialization in America and Europe is a topical subject due to the recent submission by Bioneutra Inc. (Canada) of a novel food file to the UK Food Standards Agency, as well as several patents for IMO production.

Bentonite modification with pillarization method using metal stannum

NASA Astrophysics Data System (ADS)

Widjaya, Robert R.; Juwono, Ariadne L.; Rinaldi, Nino

2017-11-01

Clay minerals have received considerable attention in the last years because of their environmental compatibility, low cost, high selectivity, and operational simplicity. Although clays are very useful for many application in the field of catalysis, they have main disadvantage: their lack of pore volume and spesific surface area. Porosity and stability of these materials are improved by pillaring the clay layers with SnCl4, which leads to materials known as pillared clays (PILC). This research aims were to characterize the Bentonite and Sn-Bentonite as catalysts for cracking and oligomerization. The Sn-Bentonite was prepared by pillarization method with a variation in metal ratio of 5 mmol dan 10 mmol.gr-1 of bentonit. The catalyst characterized by X-ray Diffraction, X-ray Fluorescence, Fourier Transform Infra Red, Brunauer Emmett Teller, Thermogravimetric Analysis. The results showed that the Sn-Bentonite catalyst had large basal spacing and good porous structure, and the specific surface areas increased. XRF detected the Sn in the Bentonite and TGA results showed the ability Sn-Bentonite in receiving heat. FTIR test showed two type of acidity, broansted and lewis acid. The characterized results indicated that Sn-Bentonite with metal ratio 5 mmol.gr-1 better than Sn-Bentonite with metal ratio 10 mmol.gr-1, in which there was a significant increase the basal spacings, specific surface area, and pore volume. The TGA results for Sn-Bentonite appeared to be more thermally stable than Bentonite.

Poliovirus serotype-specific VP1 sequencing primers.

PubMed

Kilpatrick, David R; Iber, Jane C; Chen, Qi; Ching, Karen; Yang, Su-Ju; De, Lina; Mandelbaum, Mark D; Emery, Brian; Campagnoli, Ray; Burns, Cara C; Kew, Olen

2011-06-01

The Global Polio Laboratory Network routinely uses poliovirus-specific PCR primers and probes to determine the serotype and genotype of poliovirus isolates obtained as part of global poliovirus surveillance. To provide detailed molecular epidemiologic information, poliovirus isolates are further characterized by sequencing the ~900-nucleotide region encoding the major capsid protein, VP1. It is difficult to obtain quality sequence information when clinical or environmental samples contain poliovirus mixtures. As an alternative to conventional methods for resolving poliovirus mixtures, sets of serotype-specific primers were developed for amplifying and sequencing the VP1 regions of individual components of mixed populations of vaccine-vaccine, vaccine-wild, and wild-wild polioviruses. Published by Elsevier B.V.

A Method to Quantify Visual Information Processing in Children Using Eye Tracking

PubMed Central

Kooiker, Marlou J.G.; Pel, Johan J.M.; van der Steen-Kant, Sanny P.; van der Steen, Johannes

2016-01-01

Visual problems that occur early in life can have major impact on a child's development. Without verbal communication and only based on observational methods, it is difficult to make a quantitative assessment of a child's visual problems. This limits accurate diagnostics in children under the age of 4 years and in children with intellectual disabilities. Here we describe a quantitative method that overcomes these problems. The method uses a remote eye tracker and a four choice preferential looking paradigm to measure eye movement responses to different visual stimuli. The child sits without head support in front of a monitor with integrated infrared cameras. In one of four monitor quadrants a visual stimulus is presented. Each stimulus has a specific visual modality with respect to the background, e.g., form, motion, contrast or color. From the reflexive eye movement responses to these specific visual modalities, output parameters such as reaction times, fixation accuracy and fixation duration are calculated to quantify a child's viewing behavior. With this approach, the quality of visual information processing can be assessed without the use of communication. By comparing results with reference values obtained in typically developing children from 0-12 years, the method provides a characterization of visual information processing in visually impaired children. The quantitative information provided by this method can be advantageous for the field of clinical visual assessment and rehabilitation in multiple ways. The parameter values provide a good basis to: (i) characterize early visual capacities and consequently to enable early interventions; (ii) compare risk groups and follow visual development over time; and (iii), construct an individual visual profile for each child. PMID:27500922

A Method to Quantify Visual Information Processing in Children Using Eye Tracking.

PubMed

Kooiker, Marlou J G; Pel, Johan J M; van der Steen-Kant, Sanny P; van der Steen, Johannes

2016-07-09

Visual problems that occur early in life can have major impact on a child's development. Without verbal communication and only based on observational methods, it is difficult to make a quantitative assessment of a child's visual problems. This limits accurate diagnostics in children under the age of 4 years and in children with intellectual disabilities. Here we describe a quantitative method that overcomes these problems. The method uses a remote eye tracker and a four choice preferential looking paradigm to measure eye movement responses to different visual stimuli. The child sits without head support in front of a monitor with integrated infrared cameras. In one of four monitor quadrants a visual stimulus is presented. Each stimulus has a specific visual modality with respect to the background, e.g., form, motion, contrast or color. From the reflexive eye movement responses to these specific visual modalities, output parameters such as reaction times, fixation accuracy and fixation duration are calculated to quantify a child's viewing behavior. With this approach, the quality of visual information processing can be assessed without the use of communication. By comparing results with reference values obtained in typically developing children from 0-12 years, the method provides a characterization of visual information processing in visually impaired children. The quantitative information provided by this method can be advantageous for the field of clinical visual assessment and rehabilitation in multiple ways. The parameter values provide a good basis to: (i) characterize early visual capacities and consequently to enable early interventions; (ii) compare risk groups and follow visual development over time; and (iii), construct an individual visual profile for each child.

Spectral Induced Polarization approaches to characterize reactive transport parameters and processes

NASA Astrophysics Data System (ADS)

Schmutz, M.; Franceschi, M.; Revil, A.; Peruzzo, L.; Maury, T.; Vaudelet, P.; Ghorbani, A.; Hubbard, S. S.

2017-12-01

For almost a decade, geophysical methods have explored the potential for characterization of reactive transport parameters and processes relevant to hydrogeology, contaminant remediation, and oil and gas applications. Spectral Induced Polarization (SIP) methods show particular promise in this endeavour, given the sensitivity of the SIP signature to geological material electrical double layer properties and the critical role of the electrical double layer on reactive transport processes, such as adsorption. In this presentation, we discuss results from several recent studies that have been performed to quantify the value of SIP parameters for characterizing reactive transport parameters. The advances have been realized through performing experimental studies and interpreting their responses using theoretical and numerical approaches. We describe a series of controlled experimental studies that have been performed to quantify the SIP responses to variations in grain size and specific surface area, pore fluid geochemistry, and other factors. We also model chemical reactions at the interface fluid/matrix linked to part of our experimental data set. For some examples, both geochemical modelling and measurements are integrated into a SIP physico-chemical based model. Our studies indicate both the potential of and the opportunity for using SIP to estimate reactive transport parameters. In case of well sorted granulometry of the samples, we find that the grain size characterization (as well as the permeabililty for some specific examples) value can be estimated using SIP. We show that SIP is sensitive to physico-chemical conditions at the fluid/mineral interface, including the different pore fluid dissolved ions (Na+, Cu2+, Zn2+, Pb2+) due to their different adsorption behavior. We also showed the relevance of our approach to characterize the fluid/matrix interaction for various organic contents (wetting and non-wetting oils). We also discuss early efforts to jointly interpret SIP and other information for improved estimation, approaches to use SIP information to constrain mechanistic flow and transport models, and the potential to apply some of the approaches to field scale applications.

Characterizing lentic freshwater fish assemblages using multiple sampling methods

USGS Publications Warehouse

Fischer, Jesse R.; Quist, Michael C.

2014-01-01

Characterizing fish assemblages in lentic ecosystems is difficult, and multiple sampling methods are almost always necessary to gain reliable estimates of indices such as species richness. However, most research focused on lentic fish sampling methodology has targeted recreationally important species, and little to no information is available regarding the influence of multiple methods and timing (i.e., temporal variation) on characterizing entire fish assemblages. Therefore, six lakes and impoundments (48–1,557 ha surface area) were sampled seasonally with seven gear types to evaluate the combined influence of sampling methods and timing on the number of species and individuals sampled. Probabilities of detection for species indicated strong selectivities and seasonal trends that provide guidance on optimal seasons to use gears when targeting multiple species. The evaluation of species richness and number of individuals sampled using multiple gear combinations demonstrated that appreciable benefits over relatively few gears (e.g., to four) used in optimal seasons were not present. Specifically, over 90 % of the species encountered with all gear types and season combinations (N = 19) from six lakes and reservoirs were sampled with nighttime boat electrofishing in the fall and benthic trawling, modified-fyke, and mini-fyke netting during the summer. Our results indicated that the characterization of lentic fish assemblages was highly influenced by the selection of sampling gears and seasons, but did not appear to be influenced by waterbody type (i.e., natural lake, impoundment). The standardization of data collected with multiple methods and seasons to account for bias is imperative to monitoring of lentic ecosystems and will provide researchers with increased reliability in their interpretations and decisions made using information on lentic fish assemblages.

Identification of Trypanosoma cruzi discrete typing units (DTUs) through the implementation of a high-resolution melting (HRM) genotyping assay.

PubMed

Higuera, Sonia L; Guhl, Felipe; Ramírez, Juan David

2013-04-20

Chagas disease, caused by Trypanosoma cruzi, is a geographically widespread anthropozoonosis that is considered a major public health problem in Latin America. Because this parasite presents high genetic variability, a nomenclature has been adopted to classify the parasite into six discrete typing units (DTUs): TcI, TcII, TcIII, TcIV, TcV, and TcVI, which present different eco-epidemiological, clinical, and geographic associations. Currently, the available genotyping methods present a series of drawbacks that implies the need for developing new methods for characterizing T. cruzi DTU's. The aim of this work was to genotype reference populations from T. cruzi by means of a High-Resolution Melting (HRM) genotyping assay. Amplification of the mini-exon gene allowed the genotyping of three distinct groups: TcI, TcII- TcIV- TcV, and TcIII-TcVI, while amplification of the 24sα gene generated non-overlapping melting temperature ranges for each DTU that were used to identify the groups in the six existing DTUs of Trypanosoma cruzi. The proposed genotyping assay allowed discrimination of the six genetic groups by obtaining specific melting curves for each DTU. The application of this technique is proposed because of its specificity, sensitivity, high performance, and low cost compared with other previously described characterization methods.

Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients

PubMed Central

Livério, Harisson Oliveira; Ruiz, Luciana da Silva; de Freitas, Roseli Santos; Nishikaku, Angela; de Souza, Ana Clara; Paula, Claudete Rodrigues; Domaneschi, Carina

2017-01-01

ABSTRACT The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species. PMID:28423089

Twisting short dsDNA with applied tension

NASA Astrophysics Data System (ADS)

Zoli, Marco

2018-02-01

The twisting deformation of mechanically stretched DNA molecules is studied by a coarse grained Hamiltonian model incorporating the fundamental interactions that stabilize the double helix and accounting for the radial and angular base pair fluctuations. The latter are all the more important at short length scales in which DNA fragments maintain an intrinsic flexibility. The presented computational method simulates a broad ensemble of possible molecule conformations characterized by a specific average twist and determines the energetically most convenient helical twist by free energy minimization. As this is done for any external load, the method yields the characteristic twist-stretch profile of the molecule and also computes the changes in the macroscopic helix parameters i.e. average diameter and rise distance. It is predicted that short molecules under stretching should first over-twist and then untwist by increasing the external load. Moreover, applying a constant load and simulating a torsional strain which over-twists the helix, it is found that the average helix diameter shrinks while the molecule elongates, in agreement with the experimental trend observed in kilo-base long sequences. The quantitative relation between percent relative elongation and superhelical density at fixed load is derived. The proposed theoretical model and computational method offer a general approach to characterize specific DNA fragments and predict their macroscopic elastic response as a function of the effective potential parameters of the mesoscopic Hamiltonian.

Mass-Production and Characterization of Anti-CD20 Monoclonal Antibody in Peritoneum of Balb/c Mice

PubMed Central

Sineh sepehr, Koushan; Baradaran, Behzad; Majidi, Jafar; Abdolalizadeh, Jalal; Aghebati, leili; Zare Shahneh, Fatemeh

2013-01-01

Purpose: Monoclonal antibodies are important tools are used in basic research as well as, in diagnosis, imaging and treatment of immunodeficiency diseases, infections and cancers. The purpose of this study was to produce large scale of monoclonal antibody against CD20 in order to diagnostic application in leukemia and lymphomas disorders. Methods: Hybridoma cells that produce monoclonal antibody against human CD20 were administered into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. After twelve days, approximately 7 ml ascetic fluid was harvested from the peritoneum of each mouse. Evaluation of mAb titration was assessed by ELISA method. In the present study, we describe a protocol for large scale production of MAbs. Results: We prepared monoclonal antibodies (mAbs) with high specificity and sensitivity against human CD20 by hybridoma method and characterized them by ELISA. The subclass of antibody was IgG2a and its light chain was kappa. Ascetic fluid was purified by Protein-A Sepharose affinity chromatography and the purified monoclonal antibody was conjugated with FITC and Immunofluorescence was done for confirming the specific binding. Conclusion: The conjugated monoclonal antibody could have application in diagnosis B-cell lymphomas, hairy cell leukemia, B-cell chronic lymphocytic leukemia, and melanoma cancer stem cells. PMID:24312821

Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients.

PubMed

Livério, Harisson Oliveira; Ruiz, Luciana da Silva; Freitas, Roseli Santos de; Nishikaku, Angela; Souza, Ana Clara de; Paula, Claudete Rodrigues; Domaneschi, Carina

2017-04-13

The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species.

Manipulation-free cultures of human iPSC-derived cardiomyocytes offer a novel screening method for cardiotoxicity.

PubMed

Rajasingh, Sheeja; Isai, Dona Greta; Samanta, Saheli; Zhou, Zhi-Gang; Dawn, Buddhadeb; Kinsey, William H; Czirok, Andras; Rajasingh, Johnson

2018-04-05

Induced pluripotent stem cell (iPSC)-based cardiac regenerative medicine requires the efficient generation, structural soundness and proper functioning of mature cardiomyocytes, derived from the patient's somatic cells. The most important functional property of cardiomyocytes is the ability to contract. Currently available methods routinely used to test and quantify cardiomyocyte function involve techniques that are labor-intensive, invasive, require sophisticated instruments or can adversely affect cell vitality. We recently developed optical flow imaging method analyses and quantified cardiomyocyte contractile kinetics from video microscopic recordings without compromising cell quality. Specifically, our automated particle image velocimetry (PIV) analysis of phase-contrast video images captured at a high frame rate yields statistical measures characterizing the beating frequency, amplitude, average waveform and beat-to-beat variations. Thus, it can be a powerful assessment tool to monitor cardiomyocyte quality and maturity. Here we demonstrate the ability of our analysis to characterize the chronotropic responses of human iPSC-derived cardiomyocytes to a panel of ion channel modulators and also to doxorubicin, a chemotherapy agent with known cardiotoxic side effects. We conclude that the PIV-derived beat patterns can identify the elongation or shortening of specific phases in the contractility cycle, and the obtained chronotropic responses are in accord with known clinical outcomes. Hence, this system can serve as a powerful tool to screen the new and currently available pharmacological compounds for cardiotoxic effects.

A grammar inference approach for predicting kinase specific phosphorylation sites.

PubMed

Datta, Sutapa; Mukhopadhyay, Subhasis

2015-01-01

Kinase mediated phosphorylation site detection is the key mechanism of post translational mechanism that plays an important role in regulating various cellular processes and phenotypes. Many diseases, like cancer are related with the signaling defects which are associated with protein phosphorylation. Characterizing the protein kinases and their substrates enhances our ability to understand the mechanism of protein phosphorylation and extends our knowledge of signaling network; thereby helping us to treat such diseases. Experimental methods for predicting phosphorylation sites are labour intensive and expensive. Also, manifold increase of protein sequences in the databanks over the years necessitates the improvement of high speed and accurate computational methods for predicting phosphorylation sites in protein sequences. Till date, a number of computational methods have been proposed by various researchers in predicting phosphorylation sites, but there remains much scope of improvement. In this communication, we present a simple and novel method based on Grammatical Inference (GI) approach to automate the prediction of kinase specific phosphorylation sites. In this regard, we have used a popular GI algorithm Alergia to infer Deterministic Stochastic Finite State Automata (DSFA) which equally represents the regular grammar corresponding to the phosphorylation sites. Extensive experiments on several datasets generated by us reveal that, our inferred grammar successfully predicts phosphorylation sites in a kinase specific manner. It performs significantly better when compared with the other existing phosphorylation site prediction methods. We have also compared our inferred DSFA with two other GI inference algorithms. The DSFA generated by our method performs superior which indicates that our method is robust and has a potential for predicting the phosphorylation sites in a kinase specific manner.

Computer-based prediction of mitochondria-targeting peptides.

PubMed

Martelli, Pier Luigi; Savojardo, Castrense; Fariselli, Piero; Tasco, Gianluca; Casadio, Rita

2015-01-01

Computational methods are invaluable when protein sequences, directly derived from genomic data, need functional and structural annotation. Subcellular localization is a feature necessary for understanding the protein role and the compartment where the mature protein is active and very difficult to characterize experimentally. Mitochondrial proteins encoded on the cytosolic ribosomes carry specific patterns in the precursor sequence from where it is possible to recognize a peptide targeting the protein to its final destination. Here we discuss to which extent it is feasible to develop computational methods for detecting mitochondrial targeting peptides in the precursor sequences and benchmark our and other methods on the human mitochondrial proteins endowed with experimentally characterized targeting peptides. Furthermore, we illustrate our newly implemented web server and its usage on the whole human proteome in order to infer mitochondrial targeting peptides, their cleavage sites, and whether the targeting peptide regions contain or not arginine-rich recurrent motifs. By this, we add some other 2,800 human proteins to the 124 ones already experimentally annotated with a mitochondrial targeting peptide.

Environmental risk analysis of oil handling facilities in port areas. Application to Tarragona harbor (NE Spain).

PubMed

Valdor, Paloma F; Gómez, Aina G; Puente, Araceli

2015-01-15

Diffuse pollution from oil spills is a widespread problem in port areas (as a result of fuel supply, navigation and loading/unloading activities). This article presents a method to assess the environmental risk of oil handling facilities in port areas. The method is based on (i) identification of environmental hazards, (ii) characterization of meteorological and oceanographic conditions, (iii) characterization of environmental risk scenarios, and (iv) assessment of environmental risk. The procedure has been tested by application to the Tarragona harbor. The results show that the method is capable of representing (i) specific local pollution cases (i.e., discriminating between products and quantities released by a discharge source), (ii) oceanographic and meteorological conditions (selecting a representative subset data), and (iii) potentially affected areas in probabilistic terms. Accordingly, it can inform the design of monitoring plans to study and control the environmental impact of these facilities, as well as the design of contingency plans. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development and Validation of a Stability-Indicating Assay of Etofenamate by RP-HPLC and Characterization of Degradation Products

PubMed Central

Peraman, Ramalingam; Nayakanti, Devanna; Dugga, Hari Hara Theja; Kodikonda, Sudhakara

2013-01-01

A validated stability-indicating RP-HPLC method for etofenamate (ETF) was developed by separating its degradation products on a C18 (250 mm × 4.6 mm 5 μm) Qualisil BDS column using a phosphate buffer (pH-adjusted to 6.0 with orthophosphoric acid) and methanol in the ratio of 20:80 % v/v as the mobile phase at a flow rate of 1.0 mL/min. The column effluents were monitored by a photodiode array detector set at 286 nm. The method was validated in terms of specificity, linearity, accuracy, precision, detection limit, quantification limit, and robustness. Forced degradation of etofenamate was carried out under acidic, basic, thermal, photo, and peroxide conditions and the major degradation products of acidic and basic degradation were isolated and characterized by 1H-NMR, 13C-NMR, and mass spectral studies. The mass balance of the method varied between 92–99%. PMID:24482770

Bovine Papillomavirus in Brazil: Detection of Coinfection of Unusual Types by a PCR-RFLP Method

PubMed Central

Carvalho, R. F.; Sakata, S. T.; Giovanni, D. N. S.; Mori, E.; Brandão, P. E.; Richtzenhain, L. J.; Pozzi, C. R.; Arcaro, J. R. P.; Miranda, M. S.; Mazzuchelli-de-Souza, J.; Melo, T. C.; Comenale, G.; Assaf, S. L. M. R.; Beçak, W.; Stocco, R. C.

2013-01-01

Bovine papillomavirus (BPV) is recognized as a causal agent of benign and malignant tumors in cattle. Thirteen types of BPV are currently characterized and classified into three distinct genera, associated with different pathological outcomes. The described BPV types as well as other putative ones have been demonstrated by molecular biology methods, mainly by the employment of degenerated PCR primers. Specifically, divergences in the nucleotide sequence of the L1 gene are useful for the identification and classification of new papillomavirus types. On the present work, a method based on the PCR-RFLP technique and DNA sequencing was evaluated as a screening tool, allowing for the detection of two relatively rare types of BPV in lesions samples from a six-year-old Holstein dairy cow, chronically affected with cutaneous papillomatosis. These findings point to the dissemination of BPVs with unclear pathogenic potential, since two relatively rare, new described BPV types, which were first characterized in Japan, were also detected in Brazil. PMID:23865043

Rapid transcriptome characterization and parsing of sequences in a non-model host-pathogen interaction; pea-Sclerotinia sclerotiorum

PubMed Central

2012-01-01

Background White mold, caused by Sclerotinia sclerotiorum, is one of the most important diseases of pea (Pisum sativum L.), however, little is known about the genetics and biochemistry of this interaction. Identification of genes underlying resistance in the host or pathogenicity and virulence factors in the pathogen will increase our knowledge of the pea-S. sclerotiorum interaction and facilitate the introgression of new resistance genes into commercial pea varieties. Although the S. sclerotiorum genome sequence is available, no pea genome is available, due in part to its large genome size (~3500 Mb) and extensive repeated motifs. Here we present an EST data set specific to the interaction between S. sclerotiorum and pea, and a method to distinguish pathogen and host sequences without a species-specific reference genome. Results 10,158 contigs were obtained by de novo assembly of 128,720 high-quality reads generated by 454 pyrosequencing of the pea-S. sclerotiorum interactome. A method based on the tBLASTx program was modified to distinguish pea and S. sclerotiorum ESTs. To test this strategy, a mixture of known ESTs (18,490 pea and 17,198 S. sclerotiorum ESTs) from public databases were pooled and parsed; the tBLASTx method successfully separated 90.1% of the artificial EST mix with 99.9% accuracy. The tBLASTx method successfully parsed 89.4% of the 454-derived EST contigs, as validated by PCR, into pea (6,299 contigs) and S. sclerotiorum (2,780 contigs) categories. Two thousand eight hundred and forty pea ESTs and 996 S. sclerotiorum ESTs were predicted to be expressed specifically during the pea-S. sclerotiorum interaction as determined by homology search against 81,449 pea ESTs (from flowers, leaves, cotyledons, epi- and hypocotyl, and etiolated and light treated etiolated seedlings) and 57,751 S. sclerotiorum ESTs (from mycelia at neutral pH, developing apothecia and developing sclerotia). Among those ESTs specifically expressed, 277 (9.8%) pea ESTs were predicted to be involved in plant defense and response to biotic or abiotic stress, and 93 (9.3%) S. sclerotiorum ESTs were predicted to be involved in pathogenicity/virulence. Additionally, 142 S. sclerotiorum ESTs were identified as secretory/signal peptides of which only 21 were previously reported. Conclusions We present and characterize an EST resource specific to the pea-S. sclerotiorum interaction. Additionally, the tBLASTx method used to parse S. sclerotiorum and pea ESTs was demonstrated to be a reliable and accurate method to distinguish ESTs without a reference genome. PMID:23181755

Statistical power and utility of meta-analysis methods for cross-phenotype genome-wide association studies.

PubMed

Zhu, Zhaozhong; Anttila, Verneri; Smoller, Jordan W; Lee, Phil H

2018-01-01

Advances in recent genome wide association studies (GWAS) suggest that pleiotropic effects on human complex traits are widespread. A number of classic and recent meta-analysis methods have been used to identify genetic loci with pleiotropic effects, but the overall performance of these methods is not well understood. In this work, we use extensive simulations and case studies of GWAS datasets to investigate the power and type-I error rates of ten meta-analysis methods. We specifically focus on three conditions commonly encountered in the studies of multiple traits: (1) extensive heterogeneity of genetic effects; (2) characterization of trait-specific association; and (3) inflated correlation of GWAS due to overlapping samples. Although the statistical power is highly variable under distinct study conditions, we found the superior power of several methods under diverse heterogeneity. In particular, classic fixed-effects model showed surprisingly good performance when a variant is associated with more than a half of study traits. As the number of traits with null effects increases, ASSET performed the best along with competitive specificity and sensitivity. With opposite directional effects, CPASSOC featured the first-rate power. However, caution is advised when using CPASSOC for studying genetically correlated traits with overlapping samples. We conclude with a discussion of unresolved issues and directions for future research.

Emerging technologies for the non-invasive characterization of physical-mechanical properties of tablets.

PubMed

Dave, Vivek S; Shahin, Hend I; Youngren-Ortiz, Susanne R; Chougule, Mahavir B; Haware, Rahul V

2017-10-30

The density, porosity, breaking force, viscoelastic properties, and the presence or absence of any structural defects or irregularities are important physical-mechanical quality attributes of popular solid dosage forms like tablets. The irregularities associated with these attributes may influence the drug product functionality. Thus, an accurate and efficient characterization of these properties is critical for successful development and manufacturing of a robust tablets. These properties are mainly analyzed and monitored with traditional pharmacopeial and non-pharmacopeial methods. Such methods are associated with several challenges such as lack of spatial resolution, efficiency, or sample-sparing attributes. Recent advances in technology, design, instrumentation, and software have led to the emergence of newer techniques for non-invasive characterization of physical-mechanical properties of tablets. These techniques include near infrared spectroscopy, Raman spectroscopy, X-ray microtomography, nuclear magnetic resonance (NMR) imaging, terahertz pulsed imaging, laser-induced breakdown spectroscopy, and various acoustic- and thermal-based techniques. Such state-of-the-art techniques are currently applied at various stages of development and manufacturing of tablets at industrial scale. Each technique has specific advantages or challenges with respect to operational efficiency and cost, compared to traditional analytical methods. Currently, most of these techniques are used as secondary analytical tools to support the traditional methods in characterizing or monitoring tablet quality attributes. Therefore, further development in the instrumentation and software, and studies on the applications are necessary for their adoption in routine analysis and monitoring of tablet physical-mechanical properties. Copyright © 2017 Elsevier B.V. All rights reserved.

Usefulness and limitation of measurement methods for evaluation of tissue-engineered cartilage function and characterization using nanosecond pulsed laser

NASA Astrophysics Data System (ADS)

Ishihara, Miya; Sato, Masato; Kaneshiro, Nagatoshi; Mitani, Genya; Nagai, Toshihiro; Kutsuna, Toshiharu; Ishihara, Masayuki; Mochida, Joji; Kikuchi, Makoto

2007-02-01

There is a demand in the field of regenerative medicine for measurement technology that enables determination of functions and characterizations of engineered tissue. Regenerative medicine involving the articular cartilage in particular requires measurement of viscoelastic properties and characterization of the extracellular matrix, which plays a major role in articular cartilage. To meet this demand, we previously proposed a noninvasive method for determination of the viscoelasticity using laser-induced thermoelastic wave (1,2). We also proposed a method for characterization of the extracellular matrix using time-resolved autofluorescence spectroscopy, which could be performed simultaneously with laser-induced thermoelastic wave measurement(3). The purpose of this study was to verify the usefulness and limitation of these methods for evaluation of actual engineered cartilage. 3rd Q-SW Nd:YAG laser pulses, which are delivered through optical fiber, were used for the light source. Laser-induced thermoelastic waves were detected by a sensor consisting of a piezoelectric transducer, which was designed for use in arthroscopy(4). The time-resolved fluorescence spectroscopy was measured by a photonic multichannel analyzer with 4ch digital signal generator. Various tissue-engineered cartilages were developed as samples. Only a limited range of sample thickness could be measured, however, the measured viscoelastic parameters had a positive correlation with culture time, that is, the degree of formation of extracellular matrix(5,6). There were significant differences in the fluorescent parameters among the phenotypic expressions of cartilage because chondrocyte produces specific extracellular matrix as in collagen types depending on its phenotype.

Etching-free patterning method for electrical characterization of atomically thin MoSe2 films grown by chemical vapor deposition.

PubMed

Utama, M Iqbal Bakti; Lu, Xin; Zhan, Da; Ha, Son Tung; Yuan, Yanwen; Shen, Zexiang; Xiong, Qihua

2014-11-07

Patterning two-dimensional materials into specific spatial arrangements and geometries is essential for both fundamental studies of materials and practical applications in electronics. However, the currently available patterning methods generally require etching steps that rely on complicated and expensive procedures. We report here a facile patterning method for atomically thin MoSe2 films using stripping with an SU-8 negative resist layer exposed to electron beam lithography. Additional steps of chemical and physical etching were not necessary in this SU-8 patterning method. The SU-8 patterning was used to define a ribbon channel from a field effect transistor of MoSe2 film, which was grown by chemical vapor deposition. The narrowing of the conduction channel area with SU-8 patterning was crucial in suppressing the leakage current within the device, thereby allowing a more accurate interpretation of the electrical characterization results from the sample. An electrical transport study, enabled by the SU-8 patterning, showed a variable range hopping behavior at high temperatures.

Analysis of biomedical time signals for characterization of cutaneous diabetic micro-angiopathy

NASA Astrophysics Data System (ADS)

Kraitl, Jens; Ewald, Hartmut

2007-02-01

Photo-plethysmography (PPG) is frequently used in research on microcirculation of blood. It is a non-invasive procedure and takes minimal time to be carried out. Usually PPG time series are analyzed by conventional linear methods, mainly Fourier analysis. These methods may not be optimal for the investigation of nonlinear effects of the hearth circulation system like vasomotion, autoregulation, thermoregulation, breathing, heartbeat and vessels. The wavelet analysis of the PPG time series is a specific, sensitive nonlinear method for the in vivo identification of hearth circulation patterns and human health status. This nonlinear analysis of PPG signals provides additional information which cannot be detected using conventional approaches. The wavelet analysis has been used to study healthy subjects and to characterize the health status of patients with a functional cutaneous microangiopathy which was associated with diabetic neuropathy. The non-invasive in vivo method is based on the radiation of monochromatic light through an area of skin on the finger. A Photometrical Measurement Device (PMD) has been developed. The PMD is suitable for non-invasive continuous online monitoring of one or more biologic constituent values and blood circulation patterns.

Determination of Specific Losses in the Limbs of an Epstein Frame Using a Three Epstein Frame Methodology Applied to Grain Oriented Electrical Steels

PubMed Central

Parent, Guillaume; Penin, Rémi; Lecointe, Jean-Philippe; Brudny, Jean-François; Belgrand, Thierry

2016-01-01

An experimental method to characterize the magnetic properties of Grain Oriented Electrical Steel in the rolling direction is proposed in this paper. It relies on the use of three 25 cm Epstein frames combined to generate three test-frames of different lengths. This enables the identification of the effective specific losses of the electrical steel when magnetization is applied along the rolling direction. As a consequence, it evidences the deviation of the loss figures obtained using the standardised Epstein test. The difference in losses is explained by the fact that the described method gives “only” the losses attached to the straight parts. The concept of the magnetic path length as defined by the standard is discussed. PMID:27271637

Studies of proteoglycan involvement in CPP-mediated delivery.

PubMed

Wittrup, Anders; Zhang, Si-He; Belting, Mattias

2011-01-01

Cell-penetrating peptides (CPPs) are widely used to deliver macromolecular cargoes to intracellular sites of action. Many CPPs have been demonstrated to rely on cell surface heparan sulfate proteoglycans (HSPGs) for efficient cellular entry and delivery. In this chapter, we describe methods for the study of PG involvement in CPP uptake. We provide descriptions of how to determine whether uptake of a CPP of interest is dependent on PGs. We also provide detailed protocols for the purification of PGs by anion-exchange chromatography as well as the characterization of the HSPG core protein composition of a cell line of interest. Finally, we present methods for modulating the expression level of specific HSPG core proteins as a means to determine the core protein specificity in the uptake of a particular CPP.

Determination of Specific Losses in the Limbs of an Epstein Frame Using a Three Epstein Frame Methodology Applied to Grain Oriented Electrical Steels.

PubMed

Parent, Guillaume; Penin, Rémi; Lecointe, Jean-Philippe; Brudny, Jean-François; Belgrand, Thierry

2016-06-04

An experimental method to characterize the magnetic properties of Grain Oriented Electrical Steel in the rolling direction is proposed in this paper. It relies on the use of three 25 cm Epstein frames combined to generate three test-frames of different lengths. This enables the identification of the effective specific losses of the electrical steel when magnetization is applied along the rolling direction. As a consequence, it evidences the deviation of the loss figures obtained using the standardised Epstein test. The difference in losses is explained by the fact that the described method gives "only" the losses attached to the straight parts. The concept of the magnetic path length as defined by the standard is discussed.

Detection of anti-neutrophil antibodies in autoimmune neutropenia of infancy: a multicenter study.

PubMed

Sella, Ruti; Flomenblit, Lena; Goldstein, Itamar; Kaplinsky, Chaim

2010-02-01

Autoimmune neutropenia of infancy is caused by neutrophil-specific autoantibodies. Primary AIN is characterized by neutrophil count < 500 ml and a benign self-limiting course. Detecting specific antibodies against the polymorphic human neutrophil antigen usually confirms the diagnosis. Current available tests, however, are expensive and inapplicable in many laboratories as they require the use of isolated and fixed granulocytes obtained from donors pretyped for their distinct HNA alloform. To assess the performance of a modified test to identify by FACS-analysis granulocyte-specific antibodies in the sera of neutropenic children. We evaluated 120 children with a clinical suspicion of AIN, whose sera were analyzed by flow cytometry for the presence of autoantibodies using the indirect granulocyte immunofluorescence test. In contrast to the traditional tests, the sera were tested against randomly selected untyped neutrophils derived from a batch of 10 anonymous healthy subjects, presumably including the common HNA alloforms. Control sera samples were from patients with chemotherapy-induced, familial or congenital neutropenias. To further assure the quality of the new test, we retested six samples previously tested by the gold standard method. All medical files were screened and clinical outcomes were recorded. Our method showed specificity of 85%, sensitivity of 62.5%, and a positive predictive value of 91.8%, values quite similar to those obtained by more traditional methods. The new method showed high specificity for detection of anti-neutrophil antibodies in the appropriate clinical setting and could be an effective tool for clinical decision making.

Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides.

PubMed

Mishima, Eikan; Jinno, Daisuke; Akiyama, Yasutoshi; Itoh, Kunihiko; Nankumo, Shinnosuke; Shima, Hisato; Kikuchi, Koichi; Takeuchi, Yoichi; Elkordy, Alaa; Suzuki, Takehiro; Niizuma, Kuniyasu; Ito, Sadayoshi; Tomioka, Yoshihisa; Abe, Takaaki

2015-01-01

The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism.

U.S. Geological Survey research in radioactive waste disposal - Fiscal years 1986-1990

USGS Publications Warehouse

Trask, N.J.; Stevens, P.R.

1991-01-01

The report summarizes progress on geologic and hydrologic research related to the disposal of radioactive wastes. The research efforts are categorized according to whether they are related most directly to: (1) high-level wastes, (2) transuranic wastes, (3) low-level and mixed low-level and hazardous wastes, or (4) uranium mill tailings. Included is research applicable to the identification and geohydrologic characterization of waste-disposal sites, to investigations of specific sites where wastes have been stored, to development of techniques and methods for characterizing disposal sites, and to studies of geologic and hydrologic processes related to the transport and/or retention of waste radionuclides.

Purification of Plant Receptor Kinases from Plant Plasma Membranes.

PubMed

Lee, Jin Suk

2017-01-01

Receptor kinases play a central role in various biological processes, but due to their low abundance and highly hydrophobic and dynamic nature, only a few of them have been functionally characterized, and their partners and ligands remain unidentified. Receptor protein extraction and purification from plant tissues is one of the most challenging steps for the success of various biochemical analyses to characterize their function. Immunoprecipitation is a widely used and selective method for enriching or purifying a specific protein. Here we describe two different optimized protein purification protocols, batch and on-chip immunoprecipitation, which efficiently isolate plant membrane receptor kinases for functional analysis.

Environmental exposures and health impacts of PFAS ...

EPA Pesticide Factsheets

Environmental exposures and health impacts of PFAS The National Exposure Research Laboratory (NERL) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA mission to protect human health and the environment. HEASD research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.

Preparation of porous Si and TiO 2 nanofibres using a sulphur-templating method for lithium storage

DOE PAGES

McCormac, Kathleen; Byrd, Ian; Brannen, Rodney; ...

2015-02-03

We prepared highly porous Si/TiO 2 composite nanofibres using a unique sulphur-templating method combined with electrospinning. The structure, morphology, surface area, phase and composition of these nanofibres were characterized using Raman spectroscopy, scanning electron microscopy, powder X-ray diffraction, surface area analyser and thermogravimetric analyser. The specific surface area of Si/TiO 2 porous NFs is as large as 387m 2g -1, whose silicon capacity can be maintained above 1580mAhg -1 in 180 cycles.

UV-Visible and Infrared Methods for Investigating Lipid-Rhodopsin Membrane Interactions

PubMed Central

Brown, Michael F.

2017-01-01

Summary Experimental UV-visible and Fourier transform infrared (FTIR) spectroscopic methods are described for characterizing lipid-protein interactions for the example of rhodopsin in a membrane bilayer environment. The combined use of FTIR and UV-visible difference spectroscopy monitors the structural and functional changes during rhodopsin activation. Such studies investigate how membrane lipids stabilize the various rhodopsin photoproducts, analogous to mutating the protein. Interpretation of the results entails a non-specific flexible surface model for explaining the role of membrane lipid-protein interactions in biological functions. PMID:22976026

Electrical network method for the thermal or structural characterization of a conducting material sample or structure

DOEpatents

Ortiz, Marco G.

1993-01-01

A method for modeling a conducting material sample or structure system, as an electrical network of resistances in which each resistance of the network is representative of a specific physical region of the system. The method encompasses measuring a resistance between two external leads and using this measurement in a series of equations describing the network to solve for the network resistances for a specified region and temperature. A calibration system is then developed using the calculated resistances at specified temperatures. This allows for the translation of the calculated resistances to a region temperature. The method can also be used to detect and quantify structural defects in the system.

Electrical network method for the thermal or structural characterization of a conducting material sample or structure

DOEpatents

Ortiz, M.G.

1993-06-08

A method for modeling a conducting material sample or structure system, as an electrical network of resistances in which each resistance of the network is representative of a specific physical region of the system. The method encompasses measuring a resistance between two external leads and using this measurement in a series of equations describing the network to solve for the network resistances for a specified region and temperature. A calibration system is then developed using the calculated resistances at specified temperatures. This allows for the translation of the calculated resistances to a region temperature. The method can also be used to detect and quantify structural defects in the system.

Characterization of Biofilm Community Structure by Ribosomal RNA sequences

DTIC Science & Technology

1989-12-01

for strains of Fibrobacter, 2) Desulfobacter genus-specific probe, 3) Desulfosarcina genus-specific probe, 4) archaebacterial kingdom -specific probes...and 5) eubacterial kingdom -specific probes 5) eukaryote kingdom -specific probe and 6) a general probe encompassing all characterized sulfate-reducing...sets have been fabricated. The group-specific primer sets selectively amplify either sulfate-reducing bacteria or archaebacteria . The SRB-specific

Photogrammetry and Videogrammetry Methods for Solar Sails and Other Gossamer Structures

NASA Technical Reports Server (NTRS)

Black, Jonathan T.; Pappa, Richard S.

2004-01-01

Ultra-lightweight and inflatable gossamer space structures are designed to be tightly packaged for launch, then deploy or inflate once in space. These properties will allow for in-space construction of very large structures 10 to 1000 meters in size such as solar sails, inflatable antennae, and space solar power stations using a single launch. Solar sails are of particular interest because of their potential for propellantless propulsion. Gossamer structures do, however, have significant complications. Their low mass and high flexibility make them very difficult to test on the ground. The added mass and stiffness of attached measurement devices can significantly alter the static and dynamic properties of the structure. This complication necessitates an alternative approach for characterization. This paper discusses the development and application of photogrammetry and videogrammetry methods for the static and dynamic characterization of gossamer structures, as four specific solar sail applications demonstrate. The applications prove that high-resolution, full-field, non-contact static measurements of solar sails using dot projection photogrammetry are possible as well as full-field, noncontact, dynamic characterization using dot projection videogrammetry.

Extraction and Characterization of Surfactants from Atmospheric Aerosols.

PubMed

Nozière, Barbara; Gérard, Violaine; Baduel, Christine; Ferronato, Corinne

2017-04-21

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in the Earth's atmosphere, a central process in meteorology, hydrology, and for the climate system. But because specific extraction and characterization of these compounds have been lacking for decades, very little is known on their identity, properties, mode of action and origins, thus preventing the full understanding of cloud formation and its potential links with the Earth's ecosystems. In this paper we present recently developed methods for 1) the targeted extraction of all the surfactants from atmospheric aerosol samples and for the determination of 2) their absolute concentrations in the aerosol phase and 3) their static surface tension curves in water, including their Critical Micelle Concentration (CMC). These methods have been validated with 9 references surfactants, including anionic, cationic and non-ionic ones. Examples of results are presented for surfactants found in fine aerosol particles (diameter <1 μm) collected at a coastal site in Croatia and suggestions for future improvements and other characterizations than those presented are discussed.

Characterization of Haemophilus parasuis isolated from Brazilian swine through serotyping, AFLP and PFGE.

PubMed

Castilla, Karina Salvagni; de Gobbi, Débora Dirani Sena; Moreno, Luisa Zanolli; Paixão, Renata; Coutinho, Tania Alen; dos Santos, José Lúcio; Moreno, Andrea Micke

2012-06-01

Haemophilus parasuis infection in pigs is characterized by fibrinous polyserositis, arthritis and meningitis. Despite the fact that traditional diagnosis is based on herd history, clinical signs, bacterial isolation and serotyping, molecular-based methods are alternatives for species-specific tests and epidemiological studies. The aim of this study was to characterize H. parasuis field strains from different states of Brazil, employing serotyping and genotyping methods. Serotyping revealed that serovar 4 was the most prevalent (26.1%), followed by serovars 5 (17.4%), 14 (8.7%), 13 (4.4%) and 2 (4.4%), whereas 39% of the strains were considered as untypeable. AFLP with a single enzyme and PFGE were able to type all isolates tested, generating 34 and 20 different profiles, respectively, including untypeable strains. Besides the slightly higher discrimination index presented by AFLP, PFGE with Not I restriction enzyme showed a better correlation with epidemiological data, grouping strains of the same serovar, animal or farm origin. The results indicated AFLP and PFGE as valuable tools for typing H. parasuis isolates collected in Brazil. Copyright © 2011 Elsevier Ltd. All rights reserved.

A novel image-based quantitative method for the characterization of NETosis

PubMed Central

Zhao, Wenpu; Fogg, Darin K.; Kaplan, Mariana J.

2015-01-01

NETosis is a newly recognized mechanism of programmed neutrophil death. It is characterized by a stepwise progression of chromatin decondensation, membrane rupture, and release of bactericidal DNA-based structures called neutrophil extracellular traps (NETs). Conventional ‘suicidal’ NETosis has been described in pathogenic models of systemic autoimmune disorders. Recent in vivo studies suggest that a process of ‘vital’ NETosis also exists, in which chromatin is condensed and membrane integrity is preserved. Techniques to assess ‘suicidal’ or ‘vital’ NET formation in a specific, quantitative, rapid and semiautomated way have been lacking, hindering the characterization of this process. Here we have developed a new method to simultaneously assess both ‘suicidal’ and ‘vital’ NETosis, using high-speed multi-spectral imaging coupled to morphometric image analysis, to quantify spontaneous NET formation observed ex-vivo or stimulus-induced NET formation triggered in vitro. Use of imaging flow cytometry allows automated, quantitative and rapid analysis of subcellular morphology and texture, and introduces the potential for further investigation using NETosis as a biomarker in pre-clinical and clinical studies. PMID:26003624

Extraction and Characterization of Surfactants from Atmospheric Aerosols

PubMed Central

Baduel, Christine; Ferronato, Corinne

2017-01-01

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in the Earth's atmosphere, a central process in meteorology, hydrology, and for the climate system. But because specific extraction and characterization of these compounds have been lacking for decades, very little is known on their identity, properties, mode of action and origins, thus preventing the full understanding of cloud formation and its potential links with the Earth's ecosystems. In this paper we present recently developed methods for 1) the targeted extraction of all the surfactants from atmospheric aerosol samples and for the determination of 2) their absolute concentrations in the aerosol phase and 3) their static surface tension curves in water, including their Critical Micelle Concentration (CMC). These methods have been validated with 9 references surfactants, including anionic, cationic and non-ionic ones. Examples of results are presented for surfactants found in fine aerosol particles (diameter <1 μm) collected at a coastal site in Croatia and suggestions for future improvements and other characterizations than those presented are discussed. PMID:28518073

Purification and high-resolution top-down mass spectrometric characterization of human salivary α-amylase.

PubMed

Peng, Ying; Chen, Xin; Sato, Takuya; Rankin, Scott A; Tsuji, Ryohei F; Ge, Ying

2012-04-03

Human salivary α-amylase (HSAMY) is a major component of salivary secretions, possessing multiple important biological functions. Here we have established three methods to purify HSAMY in human saliva for comprehensive characterization of HSAMY by high-resolution top-down mass spectrometry (MS). Among the three purification methods, the affinity method based on the enzyme-substrate specific interaction between amylase and glycogen is preferred, providing the highest purity HSAMY with high reproducibility. Subsequently, we employed Fourier transform ion cyclotron resonance MS to analyze the purified HSAMY. The predominant form of α-amylase purified from saliva of various races and genders is nonglycosylated with the same molecular weight of 55,881.2, which is 1885.8 lower than the calculated value based on the DNA-predicted sequence. High-resolution MS revealed the truncation of the first 15 N-terminal amino acids (-1858.96) and the subsequent formation of pyroglutamic acid at the new N-terminus Gln (-17.03). More importantly, five disulfide bonds in HSAMY were identified (-10.08) and effectively localized by tandem MS in conjunction with complete and partial reduction by tris (2-carboxyethyl) phosphine. Overall, this study demonstrates that top-down MS combined with affinity purification and partial reduction is a powerful method for rapid purification and complete characterization of large proteins with complex and overlapping disulfide bond patterns.

Comparative Study in Laboratory Rats to Validate Sperm Quality Methods and Endpoints

NASA Technical Reports Server (NTRS)

Price, W. A.; Briggs, G. B.; Alexander, W. K.; Still, K. R.; Grasman, K. A.

2000-01-01

Abstract The Naval Health Research Center, Detachment (Toxicology) performs toxicity studies in laboratory animals to characterize the risk of exposure to chemicals of Navy interest. Research was conducted at the Toxicology Detachment at WPAFB, OH in collaboration with Wright State University, Department of Biological Sciences for the validation of new bioassay methods for evaluating reproductive toxicity. The Hamilton Thorne sperm analyzer was used to evaluate sperm damage produced by exposure to a known testicular toxic agent, methoxyacetic acid and by inhalation exposure to JP-8 and JP-5 in laboratory rats. Sperm quality parameters were evaluated (sperm concentration, motility, and morphology) to provide evidence of sperm damage. The Hamilton Thorne sperm analyzer utilizes a DNA specific fluorescent stain (similar to flow cytometry) and digitized optical computer analysis to detect sperm cell damage. The computer assisted sperm analysis (CASA) is a more rapid, robust, predictive and sensitive method for characterizing reproductive toxicity. The results presented in this poster report validation information showing exposure to methoxyacetic acid causes reproductive toxicity and inhalation exposure to JP-8 and JP-5 had no significant effects. The CASA method detects early changes that result in reproductive deficits and these data will be used in a continuing program to characterize the toxicity of chemicals, and combinations of chemicals, of military interest to formulate permissible exposure limits.

Synthesis and characterization of the Cu2ZnSnS4 system for photovoltaic applications

NASA Astrophysics Data System (ADS)

Sánchez Pinzón, D. L.; Soracá Perez, G. Y.; Gómez Cuaspud, J. A.; López, E. Vera

2017-01-01

This paper focuses on the synthesis and characterization of a ceramic material based on the Cu2ZnSnS4 system, through the implementation of a hydrothermal route. For this purpose, we started from nitrate dissolutions in a 1.0mol L-1 concentration, which were mixed and treated in a teflon lined vessel steel at 280°C for 48h. The Physicochemical characterization of the solid was evaluated by means of ultraviolet visible spectroscopy (UV-VIS), X-ray diffraction (XRD), Raman spectroscopy, scanning and transmission electron microscopy (SEM-TEM) and solid state impedance spectroscopy (IS). The initial characterization through UV measurements confirms a Band-gap around 1.46eV obtained by the Kubelka-Munk method, which demonstrates the effectiveness of the synthesis method in the obtaining of a semiconductor material. The XRD results confirm the obtaining of a crystalline material of pure phase with tetragonal geometry and I-42m space group. The preferential crystalline orientation was achieved along (2 2 0) facet, with crystallite sizes of nanometric order (6.0nm). The morphological aspects evaluated by means electron microscopy, confirmed the homogeneity of the material, showing specifically a series of textural and surface properties of relevant importance. Finally, the electrical characterizations allow to validate the semiconductor behaviour of CZTS system for development of photovoltaic technologies.

Structure of adenovirus bound to cellular receptor car

DOEpatents

Freimuth, Paul I.

2007-01-02

Disclosed is a mutant CAR-DI-binding adenovirus which has a genome comprising one or more mutations in sequences which encode the fiber protein knob domain wherein the mutation causes the encoded viral particle to have a significantly weakened binding affinity for CAR-DI relative to wild-type adenovirus. Such mutations may be in sequences which encode either the AB loop, or the HI loop of the fiber protein knob domain. Specific residues and mutations are described. Also disclosed is a method for generating a mutant adenovirus which is characterized by a receptor binding affinity or specificity which differs substantially from wild type.

Coupling heat and chemical tracer experiments for estimating heat transfer parameters in shallow alluvial aquifers.

PubMed

Wildemeersch, S; Jamin, P; Orban, P; Hermans, T; Klepikova, M; Nguyen, F; Brouyère, S; Dassargues, A

2014-11-15

Geothermal energy systems, closed or open, are increasingly considered for heating and/or cooling buildings. The efficiency of such systems depends on the thermal properties of the subsurface. Therefore, feasibility and impact studies performed prior to their installation should include a field characterization of thermal properties and a heat transfer model using parameter values measured in situ. However, there is a lack of in situ experiments and methodology for performing such a field characterization, especially for open systems. This study presents an in situ experiment designed for estimating heat transfer parameters in shallow alluvial aquifers with focus on the specific heat capacity. This experiment consists in simultaneously injecting hot water and a chemical tracer into the aquifer and monitoring the evolution of groundwater temperature and concentration in the recovery well (and possibly in other piezometers located down gradient). Temperature and concentrations are then used for estimating the specific heat capacity. The first method for estimating this parameter is based on a modeling in series of the chemical tracer and temperature breakthrough curves at the recovery well. The second method is based on an energy balance. The values of specific heat capacity estimated for both methods (2.30 and 2.54MJ/m(3)/K) for the experimental site in the alluvial aquifer of the Meuse River (Belgium) are almost identical and consistent with values found in the literature. Temperature breakthrough curves in other piezometers are not required for estimating the specific heat capacity. However, they highlight that heat transfer in the alluvial aquifer of the Meuse River is complex and contrasted with different dominant process depending on the depth leading to significant vertical heat exchange between upper and lower part of the aquifer. Furthermore, these temperature breakthrough curves could be included in the calibration of a complex heat transfer model for estimating the entire set of heat transfer parameters and their spatial distribution by inverse modeling. Copyright © 2014 Elsevier B.V. All rights reserved.

Solid State NMR Characterization of Ibuprofen:Nicotinamide Cocrystals and New Idea for Controlling Release of Drugs Embedded into Mesoporous Silica Particles.

PubMed

Skorupska, Ewa; Kaźmierski, Sławomir; Potrzebowski, Marek J

2017-05-01

Grinding and melting methods were employed for synthesis of pharmaceutical cocrystals formed by racemic (R/S) and entiomeric (S) ibuprofen (IBU) and nicotinamide (NA) as coformer. Obtained (R/S)-IBU:NA and (S)-IBU:NA cocrystals were fully characterized by means of advanced one- and two-dimensional solid state nuclear magnetic resonance (SS NMR) techniques with very fast magic angle spinning (MAS) at 60 kHz. The distinction in molecular packing and specific hydrogen bonding pattern was clearly recognized by analysis of 1 H, 13 C, and 15 N spectra. It is concluded from these studies that both methods (grinding and melting) provide exactly the same, specific forms of cocrystals. Thermal solvent-free (TSF) approach was used for loading of (R/S)-IBU:NA and (S)-IBU:NA into the pores of MCM-41 mesoporous silica particle (MSP). The progress and efficiency of this process was analyzed by NMR spectroscopy. It has been confirmed that TSF method is an effective and safe technique of filling the MSP pores with active pharmaceutical ingredients (APIs). By analyzing the NMR results, it has been further proved that excess of IBU and NA components, which are not embedded into the pores during melting and cooling, crystallize on the MCM-41 walls preserving very specific arrangement, characteristic for crystalline samples. By investigating kinetic of release for (R/S)-IBU/MCM-41, (S)-IBU:NA/MCM-41, and (R/S)-IBU:NA/MCM-41 samples containing active components exclusively inside of the pores, it was revealed that release of IBU is much faster for the first of the samples compared to those containing IBU and NA inside the pores. The hypothesis that the rate of release of API can be controlled by specific composition of cocrystal embedded into the MSP pore was further supported by study of (R/S)-IBU:BA/MCM-41 sample with benzoic acid (BA) as coformer.

Rapid polymerase chain reaction screening of Helicobacter pylori chromosomal point mutations.

PubMed

Ge, Z; Taylor, D E

1997-09-01

Microdiversity (within individual genes) in the genomes of different Helicobacter pylori strains has been demonstrated to be more frequent than that seen in other prokaryotes. Point mutations in some genes, such as the vacA and 23S ribosomal RNA genes could result in the alteration of pathogenicity or antibiotic susceptibility of individual H. pylori strains. Development of a simple, rapid, and reliable screening method would be useful in the molecular characterization of genetic variation among different H. pylori strains. The copP gene from H. pylori UA802 was used as a model for developing a mutation screening method. Four point mutations were introduced into the copP gene by in vitro site-directed mutagenesis and were verified by DNA sequencing. The mutated copP gene replaced the wild-type locus by natural transformation and homologous recombination. The site-specific mutants were screened by polymerase chain reaction (PCR) using 3'-end mismatched primers. The origins of the PCR fragments were demonstrated by Southern hybridization with the copP-derived DNA probe. Three of these four mutations were characterized by PCR with the specific primers that contained the 3'-terminal nucleotide complementary only to the mutated nucleotide on both plasmid and chromosomal DNA templates. One mutation was able to be identified with the foregoing primer containing an additional wild-type nucleotide at its 3'-end. Point mutant screening with these specific primers offers 100% sensitivity in the aforementioned conditions. To achieve optimal screening, the concentration of magnesium and the annealing temperature have to be adjusted. The procedure reported in this study is a simple, economical, rapid, and efficient approach in the identification of site-specific mutations on both plasmids and chromosomal DNA. Although the method was developed by using a specified H. pylori gene, it can be extended easily to other genes of interest in H. pylori or other organisms.

Use of PCR with Sequence-specific Primers for High-Resolution Human Leukocyte Antigen Typing of Patients with Narcolepsy

PubMed Central

Woo, Hye In; Joo, Eun Yeon; Lee, Kyung Wha

2012-01-01

Background Narcolepsy is a neurologic disorder characterized by excessive daytime sleepiness, symptoms of abnormal rapid eye movement (REM) sleep, and a strong association with HLA-DRB1*1501, -DQA1*0102, and -DQB1*0602. Here, we investigated the clinico-physical characteristics of Korean patients with narcolepsy, their HLA types, and the clinical utility of high-resolution PCR with sequence-specific primers (PCR-SSP) as a simple typing method for identifying DRB1*15/16, DQA1, and DQB1 alleles. Methods The study population consisted of 67 consecutively enrolled patients having unexplained daytime sleepiness and diagnosed narcolepsy based on clinical and neurological findings. Clinical data and the results of the multiple sleep latency test and polysomnography were reviewed, and HLA typing was performed using both high-resolution PCR-SSP and sequence-based typing (SBT). Results The 44 narcolepsy patients with cataplexy displayed significantly higher frequencies of DRB1*1501 (Pc= 0.003), DQA1*0102 (Pc=0.001), and DQB1*0602 (Pc=0.014) than the patients without cataplexy. Among patients carrying DRB1*1501-DQB1*0602 or DQA1*0102, the frequencies of a mean REM sleep latency of less than 20 min in nocturnal polysomnography and clinical findings, including sleep paralysis and hypnagogic hallucination were significantly higher. SBT and PCR-SSP showed 100% concordance for high-resolution typing of DRB1*15/16 alleles and DQA1 and DQB1 loci. Conclusions The clinical characteristics and somnographic findings of narcolepsy patients were associated with specific HLA alleles, including DRB1*1501, DQA1*0102, and DQB1*0602. Application of high-resolution PCR-SSP, a reliable and simple method, for both allele- and locus-specific HLA typing of DRB1*15/16, DQA1, and DQB1 would be useful for characterizing clinical status among subjects with narcolepsy. PMID:22259780

Windows to the soul: vision science as a tool for studying biological mechanisms of information processing deficits in schizophrenia.

PubMed

Yoon, Jong H; Sheremata, Summer L; Rokem, Ariel; Silver, Michael A

2013-10-31

Cognitive and information processing deficits are core features and important sources of disability in schizophrenia. Our understanding of the neural substrates of these deficits remains incomplete, in large part because the complexity of impairments in schizophrenia makes the identification of specific deficits very challenging. Vision science presents unique opportunities in this regard: many years of basic research have led to detailed characterization of relationships between structure and function in the early visual system and have produced sophisticated methods to quantify visual perception and characterize its neural substrates. We present a selective review of research that illustrates the opportunities for discovery provided by visual studies in schizophrenia. We highlight work that has been particularly effective in applying vision science methods to identify specific neural abnormalities underlying information processing deficits in schizophrenia. In addition, we describe studies that have utilized psychophysical experimental designs that mitigate generalized deficit confounds, thereby revealing specific visual impairments in schizophrenia. These studies contribute to accumulating evidence that early visual cortex is a useful experimental system for the study of local cortical circuit abnormalities in schizophrenia. The high degree of similarity across neocortical areas of neuronal subtypes and their patterns of connectivity suggests that insights obtained from the study of early visual cortex may be applicable to other brain regions. We conclude with a discussion of future studies that combine vision science and neuroimaging methods. These studies have the potential to address pressing questions in schizophrenia, including the dissociation of local circuit deficits vs. impairments in feedback modulation by cognitive processes such as spatial attention and working memory, and the relative contributions of glutamatergic and GABAergic deficits.

Scaling of the flow-stiffness relationship in weakly correlated single fractures

NASA Astrophysics Data System (ADS)

Petrovitch, Christopher L.

The remote characterization of the hydraulic properties of fractures in rocks is important in many subsurface projects. Fractures create uncertainty in the hydraulic properties of the subsurface in that their topology controls the amount of flow that can occur in addition to that from the matrix. In turn, the fracture topology is also affected by stress which alters the topology as the stress changes directly. This alteration of fracture topology with stress is captured by fracture specific stiffness. The specific stiffness of a single fracture can be remotely probed from the attenuation and velocity of seismic waves. The hydromechanical coupling of single fractures, i.e. the relationship between flow and stiffness, holds the key to finding a method to remotely characterize a fractures hydraulic properties. This thesis is separated into two parts: (1) a description of the hydromechanical coupling of fractures based on numerical models used to generate synthetic fractures, compute the flow through a fracture, and deform fracture topologies to unravel the scaling function that is fundamental to the hydromechanical coupling of single fractures; (2) a Discontinuous Galerkin (DG) method was developed to accurately simulate the scattered seismic waves from realistic fracture topologies. The scaling regimes of fluid flow and specific stiffness in weakly correlated fractures are identified by using techniques from Percolation Theory and initially treating the two processes separately. The fixed points associated with fluid flow were found to display critical scaling while the fixed points for specific stiffness were trivial. The two processes could be indirectly related because the trivial scaling of the mechanical properties allowed the specific stiffness to be used as surrogate to the void area fraction. The dynamic transport exponent was extracted at threshold by deforming fracture geometries within the effective medium regime (near the ``cubic law'' regime) to the critical regime. From this, a scaling function was defined for the hydromechanical coupling. This scaling function provides the link between fluid flow and fracture specific stiffness so that seismic waves may be used to remotely probe the hydraulic properties of fractures. Then, the DG method is shown to be capable of measuring such fracture specific stiffnesses by numerically measuring the velocity of interface waves when propagated across laboratory measured fracture geometries of Austin Chalk.

Detection of blob objects in microscopic zebrafish images based on gradient vector diffusion.

PubMed

Li, Gang; Liu, Tianming; Nie, Jingxin; Guo, Lei; Malicki, Jarema; Mara, Andrew; Holley, Scott A; Xia, Weiming; Wong, Stephen T C

2007-10-01

The zebrafish has become an important vertebrate animal model for the study of developmental biology, functional genomics, and disease mechanisms. It is also being used for drug discovery. Computerized detection of blob objects has been one of the important tasks in quantitative phenotyping of zebrafish. We present a new automated method that is able to detect blob objects, such as nuclei or cells in microscopic zebrafish images. This method is composed of three key steps. The first step is to produce a diffused gradient vector field by a physical elastic deformable model. In the second step, the flux image is computed on the diffused gradient vector field. The third step performs thresholding and nonmaximum suppression based on the flux image. We report the validation and experimental results of this method using zebrafish image datasets from three independent research labs. Both sensitivity and specificity of this method are over 90%. This method is able to differentiate closely juxtaposed or connected blob objects, with high sensitivity and specificity in different situations. It is characterized by a good, consistent performance in blob object detection.

Mean-Reverting Portfolio With Budget Constraint

NASA Astrophysics Data System (ADS)

Zhao, Ziping; Palomar, Daniel P.

2018-05-01

This paper considers the mean-reverting portfolio design problem arising from statistical arbitrage in the financial markets. We first propose a general problem formulation aimed at finding a portfolio of underlying component assets by optimizing a mean-reversion criterion characterizing the mean-reversion strength, taking into consideration the variance of the portfolio and an investment budget constraint. Then several specific problems are considered based on the general formulation, and efficient algorithms are proposed. Numerical results on both synthetic and market data show that our proposed mean-reverting portfolio design methods can generate consistent profits and outperform the traditional design methods and the benchmark methods in the literature.

Applications of Evolutionary Algorithms to Electromagnetic Materials Characterization and Design Problems

NASA Astrophysics Data System (ADS)

Frasch, Jonathan Lemoine

Determining the electrical permittivity and magnetic permeability of materials is an important task in electromagnetics research. The method using reflection and transmission scattering parameters to determine these constants has been widely employed for many years, ever since the work of Nicolson, Ross, and Weir in the 1970's. For general materials that are homogeneous, linear, and isotropic, the method they developed (the NRW method) works very well and provides an analytical solution. For materials which possess a metal backing or are applied as a coating to a metal surface, it can be difficult or even impossible to obtain a transmission measurement, especially when the coating is thin. In such a circumstance, it is common to resort to a method which uses two reflection type measurements. There are several such methods for free-space measurements, using multiple angles or polarizations for example. For waveguide measurements, obtaining two independent sources of information from which to extract two complex parameters can be a challenge. This dissertation covers three different topics. Two of these involve different techniques to characterize conductor-backed materials, and the third proposes a method for designing synthetic validation standards for use with standard NRW measurements. All three of these topics utilize modal expansions of electric and magnetic fields to analyze propagation in stepped rectangular waveguides. Two of the projects utilize evolutionary algorithms (EA) to design waveguide structures. These algorithms were developed specifically for these projects and utilize fairly recent innovations within the optimization community. The first characterization technique uses two different versions of a single vertical step in the waveguide. Samples to be tested lie inside the steps with the conductor reflection plane behind them. If the two reflection measurements are truly independent it should be possible to recover the values of two complex parameters, but success of the technique ultimately depends upon how independent the measurements actually are. Next, a method is demonstrated for developing synthetic verification standards. These standards are created from combinations of vertical steps formed from a single piece of metal or metal coated plastic. These fully insertable structures mimic some of the measurement characteristics of typical lab specimens and thus provide a useful tool for verifying the proper calibration and function of the experimental setup used for NRW characterization. These standards are designed with the use an EA, which compares possible designs based on the quality of the match with target parameter values. Several examples have been fabricated and tested, and the design specifications and results are presented. Finally, a second characterization technique is considered. This method uses multiple vertical steps to construct an error reducing structure within the waveguide, which allows parameters to be reliably extracted using both reflection and transmission measurements. These structures are designed with an EA, measuring fitness by the reduction of error in the extracted parameters. An additional EA is used to assist in the extraction of the material parameters supplying better initial guesses to a secant method solver. This hybrid approach greatly increases the stability of the solver and increases the speed of parameter extractions. Several designs have been identified and are analyzed.

Development of a Sequence-Characterized Amplified Region Marker-Targeted Quantitative PCR Assay for Strain-Specific Detection of Oenococcus oeni during Wine Malolactic Fermentation▿

PubMed Central

Solieri, Lisa; Giudici, Paolo

2010-01-01

Control over malolactic fermentation (MLF) is a difficult goal in winemaking and needs rapid methods to monitor Oenococcus oeni malolactic starters (MLS) in a stressful environment such as wine. In this study, we describe a novel quantitative PCR (QPCR) assay enabling the detection of an O. oeni strain during MLF without culturing. O. oeni strain LB221 was used as a model to develop a strain-specific sequence-characterized amplified region (SCAR) marker derived from a discriminatory OPA20-based randomly amplified polymorphic DNA (RAPD) band. The 5′ and 3′ flanking regions and the copy number of the SCAR marker were characterized using inverse PCR and Southern blotting, respectively. Primer pairs targeting the SCAR sequence enabled strain-specific detection without cross amplification of other O. oeni strains or wine species of lactic acid bacteria (LAB), acetic acid bacteria (AAB), and yeasts. The SCAR-QPCR assay was linear over a range of cell concentrations (7 log units) and detected as few as 2.2 × 102 CFU per ml of red wine with good quantification effectiveness, as shown by the correlation of QPCR and plate counting results. Therefore, the cultivation-independent monitoring of a single O. oeni strain in wine based on a SCAR marker represents a rapid and effective strain-specific approach. This strategy can be adopted to develop easy and rapid detection techniques for monitoring the implantation of inoculated O. oeni MLS on the indigenous LAB population, reducing the risk of unsuccessful MLF. PMID:20935116

Klebsiella pneumoniae type 3 fimbriae agglutinate yeast in a mannose-resistant manner.

PubMed

Stahlhut, Steen G; Struve, Carsten; Krogfelt, Karen A

2012-03-01

The ability of bacterial pathogens to express different fimbrial adhesins plays a significant role in virulence. Thus, specific detection of fimbrial expression is an important task in virulence characterization and epidemiological studies. Most clinical Klebsiella pneumoniae isolates express type 1 and type 3 fimbriae, which are characterized by mediation of mannose-sensitive agglutination of yeast cells and agglutination of tannic acid-treated ox red blood cells (RBCs), respectively. It has been observed that K. pneumoniae isolates agglutinate yeast cells and commercially available sheep RBCs in a mannose-resistant manner. Thus, this study was initiated to identify the adhesin involved. Screening of a mutant library surprisingly revealed that the mannose-resistant agglutination of yeast and sheep RBCs was mediated by type 3 fimbriae. Specific detection of type 1 fimbriae expression in K. pneumoniae was feasible only by the use of guinea pig RBCs. This was further verified by the use of isogenic fimbriae mutants and by cloning and expressing K. pneumoniae fimbrial gene clusters in Escherichia coli. Yeast agglutination assays are commonly used to detect type 1 fimbriae expression but should not be used for bacterial species able to express type 3 fimbriae. For these species, the use of guinea pig blood for specific type 1 fimbriae detection is essential. The use of commercially available sheep RBCs or yeast is an easy alternative to traditional methods to detect type 3 fimbriae expression. Easy and specific detection of expression of type 1 and type 3 fimbriae is essential in the continuous characterization of these important adhesive virulence factors present in members of the Enterobacteriaceae.

A material-sparing method for assessment of powder deformation characteristics using data collected during a single compression-decompression cycle.

PubMed

Katz, Jeffrey M; Roopwani, Rahul; Buckner, Ira S

2013-10-01

Compressibility profiles, or functions of solid fraction versus applied pressure, are used to provide insight into the fundamental mechanical behavior of powders during compaction. These functions, collected during compression (in-die) or post ejection (out-of-die), indicate the amount of pressure that a given powder formulation requires to be compressed to a given density or thickness. To take advantage of the benefits offered by both methods, the data collected in-die during a single compression-decompression cycle will be used to generate the equivalent of a complete out-of-die compressibility profile that has been corrected for both elastic and viscoelastic recovery of the powder. This method has been found to be both a precise and accurate means of evaluating out-of-die compressibility for four common tableting excipients. Using this method, a comprehensive characterization of powder compaction behavior, specifically in relation to plastic/brittle, elastic and viscoelastic deformation, can be obtained. Not only is the method computationally simple, but it is also material-sparing. The ability to characterize powder compressibility using this approach can improve productivity and streamline tablet development studies. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association.

[Receptor status (ER and PgR) determined with histochemical and biochemical methods in breast carcinoma].

PubMed

Simone, G; Paradiso, A; Cirillo, R; Mangia, A; Rella, G; Wiesel, S; Petroni, S; De Benedictis, G; De Lena, M

1991-01-01

Recently, a method similar to ER.ICA has been proposed for the progesterone receptor (PgR) using two monoclonal antibodies, JZB39 and KD68, specific for human PgR and characterized by a molecular weight of 95 and 120 Kd, respectively. A series of 73 breast cancer patients was studied with regards to ER and PgR using both immunocytochemical (ICA) and biochemical (DCC) assays. Results showed no substantial differences between the two methods when considering common clinical-pathological parameters. Overall agreement between ICA and DCC methods was found: 79% for PgR and 78% for ER. A slight quantitative correlation was also observed between the "score values" of the ICA method and the Fmol content of ER and PgR using the Brave-Pearson test (r = 0.49 for PgR; r = 0.43 for ER). Specificity of PgR.ICA method was 77% for PgR and 72% for ER; sensitivity was 82% and 83%, respectively. The ICA method is a reliable technique to assess PgR presence as well as ER. Further studies are necessary to evaluate the prognostic role of nuclear PgR.

Light scattering application for quantitative estimation of apoptosis

NASA Astrophysics Data System (ADS)

Bilyy, Rostyslav O.; Stoika, Rostyslav S.; Getman, Vasyl B.; Bilyi, Olexander I.

2004-05-01

Estimation of cell proliferation and apoptosis are in focus of instrumental methods used in modern biomedical sciences. Present study concerns monitoring of functional state of cells, specifically the development of their programmed death or apoptosis. The available methods for such purpose are either very expensive, or require time-consuming operations. Their specificity and sensitivity are frequently not sufficient for making conclusions which could be used in diagnostics or treatment monitoring. We propose a novel method for apoptosis measurement based on quantitative determination of cellular functional state taking into account their physical characteristics. This method uses the patented device -- laser microparticle analyser PRM-6 -- for analyzing light scattering by the microparticles, including cells. The method gives an opportunity for quick, quantitative, simple (without complicated preliminary cell processing) and relatively cheap measurement of apoptosis in cellular population. The elaborated method was used for studying apoptosis expression in murine leukemia cells of L1210 line and human lymphoblastic leukemia cells of K562 line. The results obtained by the proposed method permitted measuring cell number in tested sample, detecting and quantitative characterization of functional state of cells, particularly measuring the ratio of the apoptotic cells in suspension.

Identification and characterization of insect-specific proteins by genome data analysis

PubMed Central

Zhang, Guojie; Wang, Hongsheng; Shi, Junjie; Wang, Xiaoling; Zheng, Hongkun; Wong, Gane Ka-Shu; Clark, Terry; Wang, Wen; Wang, Jun; Kang, Le

2007-01-01

Background Insects constitute the vast majority of known species with their importance including biodiversity, agricultural, and human health concerns. It is likely that the successful adaptation of the Insecta clade depends on specific components in its proteome that give rise to specialized features. However, proteome determination is an intensive undertaking. Here we present results from a computational method that uses genome analysis to characterize insect and eukaryote proteomes as an approximation complementary to experimental approaches. Results Homologs in common to Drosophila melanogaster, Anopheles gambiae, Bombyx mori, Tribolium castaneum, and Apis mellifera were compared to the complete genomes of three non-insect eukaryotes (opisthokonts) Homo sapiens, Caenorhabditis elegans and Saccharomyces cerevisiae. This operation yielded 154 groups of orthologous proteins in Drosophila to be insect-specific homologs; 466 groups were determined to be common to eukaryotes (represented by three opisthokonts). ESTs from the hemimetabolous insect Locust migratoria were also considered in order to approximate their corresponding genes in the insect-specific homologs. Stress and stimulus response proteins were found to constitute a higher fraction in the insect-specific homologs than in the homologs common to eukaryotes. Conclusion The significant representation of stress response and stimulus response proteins in proteins determined to be insect-specific, along with specific cuticle and pheromone/odorant binding proteins, suggest that communication and adaptation to environments may distinguish insect evolution relative to other eukaryotes. The tendency for low Ka/Ks ratios in the insect-specific protein set suggests purifying selection pressure. The generally larger number of paralogs in the insect-specific proteins may indicate adaptation to environment changes. Instances in our insect-specific protein set have been arrived at through experiments reported in the literature, supporting the accuracy of our approach. PMID:17407609

MO-FG-CAMPUS-TeP1-05: Rapid and Efficient 3D Dosimetry for End-To-End Patient-Specific QA of Rotational SBRT Deliveries Using a High-Resolution EPID

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

Yang, Y M; Han, B; Xing, L

2016-06-15

Purpose: EPID-based patient-specific quality assurance provides verification of the planning setup and delivery process that phantomless QA and log-file based virtual dosimetry methods cannot achieve. We present a method for EPID-based QA utilizing spatially-variant EPID response kernels that allows for direct calculation of the entrance fluence and 3D phantom dose. Methods: An EPID dosimetry system was utilized for 3D dose reconstruction in a cylindrical phantom for the purposes of end-to-end QA. Monte Carlo (MC) methods were used to generate pixel-specific point-spread functions (PSFs) characterizing the spatially non-uniform EPID portal response in the presence of phantom scatter. The spatially-variant PSFs weremore » decomposed into spatially-invariant basis PSFs with the symmetric central-axis kernel as the primary basis kernel and off-axis representing orthogonal perturbations in pixel-space. This compact and accurate characterization enables the use of a modified Richardson-Lucy deconvolution algorithm to directly reconstruct entrance fluence from EPID images without iterative scatter subtraction. High-resolution phantom dose kernels were cogenerated in MC with the PSFs enabling direct recalculation of the resulting phantom dose by rapid forward convolution once the entrance fluence was calculated. A Delta4 QA phantom was used to validate the dose reconstructed in this approach. Results: The spatially-invariant representation of the EPID response accurately reproduced the entrance fluence with >99.5% fidelity with a simultaneous reduction of >60% in computational overhead. 3D dose for 10{sub 6} voxels was reconstructed for the entire phantom geometry. A 3D global gamma analysis demonstrated a >95% pass rate at 3%/3mm. Conclusion: Our approach demonstrates the capabilities of an EPID-based end-to-end QA methodology that is more efficient than traditional EPID dosimetry methods. Displacing the point of measurement external to the QA phantom reduces the necessary complexity of the phantom itself while offering a method that is highly scalable and inherently generalizable to rotational and trajectory based deliveries. This research was partially supported by Varian.« less

Nuclear morphology for the detection of alterations in bronchial cells from lung cancer: an attempt to improve sensitivity and specificity.

PubMed

Fafin-Lefevre, Mélanie; Morlais, Fabrice; Guittet, Lydia; Clin, Bénédicte; Launoy, Guy; Galateau-Sallé, Françoise; Plancoulaine, Benoît; Herlin, Paulette; Letourneux, Marc

2011-08-01

To identify which morphologic or densitometric parameters are modified in cell nuclei from bronchopulmonary cancer based on 18 parameters involving shape, intensity, chromatin, texture, and DNA content and develop a bronchopulmonary cancer screening method relying on analysis of sputum sample cell nuclei. A total of 25 sputum samples from controls and 22 bronchial aspiration samples from patients presenting with bronchopulmonary cancer who were professionally exposed to cancer were used. After Feulgen staining, 18 morphologic and DNA content parameters were measured on cell nuclei, via image cytom- etry. A method was developed for analyzing distribution quantiles, compared with simply interpreting mean values, to characterize morphologic modifications in cell nuclei. Distribution analysis of parameters enabled us to distinguish 13 of 18 parameters that demonstrated significant differences between controls and cancer cases. These parameters, used alone, enabled us to distinguish two population types, with both sensitivity and specificity > 70%. Three parameters offered 100% sensitivity and specificity. When mean values offered high sensitivity and specificity, comparable or higher sensitivity and specificity values were observed for at least one of the corresponding quantiles. Analysis of modification in morphologic parameters via distribution analysis proved promising for screening bronchopulmonary cancer from sputum.

Novel insight into antimicrobial resistance and sensitivity phenotypes associated to qac and norA genotypes in Staphylococcus aureus.

PubMed

Marchi, Emmanuela; Furi, Leonardo; Arioli, Stefania; Morrissey, Ian; Di Lorenzo, Valeria; Mora, Diego; Giovannetti, Luciana; Oggioni, Marco Rinaldo; Viti, Carlo

2015-01-01

Staphylococcus aureus strains harboring QacA, QacB, QacC, QacG transporters and norA promoter up-regulating mutations were characterized by phenotype microarray (PM), standard methods for susceptibility testing, and ethidium bromide efflux assays, in order to increase knowledge on phenotypes associated to efflux pumps and their substrates. PM data and standard susceptibility testing lead to the identification of new potential efflux targets, such as guanidine hydrochloride or 8-hydroxyquinoline for QacA and QacC pumps, respectively. The identification of compounds to which the presence of efflux pumps induced increased susceptibility opens new perspectives for potential adjunct anti-resistance treatment (i.e. strains bearing QacB transporters showed increased susceptibility to thioridazine, amitriptyline and orphenadrine). Although the tested isolates were characterized by high degree of heterogeneity, a hallmark of clinical isolates, direct ethidium bromide efflux assays were effective in highlighting differences in efflux efficiency among strains. These data add to characterization of substrate specificity in the different classes of staphylococcal multidrug efflux systems conferring specific substrate profiles and efflux features to each of them. Copyright © 2014 Elsevier GmbH. All rights reserved.

Towards a method to characterize temporary groundwater dynamics during droughts

NASA Astrophysics Data System (ADS)

Heudorfer, Benedikt; Stahl, Kerstin

2016-04-01

In order to improve our understanding of the complex mechanisms involved in the development, propagation and termination of drought events, a major challenge is to grasp the role of groundwater systems. Research on how groundwater responds to meteorological drought events (i.e. short-term climate anomalies) is still limited. Part of the problem is that there is as yet no generic method to characterize the response of different groundwater systems to extreme climate anomalies. In order to explore possibilities for such a methodology, we evaluate two statistical approaches to characterize groundwater dynamics on short time scales by applying them on observed groundwater head data from different pre- and peri-mountainous groundwater systems in humid central Europe (Germany). The first method is based on the coefficient of variation in moving windows of various lengths, the second method is based on streamflow recession characteristics applied on groundwater data. With these methods, the gauges behavior during low head events and its response to precipitation was explored. Findings regarding the behavior of the gauges make it possible to distinguish between gauges with a dominance of cyclic patterns, and gauges with a dominance of patterns on seasonal or event scale (commonly referred to as slow/fast responding gauges, respectively). While some clues on what factors that might control these patterns are present, the specific controls are general unclear for the gauges in this study. However as the key conclusion stands the question if the variety of manifestations of groundwater dynamics, as they occur in real systems, is subsumable with one unique method. Further studies on the topic are in progress.

Estimation of Staphylococcus aureus growth parameters from turbidity data: characterization of strain variation and comparison of methods.

PubMed

Lindqvist, R

2006-07-01

Turbidity methods offer possibilities for generating data required for addressing microorganism variability in risk modeling given that the results of these methods correspond to those of viable count methods. The objectives of this study were to identify the best approach for determining growth parameters based on turbidity data and use of a Bioscreen instrument and to characterize variability in growth parameters of 34 Staphylococcus aureus strains of different biotypes isolated from broiler carcasses. Growth parameters were estimated by fitting primary growth models to turbidity growth curves or to detection times of serially diluted cultures either directly or by using an analysis of variance (ANOVA) approach. The maximum specific growth rates in chicken broth at 17 degrees C estimated by time to detection methods were in good agreement with viable count estimates, whereas growth models (exponential and Richards) underestimated growth rates. Time to detection methods were selected for strain characterization. The variation of growth parameters among strains was best described by either the logistic or lognormal distribution, but definitive conclusions require a larger data set. The distribution of the physiological state parameter ranged from 0.01 to 0.92 and was not significantly different from a normal distribution. Strain variability was important, and the coefficient of variation of growth parameters was up to six times larger among strains than within strains. It is suggested to apply a time to detection (ANOVA) approach using turbidity measurements for convenient and accurate estimation of growth parameters. The results emphasize the need to consider implications of strain variability for predictive modeling and risk assessment.

Nano-zymography Using Laser-Scanning Confocal Microscopy Unmasks Proteolytic Activity of Cell-Derived Microparticles

PubMed Central

Briens, Aurélien; Gauberti, Maxime; Parcq, Jérôme; Montaner, Joan; Vivien, Denis; Martinez de Lizarrondo, Sara

2016-01-01

Cell-derived microparticles (MPs) are nano-sized vesicles released by activated cells in the extracellular milieu. They act as vectors of biological activity by carrying membrane-anchored and cytoplasmic constituents of the parental cells. Although detection and characterization of cell-derived MPs may be of high diagnostic and prognostic values in a number of human diseases, reliable measurement of their size, number and biological activity still remains challenging using currently available methods. In the present study, we developed a protocol to directly image and functionally characterize MPs using high-resolution laser-scanning confocal microscopy. Once trapped on annexin-V coated micro-wells, we developed several assays using fluorescent reporters to measure their size, detect membrane antigens and evaluate proteolytic activity (nano-zymography). In particular, we demonstrated the applicability and specificity of this method to detect antigens and proteolytic activities of tissue-type plasminogen activator (tPA), urokinase and plasmin at the surface of engineered MPs from transfected cell-lines. Furthermore, we were able to identify a subset of tPA-bearing fibrinolytic MPs using plasma samples from a cohort of ischemic stroke patients who received thrombolytic therapy and in an experimental model of thrombin-induced ischemic stroke in mice. Overall, this method is promising for functional characterization of cell-derived MPs. PMID:27022410

Nano-zymography Using Laser-Scanning Confocal Microscopy Unmasks Proteolytic Activity of Cell-Derived Microparticles.

PubMed

Briens, Aurélien; Gauberti, Maxime; Parcq, Jérôme; Montaner, Joan; Vivien, Denis; Martinez de Lizarrondo, Sara

2016-01-01

Cell-derived microparticles (MPs) are nano-sized vesicles released by activated cells in the extracellular milieu. They act as vectors of biological activity by carrying membrane-anchored and cytoplasmic constituents of the parental cells. Although detection and characterization of cell-derived MPs may be of high diagnostic and prognostic values in a number of human diseases, reliable measurement of their size, number and biological activity still remains challenging using currently available methods. In the present study, we developed a protocol to directly image and functionally characterize MPs using high-resolution laser-scanning confocal microscopy. Once trapped on annexin-V coated micro-wells, we developed several assays using fluorescent reporters to measure their size, detect membrane antigens and evaluate proteolytic activity (nano-zymography). In particular, we demonstrated the applicability and specificity of this method to detect antigens and proteolytic activities of tissue-type plasminogen activator (tPA), urokinase and plasmin at the surface of engineered MPs from transfected cell-lines. Furthermore, we were able to identify a subset of tPA-bearing fibrinolytic MPs using plasma samples from a cohort of ischemic stroke patients who received thrombolytic therapy and in an experimental model of thrombin-induced ischemic stroke in mice. Overall, this method is promising for functional characterization of cell-derived MPs.

Drug delivery properties of macroporous polystyrene solid foams.

PubMed

Canal, Cristina; Aparicio, Rosa Maria; Vilchez, Alejandro; Esquena, Jordi; García-Celma, Maria José

2012-01-01

Polymeric porous foams have been evaluated as possible new pharmaceutical dosage forms. These materials were obtained by polymerization in the continuous phase of highly concentrated emulsions prepared by the phase inversion temperature method. Their porosity, specific surface and surface topography were characterized, and the incorporation and release of active principles was studied using ketoprofen as model lipophilic molecule. Solid foams with very high pore volume, mainly inside macropores, were obtained by this method. The pore morphology of the materials was characterized, and very rough topography was observed, which contributed to their nearly superhydrophobic properties. These solid foams could be used as delivery systems for active principles with pharmaceutical interest, and in the present work ketoprofen was used as a model lipophilic molecule. Drug incorporation and release was studied from solid foam disks, using different concentrations of the loading solutions, achieving a delayed release with short lag-time.

Application of activity-based protein profiling to study enzyme function in adipocytes.

PubMed

Galmozzi, Andrea; Dominguez, Eduardo; Cravatt, Benjamin F; Saez, Enrique

2014-01-01

Activity-based protein profiling (ABPP) is a chemical proteomics approach that utilizes small-molecule probes to determine the functional state of enzymes directly in native systems. ABPP probes selectively label active enzymes, but not their inactive forms, facilitating the characterization of changes in enzyme activity that occur without alterations in protein levels. ABPP can be a tool superior to conventional gene expression and proteomic profiling methods to discover new enzymes active in adipocytes and to detect differences in the activity of characterized enzymes that may be associated with disorders of adipose tissue function. ABPP probes have been developed that react selectively with most members of specific enzyme classes. Here, using as an example the serine hydrolase family that includes many enzymes with critical roles in adipocyte physiology, we describe methods to apply ABPP analysis to the study of adipocyte enzymatic pathways. © 2014 Elsevier Inc. All rights reserved.

Characterization of suicidal behaviour with self-organizing maps.

PubMed

Leiva-Murillo, José M; López-Castromán, Jorge; Baca-García, Enrique

2013-01-01

The study of the variables involved in suicidal behavior is important from a social, medical, and economical point of view. Given the high number of potential variables of interest, a large population of subjects must be analysed in order to get conclusive results. In this paper, we describe a method based on self-organizing maps (SOMs) for finding the most relevant variables even when their relation to suicidal behavior is strongly nonlinear. We have applied the method to a cohort with more than 8,000 subjects and 600 variables and discovered four groups of variables involved in suicidal behavior. According to the results, there are four main groups of risk factors that characterize the population of suicide attempters: mental disorders, alcoholism, impulsivity, and childhood abuse. The identification of specific subpopulations of suicide attempters is consistent with current medical knowledge and may provide a new avenue of research to improve the management of suicidal cases.

Synthesis and characterization of Nafion/TiO2 nanocomposite membrane for proton exchange membrane fuel cell.

PubMed

Kim, Tae Young; Cho, Sung Yong

2011-08-01

In this study, the syntheses and characterizations of Nafion/TiO2 membranes for a proton exchange membrane fuel cell (PEMFC) were investigated. Porous TiO2 powders were synthesized using the sol-gel method; with Nafion/TiO2 nanocomposite membranes prepared using the casting method. An X-ray diffraction analysis demonstrated that the synthesized TiO2 had an anatase structure. The specific surface areas of the TiO2 and Nafion/TiO2 nanocomposite membrane were found to be 115.97 and 33.91 m2/g using a nitrogen adsorption analyzer. The energy dispersive spectra analysis indicated that the TiO2 particles were uniformly distributed in the nanocomposite membrane. The membrane electrode assembly prepared from the Nafion/TiO2 nanocomposite membrane gave the best PEMFC performance compared to the Nafion/P-25 and Nafion membranes.

Reprint of: Combining theory and experiment for X-ray absorption spectroscopy and resonant X-ray scattering characterization of polymers

DOE PAGES

Su, Gregory M.; Cordova, Isvar A.; Brady, Michael A.; ...

2016-11-01

An improved understanding of fundamental chemistry, electronic structure, morphology, and dynamics in polymers and soft materials requires advanced characterization techniques that are amenable to in situ and operando studies. Soft X-ray methods are especially useful in their ability to non-destructively provide information on specific materials or chemical moieties. Analysis of these experiments, which can be very dependent on X-ray energy and polarization, can quickly become complex. Complementary modeling and predictive capabilities are required to properly probe these critical features. Here in this paper, we present relevant background on this emerging suite of techniques. We focus on how the combination ofmore » theory and experiment has been applied and can be further developed to drive our understanding of how these methods probe relevant chemistry, structure, and dynamics in soft materials.« less

Combining theory and experiment for X-ray absorption spectroscopy and resonant X-ray scattering characterization of polymers

DOE PAGES

Su, Gregory M.; Cordova, Isvar A.; Brady, Michael A.; ...

2016-07-04

We present that an improved understanding of fundamental chemistry, electronic structure, morphology, and dynamics in polymers and soft materials requires advanced characterization techniques that are amenable to in situ and operando studies. Soft X-ray methods are especially useful in their ability to non-destructively provide information on specific materials or chemical moieties. Analysis of these experiments, which can be very dependent on X-ray energy and polarization, can quickly become complex. Complementary modeling and predictive capabilities are required to properly probe these critical features. Here, we present relevant background on this emerging suite of techniques. Finally, we focus on how the combinationmore » of theory and experiment has been applied and can be further developed to drive our understanding of how these methods probe relevant chemistry, structure, and dynamics in soft materials.« less

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Volume 6; Special Topics in Ocean Optics Protocols and Appendices; Revised

NASA Technical Reports Server (NTRS)

Mueller, J. L. (Editor); Fargion, Giulietta S. (Editor); McClain, Charles R. (Editor)

2003-01-01

This document stipulates protocols for measuring bio-optical and radiometric data for the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project activities and algorithm development. The document is organized into 6 separate volumes as Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 4. Volume I: Introduction, Background and Conventions; Volume II: Instrument Specifications, Characterization and Calibration; Volume III: Radiometric Measurements and Data Analysis Methods; Volume IV: Inherent Optical Properties: Instruments, Characterization, Field Measurements and Data Analysis Protocols; Volume V: Biogeochemical and Bio-Optical Measurements and Data Analysis Methods; Volume VI: Special Topics in Ocean Optics Protocols and Appendices. The earlier version of Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 3 (Mueller and Fargion 2002, Volumes 1 and 2) is entirely superseded by the six volumes of Revision 4 listed above.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding.

PubMed

Shahi, Payam; Kim, Samuel C; Haliburton, John R; Gartner, Zev J; Abate, Adam R

2017-03-14

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

NASA Astrophysics Data System (ADS)

Shahi, Payam; Kim, Samuel C.; Haliburton, John R.; Gartner, Zev J.; Abate, Adam R.

2017-03-01

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

PubMed Central

Shahi, Payam; Kim, Samuel C.; Haliburton, John R.; Gartner, Zev J.; Abate, Adam R.

2017-01-01

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing. PMID:28290550

Correlation Between Hot Spots and 3-d Defect Structure in Single and Polycrystalline High-explosive Materials

NASA Astrophysics Data System (ADS)

Hawkins, Cameron; Tschuaner, Oliver; Fussell, Zachary; Smith, Jesse

2017-06-01

A novel approach that spatially identifies inhomogeneities from microscale (defects, con-formational disorder) to mesoscale (voids, inclusions) is developed using synchrotron x-ray methods: tomography, Lang topography, and micro-diffraction mapping. These techniques pro-vide a non-destructive method for characterization of mm-sized samples prior to shock experiments. These characterization maps can be used to correlate continuum level measurements in shock compression experiments to the mesoscale and microscale structure. Specifically examined is a sample of C4. We show extensive conformational disorder in gamma-RDX, which is the main component. Further, we observe that the minor HMX-component in C4 contains at least two different phases: alpha- and beta-HMX. This work supported by National Security Technologies, LLC, under Contract No. DE-AC52-06NA25946 with the U.S. Department of Energy and by the Site-Directed Research and Development Program. DOE/NV/25946-3071.

Recombinant G protein-coupled receptor expression in Saccharomyces cerevisiae for protein characterization.

PubMed

Blocker, Kory M; Britton, Zachary T; Naranjo, Andrea N; McNeely, Patrick M; Young, Carissa L; Robinson, Anne S

2015-01-01

G protein-coupled receptors (GPCRs) are membrane proteins that mediate signaling across the cellular membrane and facilitate cellular responses to external stimuli. Due to the critical role that GPCRs play in signal transduction, therapeutics have been developed to influence GPCR function without an extensive understanding of the receptors themselves. Closing this knowledge gap is of paramount importance to improving therapeutic efficacy and specificity, where efforts to achieve this end have focused chiefly on improving our knowledge of the structure-function relationship. The purpose of this chapter is to review methods for the heterologous expression of GPCRs in Saccharomyces cerevisiae, including whole-cell assays that enable quantitation of expression, localization, and function in vivo. In addition, we describe methods for the micellular solubilization of the human adenosine A2a receptor and for reconstitution of the receptor in liposomes that have enabled its biophysical characterization. © 2015 Elsevier Inc. All rights reserved.

Protein vivisection reveals elusive intermediates in folding

PubMed Central

Zheng, Zhongzhou; Sosnick, Tobin R.

2010-01-01

Although most folding intermediates escape detection, their characterization is crucial to the elucidation of folding mechanisms. Here we outline a powerful strategy to populate partially unfolded intermediates: A buried aliphatic residue is substituted with a charged residue (e.g., Leu→Glu−) to destabilize and unfold a specific region of the protein. We apply this strategy to Ubiquitin, reversibly trapping a folding intermediate in which the β5 strand is unfolded. The intermediate refolds to a native-like structure upon charge neutralization under mildly acidic conditions. Characterization of the trapped intermediate using NMR and hydrogen exchange methods identifies a second folding intermediate and reveals the order and free energies of the two major folding events on the native side of the rate-limiting step. This general strategy may be combined with other methods and have broad applications in the study of protein folding and other reactions that require trapping of high energy states. PMID:20144618

Characterization of Arabidopsis thaliana mismatch specific endonucleases: application to mutation discovery by TILLING in pea.

PubMed

Triques, Karine; Sturbois, Bénédicte; Gallais, Stéphane; Dalmais, Marion; Chauvin, Stéphanie; Clepet, Christian; Aubourg, Sébastien; Rameau, Catherine; Caboche, Michel; Bendahmane, Abdelhafid

2007-09-01

Scanning DNA sequences for mutations and polymorphisms has become one of the most challenging, often expensive and time-consuming obstacles in many molecular genetic applications, including reverse genetic and clinical diagnostic applications. Enzymatic mutation detection methods are based on the cleavage of heteroduplex DNA at the mismatch sites. These methods are often limited by the availability of a mismatch-specific endonuclease, their sensitivity in detecting one allele in a pool of DNA and their costs. Here, we present detailed biochemical analysis of five Arabidopsis putative mismatch-specific endonucleases. One of them, ENDO1, is presented as the first endonuclease that recognizes and cleaves all types of mismatches with high efficiency. We report on a very simple protocol for the expression and purification of ENDO1. The ENDO1 system could be exploited in a wide range of mutation diagnostic tools. In particular, we report the use of ENDO1 for discovery of point mutations in the gibberellin 3beta-hydrolase gene of Pisum sativum. Twenty-one independent mutants were isolated, five of these were characterized and two new mutations affecting internodes length were identified. To further evaluate the quality of the mutant population we screened for mutations in four other genes and identified 5-21 new alleles per target. Based on the frequency of the obtained alleles we concluded that the pea population described here would be suitable for use in a large reverse-genetics project.

Detection of low-level environmental chemical allergy by a long-term sensitization method.

PubMed

Fukuyama, Tomoki; Ueda, Hideo; Hayashi, Koichi; Tajima, Yukari; Shuto, Yasufumi; Saito, Toru R; Harada, Takanori; Kosaka, Tadashi

2008-07-30

Multiple chemical sensitivity (MCS) is characterized by various signs, including neurological disorders and allergy. Exposure may occur through a major event, such as a chemical spill, or from long-term contact with chemicals at low levels. We are interested in the allergenicity of MCS and the detection of low-level chemical-related hypersensitivity. We used long-term sensitization followed by low-dose challenge to evaluate sensitization by well-known Th2 type sensitizers (trimellitic anhydride (TMA) and toluene diisocyanate (TDI)) and a Th1 type sensitizer (2,4-dinitrochlorobenzene (DNCB)). After topically sensitizing BALB/c mice (9 times in 3 weeks) and challenging them with TMA, TDI or DNCB, we assayed their auricular lymph nodes (LNs) for number of lymphocytes, surface antigen expression of B cells, and local cytokine production, and measured antigen-specific serum IgE levels. TMA and TDI induced marked increases in levels of antigen-specific serum IgE and of Th2 cytokines (IL-4, IL-5, IL-10, and IL-13) produced by ex vivo restimulated lymph node cells. DNCB induced a marked increase in Th1 cytokine (IL-2, IFN-gamma, and TNF-alpha) levels, but antigen-specific serum IgE levels were not elevated. All chemicals induced significant increases in number of lymphocytes and surface antigen expression of B cells. Our mouse model enabled the identification and characterization of chemical-related allergic reactions at low levels. This long-term sensitization method would be useful for detecting environmental chemical-related hypersensitivity.

Practical utilization of recombinant AAV vector reference standards: focus on vector genomes titration by free ITR qPCR.

PubMed

D'Costa, Susan; Blouin, Veronique; Broucque, Frederic; Penaud-Budloo, Magalie; François, Achille; Perez, Irene C; Le Bec, Christine; Moullier, Philippe; Snyder, Richard O; Ayuso, Eduard

2016-01-01

Clinical trials using recombinant adeno-associated virus (rAAV) vectors have demonstrated efficacy and a good safety profile. Although the field is advancing quickly, vector analytics and harmonization of dosage units are still a limitation for commercialization. AAV reference standard materials (RSMs) can help ensure product safety by controlling the consistency of assays used to characterize rAAV stocks. The most widely utilized unit of vector dosing is based on the encapsidated vector genome. Quantitative polymerase chain reaction (qPCR) is now the most common method to titer vector genomes (vg); however, significant inter- and intralaboratory variations have been documented using this technique. Here, RSMs and rAAV stocks were titered on the basis of an inverted terminal repeats (ITRs) sequence-specific qPCR and we found an artificial increase in vg titers using a widely utilized approach. The PCR error was introduced by using single-cut linearized plasmid as the standard curve. This bias was eliminated using plasmid standards linearized just outside the ITR region on each end to facilitate the melting of the palindromic ITR sequences during PCR. This new "Free-ITR" qPCR delivers vg titers that are consistent with titers obtained with transgene-specific qPCR and could be used to normalize in-house product-specific AAV vector standards and controls to the rAAV RSMs. The free-ITR method, including well-characterized controls, will help to calibrate doses to compare preclinical and clinical data in the field.

A method for detecting and characterizing outbreaks of infectious disease from clinical reports.

PubMed

Cooper, Gregory F; Villamarin, Ricardo; Rich Tsui, Fu-Chiang; Millett, Nicholas; Espino, Jeremy U; Wagner, Michael M

2015-02-01

Outbreaks of infectious disease can pose a significant threat to human health. Thus, detecting and characterizing outbreaks quickly and accurately remains an important problem. This paper describes a Bayesian framework that links clinical diagnosis of individuals in a population to epidemiological modeling of disease outbreaks in the population. Computer-based diagnosis of individuals who seek healthcare is used to guide the search for epidemiological models of population disease that explain the pattern of diagnoses well. We applied this framework to develop a system that detects influenza outbreaks from emergency department (ED) reports. The system diagnoses influenza in individuals probabilistically from evidence in ED reports that are extracted using natural language processing. These diagnoses guide the search for epidemiological models of influenza that explain the pattern of diagnoses well. Those epidemiological models with a high posterior probability determine the most likely outbreaks of specific diseases; the models are also used to characterize properties of an outbreak, such as its expected peak day and estimated size. We evaluated the method using both simulated data and data from a real influenza outbreak. The results provide support that the approach can detect and characterize outbreaks early and well enough to be valuable. We describe several extensions to the approach that appear promising. Copyright © 2014 Elsevier Inc. All rights reserved.

A Method for Detecting and Characterizing Outbreaks of Infectious Disease from Clinical Reports

PubMed Central

Cooper, Gregory F.; Villamarin, Ricardo; Tsui, Fu-Chiang (Rich); Millett, Nicholas; Espino, Jeremy U.; Wagner, Michael M.

2014-01-01

Outbreaks of infectious disease can pose a significant threat to human health. Thus, detecting and characterizing outbreaks quickly and accurately remains an important problem. This paper describes a Bayesian framework that links clinical diagnosis of individuals in a population to epidemiological modeling of disease outbreaks in the population. Computer-based diagnosis of individuals who seek healthcare is used to guide the search for epidemiological models of population disease that explain the pattern of diagnoses well. We applied this framework to develop a system that detects influenza outbreaks from emergency department (ED) reports. The system diagnoses influenza in individuals probabilistically from evidence in ED reports that are extracted using natural language processing. These diagnoses guide the search for epidemiological models of influenza that explain the pattern of diagnoses well. Those epidemiological models with a high posterior probability determine the most likely outbreaks of specific diseases; the models are also used to characterize properties of an outbreak, such as its expected peak day and estimated size. We evaluated the method using both simulated data and data from a real influenza outbreak. The results provide support that the approach can detect and characterize outbreaks early and well enough to be valuable. We describe several extensions to the approach that appear promising. PMID:25181466

De novo assembly of the transcriptome of the non-model plant Streptocarpus rexii employing a novel heuristic to recover locus-specific transcript clusters.

PubMed

Chiara, Matteo; Horner, David S; Spada, Alberto

2013-01-01

De novo transcriptome characterization from Next Generation Sequencing data has become an important approach in the study of non-model plants. Despite notable advances in the assembly of short reads, the clustering of transcripts into unigene-like (locus-specific) clusters remains a somewhat neglected subject. Indeed, closely related paralogous transcripts are often merged into single clusters by current approaches. Here, a novel heuristic method for locus-specific clustering is compared to that implemented in the de novo assembler Oases, using the same initial transcript collections, derived from Arabidopsis thaliana and the developmental model Streptocarpus rexii. We show that the proposed approach improves cluster specificity in the A. thaliana dataset for which the reference genome is available. Furthermore, for the S. rexii data our filtered transcript collection matches a larger number of distinct annotated loci in reference genomes than the Oases set, while containing a reduced overall number of loci. A detailed discussion of advantages and limitations of our approach in processing de novo transcriptome reconstructions is presented. The proposed method should be widely applicable to other organisms, irrespective of the transcript assembly method employed. The S. rexii transcriptome is available as a sophisticated and augmented publicly available online database.

Visualization of genome signatures of eukaryote genomes by batch-learning self-organizing map with a special emphasis on Drosophila genomes.

PubMed

Abe, Takashi; Hamano, Yuta; Ikemura, Toshimichi

2014-01-01

A strategy of evolutionary studies that can compare vast numbers of genome sequences is becoming increasingly important with the remarkable progress of high-throughput DNA sequencing methods. We previously established a sequence alignment-free clustering method "BLSOM" for di-, tri-, and tetranucleotide compositions in genome sequences, which can characterize sequence characteristics (genome signatures) of a wide range of species. In the present study, we generated BLSOMs for tetra- and pentanucleotide compositions in approximately one million sequence fragments derived from 101 eukaryotes, for which almost complete genome sequences were available. BLSOM recognized phylotype-specific characteristics (e.g., key combinations of oligonucleotide frequencies) in the genome sequences, permitting phylotype-specific clustering of the sequences without any information regarding the species. In our detailed examination of 12 Drosophila species, the correlation between their phylogenetic classification and the classification on the BLSOMs was observed to visualize oligonucleotides diagnostic for species-specific clustering.

Development of an enzyme-linked-immunosorbent-assay technique for accurate identification of poorly preserved silks unearthed in ancient tombs.

PubMed

Zheng, Qin; Wu, Xiaofeng; Zheng, Hailing; Zhou, Yang

2015-05-01

We report the preparation of a specific fibroin antibody and its use for the identification of unearthed ancient silk relics. Based on the 12-amino-acid repeat sequence "GAGAGSGAGAGS", which is found in fibroin of the silkworm Bombyx mori, a specific antibody against fibroin was prepared in rabbits through peptide synthesis and carrier-protein coupling. This antibody was highly specific for fibroin found in silk. Using this antibody we have successfully identified four silk samples from different time periods. Our results reveal, for the first time, a method capable of detecting silk from a few milligrams of archaeological fabric that has been buried for thousands of years, confirming that the ancient practice of wearing silk products while praying for rebirth dated back to at least 400 BCE. This method also complements current approaches in silk detection, especially for the characterization of poorly preserved silks, promoting the investigation of silk origins and of ancient clothing cultures.

Optimization and characterization of bioactive glass nanofibers and nanocomposites

NASA Astrophysics Data System (ADS)

Scarber, Reginna E.

Disease affects different areas of the bone and can impact individuals of all pathologies and ethnicities. These bone diseases can result in weakening which leads to trauma during ordinary function, the need for reconstructive surgery, and eventual bone replacement. Tissue engineering can provide a less traumatic and more fundamental solution to the current therapies. Bioactive glasses are promising materials in tissue engineering applications because of their ability to form hydroxycarbonate apatite in the presence of simulated body fluid, support cell adhesion, growth, and differentiation, induce bone formation, and concentrate bone morphogenic proteins in vivo. The research in this dissertation will attempt to improve the quality, yield, and toughness of bioactive glass nanofibrous scaffolds. The three specific aims of this research include, (1) Optimization and Characterization of Surfactant Modified Bioactive Glass (2) Optimization of Direct Synthesis Bioactive glass Nanofibers from Sols (3) Mechanical Properties and In-vitro Biomineralization of Bioglass-loaded Polyglyconate Nanocomposites Created Using the Particulate Leaching Method. The purpose of the first specific aim was to optimize the processing of bioactive glass nanofibers, resulting in greater fiber uniformity with a reduction in beading. The increase in viscosity coupled with the ability of the surfactant to limit polymeric secondary bonding led to improved fiber quality. The focal point of the second specific aim is the production of sol-gel derived glass fibers with high bioactivity prepared by electrospinning without the use of any polymer carrier system. Advantages of this method include decreased processing time, increased production of fibers, and a decrease in the loss of material due to the calcining process. The solvent cast/ particulate leaching method was used to create a nanocomposite of bioglass and the co-polymer polyglyconate (MaxonRTM) for bone tissue scaffolds The biocompatibility of the composite foams was observed and calcium phosphate presence was quantified. The incorporation of bioglass into the polymer matrix improved the strength (modulus - 21.47 MPa) and biocompatibility of the polyglyconate foam. Keywords: Bioactive glass, Electrospinning, Solvent Casting/Particulate Leaching Method, Nanocomposites

History, rare, and multiple events of mechanical unfolding of repeat proteins

NASA Astrophysics Data System (ADS)

Sumbul, Fidan; Marchesi, Arin; Rico, Felix

2018-03-01

Mechanical unfolding of proteins consisting of repeat domains is an excellent tool to obtain large statistics. Force spectroscopy experiments using atomic force microscopy on proteins presenting multiple domains have revealed that unfolding forces depend on the number of folded domains (history) and have reported intermediate states and rare events. However, the common use of unspecific attachment approaches to pull the protein of interest holds important limitations to study unfolding history and may lead to discarding rare and multiple probing events due to the presence of unspecific adhesion and uncertainty on the pulling site. Site-specific methods that have recently emerged minimize this uncertainty and would be excellent tools to probe unfolding history and rare events. However, detailed characterization of these approaches is required to identify their advantages and limitations. Here, we characterize a site-specific binding approach based on the ultrastable complex dockerin/cohesin III revealing its advantages and limitations to assess the unfolding history and to investigate rare and multiple events during the unfolding of repeated domains. We show that this approach is more robust, reproducible, and provides larger statistics than conventional unspecific methods. We show that the method is optimal to reveal the history of unfolding from the very first domain and to detect rare events, while being more limited to assess intermediate states. Finally, we quantify the forces required to unfold two molecules pulled in parallel, difficult when using unspecific approaches. The proposed method represents a step forward toward more reproducible measurements to probe protein unfolding history and opens the door to systematic probing of rare and multiple molecule unfolding mechanisms.

RNA Flow Cytometry Using the Branched DNA Technique.

PubMed

Soh, Kah Teong; Wallace, Paul K

2018-01-01

The systematic modulation of mRNA and proteins governs the complicated and intermingled biological functions of our cells. Traditionally, transcriptomic technologies such as DNA microarray and RNA-Seq have been used to identify, characterize, and profile gene expression data. These are, however, considered bulk methods as they are unable to measure gene expression at the single-cell level, unless the cells are pre-sorted. Branched DNA is a flow cytometry-based detection platform that has been developed recently to measure mRNA at the single-cell level. Originally adapted from microscopy, the current system has been modified to achieve compatibility with the detection of surface and intracellular antigens using monoclonal antibodies conjugated to fluorochromes, thus permitting simultaneous detection of mRNAs and proteins. The Branched DNA method offers a variety of advantages when compared to traditional or standard methods used for the quantification of mRNA, such as (a) the detection of specific mRNA on a per cell basis, (b) an alternate detection tool when the measurement of a protein is technically infeasible (i.e., no quality antibody exists) or the epitope is not assessable, and (c) correlate the analysis of mRNA with protein. Compared to earlier attempts at measuring nucleic acid by flow cytometry, the hybridization temperature applied in the Branched DNA assay is much lower, thus preserving the integrity of cellular structures for further characterization. It also has greatly increased specificity and sensitivity. Here, we provide detailed instruction for performing the Branched DNA method using it in a model system to correlate the expression of CD8 mRNA and CD8 protein by flow cytometry.

Novel characterization of the aerosol and gas-phase composition of aerosolized jet fuel.

PubMed

Tremblay, Raphael T; Martin, Sheppard A; Fisher, Jeffrey W

2010-04-01

Few robust methods are available to characterize the composition of aerosolized complex hydrocarbon mixtures. The difficulty in separating the droplets from their surrounding vapors and preserving their content is challenging, more so with fuels, which contain hydrocarbons ranging from very low to very high volatility. Presented here is a novel method that uses commercially available absorbent tubes to measure a series of hydrocarbons in the vapor and droplets from aerosolized jet fuels. Aerosol composition and concentrations were calculated from the differential between measured total (aerosol and gas-phase) and measured gas-phase concentrations. Total samples were collected directly, whereas gas-phase only samples were collected behind a glass fiber filter to remove droplets. All samples were collected for 1 min at 400 ml min(-1) and quantified using thermal desorption-gas chromatography-mass spectrometry. This method was validated for the quantification of the vapor and droplet content from 4-h aerosolized jet fuel exposure to JP-8 and S-8 at total concentrations ranging from 200 to 1000 mg/m(3). Paired samples (gas-phase only and total) were collected every approximately 40 min. Calibrations were performed with neat fuel to calculate total concentration and also with a series of authentic standards to calculate specific compound concentrations. Accuracy was good when compared to an online GC-FID (gas chromatography-flame ionization detection) technique. Variability was 15% or less for total concentrations, the sum of all gas-phase compounds, and for most specific compound concentrations in both phases. Although validated for jet fuels, this method can be adapted to other hydrocarbon-based mixtures.

Dynamics of Dengue Virus (DENV)–Specific B Cells in the Response to DENV Serotype 1 Infections, Using Flow Cytometry With Labeled Virions

PubMed Central

Woda, Marcia; Friberg, Heather; Currier, Jeffrey R.; Srikiatkhachorn, Anon; Macareo, Louis R.; Green, Sharone; Jarman, Richard G.; Rothman, Alan L.; Mathew, Anuja

2016-01-01

Background. The development of reagents to identify and characterize antigen-specific B cells has been challenging. Methods. We recently developed Alexa Fluor–labeled dengue viruses (AF DENVs) to characterize antigen-specific B cells in the peripheral blood of DENV-immune individuals. Results. In this study, we used AF DENV serotype 1 (AF DENV-1) together with AF DENV-2 on peripheral blood mononuclear cells (PBMCs) from children in Thailand with acute primary or secondary DENV-1 infections to analyze the phenotypes of antigen-specific B cells that reflected their exposure or clinical diagnosis. DENV serotype-specific and cross-reactive B cells were identified in PBMCs from all subjects. Frequencies of AF DENV+ class-switched memory B cells (IgD−CD27+ CD19+ cells) reached up to 8% during acute infection and early convalescence. AF DENV–labeled B cells expressed high levels of CD27 and CD38 during acute infection, characteristic of plasmablasts, and transitioned into memory B cells (CD38−CD27+) at the early convalescent time point. There was higher activation of memory B cells early during acute secondary infection, suggesting reactivation from a previous DENV infection. Conclusions. AF DENVs reveal changes in the phenotype of DENV serotype–specific and cross-reactive B cells during and after natural DENV infection and could be useful in analysis of the response to DENV vaccination. PMID:27443614

Isolation of HIV-1-Neutralizing Mucosal Monoclonal Antibodies from Human Colostrum

PubMed Central

Friedman, James; Alam, S. Munir; Shen, Xiaoying; Xia, Shi-Mao; Stewart, Shelley; Anasti, Kara; Pollara, Justin; Fouda, Genevieve G.; Yang, Guang; Kelsoe, Garnett; Ferrari, Guido; Tomaras, Georgia D.; Haynes, Barton F.; Liao, Hua-Xin

2012-01-01

Background Generation of potent anti-HIV antibody responses in mucosal compartments is a potential requirement of a transmission-blocking HIV vaccine. HIV-specific, functional antibody responses are present in breast milk, and these mucosal antibody responses may play a role in protection of the majority of HIV-exposed, breastfeeding infants. Therefore, characterization of HIV-specific antibodies produced by B cells in milk could guide the development of vaccines that elicit protective mucosal antibody responses. Methods We isolated B cells from colostrum of an HIV-infected lactating woman with a detectable neutralization response in milk and recombinantly produced and characterized the resulting HIV-1 Envelope (Env)-specific monoclonal antibodies (mAbs). Results The identified HIV-1 Env-specific colostrum mAbs, CH07 and CH08, represent two of the first mucosally-derived anti-HIV antibodies yet to be reported. Colostrum mAb CH07 is a highly-autoreactive, weakly-neutralizing gp140-specific mAb that binds to linear epitopes in the gp120 C5 region and gp41 fusion domain. In contrast, colostrum mAb CH08 is a nonpolyreactive CD4-inducible (CD4i) gp120-specific mAb with moderate breadth of neutralization. Conclusions These novel HIV-neutralizing mAbs isolated from a mucosal compartment provide insight into the ability of mucosal B cell populations to produce functional anti-HIV antibodies that may contribute to protection against virus acquisition at mucosal surfaces. PMID:22624058

Peptide Probe for Crystalline Hydroxyapatite: In Situ Detection of Biomineralization

NASA Astrophysics Data System (ADS)

Cicerone, Marcus; Becker, Matthew; Simon, Carl; Chatterjee, Kaushik

2009-03-01

While cells template mineralization in vitro and in vivo, specific detection strategies that impart chemical and structural information on this process have proven elusive. Recently we have developed an in situ based peptide probe via phage display methods that is specific to crystalline hydroxyapatite (HA). We are using this in fluorescence based assays to characterize mineralization. One application being explored is the screening of tissue engineering scaffolds for their ability to support osteogenesis. Specifically, osteoblasts are being cultured in hydrogel scaffolds possessing property gradients to provide a test bed for the HA peptide probe. Hydrogel properties that support osteogenesis and HA deposition will be identified using the probe to demonstrate its utility in optimizing design of tissue scaffolds.

A statistical method for estimating rates of soil development and ages of geologic deposits: A design for soil-chronosequence studies

USGS Publications Warehouse

Switzer, P.; Harden, J.W.; Mark, R.K.

1988-01-01

A statistical method for estimating rates of soil development in a given region based on calibration from a series of dated soils is used to estimate ages of soils in the same region that are not dated directly. The method is designed specifically to account for sampling procedures and uncertainties that are inherent in soil studies. Soil variation and measurement error, uncertainties in calibration dates and their relation to the age of the soil, and the limited number of dated soils are all considered. Maximum likelihood (ML) is employed to estimate a parametric linear calibration curve, relating soil development to time or age on suitably transformed scales. Soil variation on a geomorphic surface of a certain age is characterized by replicate sampling of soils on each surface; such variation is assumed to have a Gaussian distribution. The age of a geomorphic surface is described by older and younger bounds. This technique allows age uncertainty to be characterized by either a Gaussian distribution or by a triangular distribution using minimum, best-estimate, and maximum ages. The calibration curve is taken to be linear after suitable (in certain cases logarithmic) transformations, if required, of the soil parameter and age variables. Soil variability, measurement error, and departures from linearity are described in a combined fashion using Gaussian distributions with variances particular to each sampled geomorphic surface and the number of sample replicates. Uncertainty in age of a geomorphic surface used for calibration is described using three parameters by one of two methods. In the first method, upper and lower ages are specified together with a coverage probability; this specification is converted to a Gaussian distribution with the appropriate mean and variance. In the second method, "absolute" older and younger ages are specified together with a most probable age; this specification is converted to an asymmetric triangular distribution with mode at the most probable age. The statistical variability of the ML-estimated calibration curve is assessed by a Monte Carlo method in which simulated data sets repeatedly are drawn from the distributional specification; calibration parameters are reestimated for each such simulation in order to assess their statistical variability. Several examples are used for illustration. The age of undated soils in a related setting may be estimated from the soil data using the fitted calibration curve. A second simulation to assess age estimate variability is described and applied to the examples. ?? 1988 International Association for Mathematical Geology.

Fine epitope signature of antibody neutralization breadth at the HIV-1 envelope CD4-binding site.

PubMed

Cheng, Hao D; Grimm, Sebastian K; Gilman, Morgan Sa; Gwom, Luc Christian; Sok, Devin; Sundling, Christopher; Donofrio, Gina; Hedestam, Gunilla B Karlsson; Bonsignori, Mattia; Haynes, Barton F; Lahey, Timothy P; Maro, Isaac; von Reyn, C Fordham; Gorny, Miroslaw K; Zolla-Pazner, Susan; Walker, Bruce D; Alter, Galit; Burton, Dennis R; Robb, Merlin L; Krebs, Shelly J; Seaman, Michael S; Bailey-Kellogg, Chris; Ackerman, Margaret E

2018-03-08

Major advances in donor identification, antigen probe design, and experimental methods to clone pathogen-specific antibodies have led to an exponential growth in the number of newly characterized broadly neutralizing antibodies (bnAbs) that recognize the HIV-1 envelope glycoprotein. Characterization of these bnAbs has defined new epitopes and novel modes of recognition that can result in potent neutralization of HIV-1. However, the translation of envelope recognition profiles in biophysical assays into an understanding of in vivo activity has lagged behind, and identification of subjects and mAbs with potent antiviral activity has remained reliant on empirical evaluation of neutralization potency and breadth. To begin to address this discrepancy between recombinant protein recognition and virus neutralization, we studied the fine epitope specificity of a panel of CD4-binding site (CD4bs) antibodies to define the molecular recognition features of functionally potent humoral responses targeting the HIV-1 envelope site bound by CD4. Whereas previous studies have used neutralization data and machine-learning methods to provide epitope maps, here, this approach was reversed, demonstrating that simple binding assays of fine epitope specificity can prospectively identify broadly neutralizing CD4bs-specific mAbs. Building on this result, we show that epitope mapping and prediction of neutralization breadth can also be accomplished in the assessment of polyclonal serum responses. Thus, this study identifies a set of CD4bs bnAb signature amino acid residues and demonstrates that sensitivity to mutations at signature positions is sufficient to predict neutralization breadth of polyclonal sera with a high degree of accuracy across cohorts and across clades.

"Hook"-calibration of GeneChip-microarrays: theory and algorithm.

PubMed

Binder, Hans; Preibisch, Stephan

2008-08-29

: The improvement of microarray calibration methods is an essential prerequisite for quantitative expression analysis. This issue requires the formulation of an appropriate model describing the basic relationship between the probe intensity and the specific transcript concentration in a complex environment of competing interactions, the estimation of the magnitude these effects and their correction using the intensity information of a given chip and, finally the development of practicable algorithms which judge the quality of a particular hybridization and estimate the expression degree from the intensity values. : We present the so-called hook-calibration method which co-processes the log-difference (delta) and -sum (sigma) of the perfect match (PM) and mismatch (MM) probe-intensities. The MM probes are utilized as an internal reference which is subjected to the same hybridization law as the PM, however with modified characteristics. After sequence-specific affinity correction the method fits the Langmuir-adsorption model to the smoothed delta-versus-sigma plot. The geometrical dimensions of this so-called hook-curve characterize the particular hybridization in terms of simple geometric parameters which provide information about the mean non-specific background intensity, the saturation value, the mean PM/MM-sensitivity gain and the fraction of absent probes. This graphical summary spans a metrics system for expression estimates in natural units such as the mean binding constants and the occupancy of the probe spots. The method is single-chip based, i.e. it separately uses the intensities for each selected chip. : The hook-method corrects the raw intensities for the non-specific background hybridization in a sequence-specific manner, for the potential saturation of the probe-spots with bound transcripts and for the sequence-specific binding of specific transcripts. The obtained chip characteristics in combination with the sensitivity corrected probe-intensity values provide expression estimates scaled in natural units which are given by the binding constants of the particular hybridization.

Characterization of Infrastructure Materials using Nonlinear Ultrasonics

NASA Astrophysics Data System (ADS)

Liu, Minghe

In order to improve the safety, reliability, cost, and performance of civil and mechanical structures/components, it is necessary to develop techniques that are capable of characterizing and quantifying the amount of distributed damage in engineering materials before any detectable discontinuities (cracks, delaminations, voids, etc.) appear. In this dissertation, novel nonlinear ultrasonic NDE methods are developed and applied to characterize cumulative damage such as fatigue damage in metallic materials and degradation of cement-based materials due to chemical reactions. First, nonlinear Rayleigh surface waves are used to measure the near-surface residual stresses in shot-peened aluminum alloy (AA 7075) samples. Results show that the nonlinear Rayleigh wave is very sensitive to near-surface residual stresses, and has the potential to quantitatively detect them. Second, a novel two-wave mixing method is theoretically developed and numerically verified. This method is then successfully applied to detect the fatigue damage in aluminum alloy (AA 6061) samples subjected to monotonic compression. In addition to its high sensitivity to fatigue damage, this collinear wave mixing method allows the measurement over a specific region of interest in the specimen, and this capability makes it possible to obtain spatial distribution of fatigue damage through the thickness direction of the sample by simply timing the transducers. Third, the nonlinear wave mixing method is used to characterize the degradation of cement-based materials caused by alkali-silica reaction (ASR). It is found that the nonlinear ultrasonic method is sensitive to detect ASR damage at very early stage, and has the potential to identify the different damage stages. Finally, a micromechanics-based chemo-mechanical model is developed which relates the acoustic nonlinearity parameter to ASR damage. This model provides a way to quantitatively predict the changes in the acoustic nonlinearity parameter due to ASR damage, which can be used to guide experimental measurements for nondestructive evaluation of ASR damage.

Texture-specific bag of visual words model and spatial cone matching-based method for the retrieval of focal liver lesions using multiphase contrast-enhanced CT images.

PubMed

Xu, Yingying; Lin, Lanfen; Hu, Hongjie; Wang, Dan; Zhu, Wenchao; Wang, Jian; Han, Xian-Hua; Chen, Yen-Wei

2018-01-01

The bag of visual words (BoVW) model is a powerful tool for feature representation that can integrate various handcrafted features like intensity, texture, and spatial information. In this paper, we propose a novel BoVW-based method that incorporates texture and spatial information for the content-based image retrieval to assist radiologists in clinical diagnosis. This paper presents a texture-specific BoVW method to represent focal liver lesions (FLLs). Pixels in the region of interest (ROI) are classified into nine texture categories using the rotation-invariant uniform local binary pattern method. The BoVW-based features are calculated for each texture category. In addition, a spatial cone matching (SCM)-based representation strategy is proposed to describe the spatial information of the visual words in the ROI. In a pilot study, eight radiologists with different clinical experience performed diagnoses for 20 cases with and without the top six retrieved results. A total of 132 multiphase computed tomography volumes including five pathological types were collected. The texture-specific BoVW was compared to other BoVW-based methods using the constructed dataset of FLLs. The results show that our proposed model outperforms the other three BoVW methods in discriminating different lesions. The SCM method, which adds spatial information to the orderless BoVW model, impacted the retrieval performance. In the pilot trial, the average diagnosis accuracy of the radiologists was improved from 66 to 80% using the retrieval system. The preliminary results indicate that the texture-specific features and the SCM-based BoVW features can effectively characterize various liver lesions. The retrieval system has the potential to improve the diagnostic accuracy and the confidence of the radiologists.

Preparation and surface characterization of plasma-treated and biomolecular-micropatterned polymer substrates

NASA Astrophysics Data System (ADS)

Langowski, Bryan Alfred

A micropatterning process creates distinct microscale domains on substrate surfaces that differ from the surfaces' original chemical/physical properties. Numerous micropatterning methods exist, each having relative advantages and disadvantages in terms of cost, ease, reproducibility, and versatility. Polymeric surfaces micropatterned with biomolecules have many applications, but are specifically utilized in tissue engineering as cell scaffolds that attempt to controlled tissue generation in vivo and ex vivo. As the physical and chemical cues presented by micropatterned substrates control resulting cellular behavior, characterization of these cues via surface-sensitive analytical techniques is essential in developing cell scaffolds that mimic complex in vivo physicochemical environments. The initial focus of this thesis is the chemical and physical characterization of plasma-treated, microcontact-printed (muCP) polymeric substrates used to direct nerve cell behavior. Unmodified and oxygen plasma-treated poly(methyl methacrylate) (PMMA) substrates were analyzed by surface sensitive techniques to monitor plasma-induced chemical and physical modifications. Additionally, protein-micropattern homogeneity and size were microscopically evaluated. Lastly, poly(dimethylsiloxane) (PDMS) stamps and contaminated PMMA substrates were characterized by spectroscopic and microscopic methods to identify a contamination source during microcontact printing. The final focus of this thesis is the development of microscale plasma-initiated patterning (muPIP) as a versatile, reproducible micropatterning method. Using muPIP, polymeric substrates were micropatterned with several biologically relevant inks. Polymeric substrates were characterized following muPIP by surface-sensitive techniques to identify the technique's underlying physical and chemical bases. In addition, neural stem cell response to muPIP-generated laminin micropatterns was microscopically and biologically evaluated. Finally, enhanced versatility of muPIP in generating microscale poly-L-lysine gradients was demonstrated.

Magnetic Resonance Imaging of Ventilation and Perfusion in the Lung

NASA Technical Reports Server (NTRS)

Prisk, Gordon Kim (Inventor); Hopkins, Susan Roberta (Inventor); Pereira De Sa, Rui Carlos (Inventor); Theilmann, Rebecca Jean (Inventor); Buxton, Richard Bruce (Inventor); Cronin, Matthew Vincent (Inventor)

2017-01-01

Methods, devices, and systems are disclosed for implementing a fully quantitative non-injectable contrast proton MRI technique to measure spatial ventilation-perfusion (VA/Q) matching and spatial distribution of ventilation and perfusion. In one aspect, a method using MRI to characterize ventilation and perfusion in a lung includes acquiring an MR image of the lung having MR data in a voxel and obtaining a breathing frequency parameter, determining a water density value, a specific ventilation value, and a perfusion value in at least one voxel of the MR image based on the MR data and using the water density value to determine an air content value, and determining a ventilation-perfusion ratio value that is the product of the specific ventilation value, the air content value, the inverse of the perfusion value, and the breathing frequency.

Characterization of Interfacial Chemistry of Adhesive/Dentin Bond Using FTIR Chemical Imaging With Univariate and Multivariate Data Processing

PubMed Central

Wang, Yong; Yao, Xiaomei; Parthasarathy, Ranganathan

2008-01-01

Fourier transform infrared (FTIR) chemical imaging can be used to investigate molecular chemical features of the adhesive/dentin interfaces. However, the information is not straightforward, and is not easily extracted. The objective of this study was to use multivariate analysis methods, principal component analysis and fuzzy c-means clustering, to analyze spectral data in comparison with univariate analysis. The spectral imaging data collected from both the adhesive/healthy dentin and adhesive/caries-affected dentin specimens were used and compared. The univariate statistical methods such as mapping of intensities of specific functional group do not always accurately identify functional group locations and concentrations due to more or less band overlapping in adhesive and dentin. Apart from the ease with which information can be extracted, multivariate methods highlight subtle and often important changes in the spectra that are difficult to observe using univariate methods. The results showed that the multivariate methods gave more satisfactory, interpretable results than univariate methods and were conclusive in showing that they can discriminate and classify differences between healthy dentin and caries-affected dentin within the interfacial regions. It is demonstrated that the multivariate FTIR imaging approaches can be used in the rapid characterization of heterogeneous, complex structure. PMID:18980198

Hierarchical structures of amorphous solids characterized by persistent homology

PubMed Central

Hiraoka, Yasuaki; Nakamura, Takenobu; Hirata, Akihiko; Escolar, Emerson G.; Matsue, Kaname; Nishiura, Yasumasa

2016-01-01

This article proposes a topological method that extracts hierarchical structures of various amorphous solids. The method is based on the persistence diagram (PD), a mathematical tool for capturing shapes of multiscale data. The input to the PDs is given by an atomic configuration and the output is expressed as 2D histograms. Then, specific distributions such as curves and islands in the PDs identify meaningful shape characteristics of the atomic configuration. Although the method can be applied to a wide variety of disordered systems, it is applied here to silica glass, the Lennard-Jones system, and Cu-Zr metallic glass as standard examples of continuous random network and random packing structures. In silica glass, the method classified the atomic rings as short-range and medium-range orders and unveiled hierarchical ring structures among them. These detailed geometric characterizations clarified a real space origin of the first sharp diffraction peak and also indicated that PDs contain information on elastic response. Even in the Lennard-Jones system and Cu-Zr metallic glass, the hierarchical structures in the atomic configurations were derived in a similar way using PDs, although the glass structures and properties substantially differ from silica glass. These results suggest that the PDs provide a unified method that extracts greater depth of geometric information in amorphous solids than conventional methods. PMID:27298351

Minimum information specification for in situ hybridization and immunohistochemistry experiments (MISFISHIE).

PubMed

Deutsch, Eric W; Ball, Catherine A; Berman, Jules J; Bova, G Steven; Brazma, Alvis; Bumgarner, Roger E; Campbell, David; Causton, Helen C; Christiansen, Jeffrey H; Daian, Fabrice; Dauga, Delphine; Davidson, Duncan R; Gimenez, Gregory; Goo, Young Ah; Grimmond, Sean; Henrich, Thorsten; Herrmann, Bernhard G; Johnson, Michael H; Korb, Martin; Mills, Jason C; Oudes, Asa J; Parkinson, Helen E; Pascal, Laura E; Pollet, Nicolas; Quackenbush, John; Ramialison, Mirana; Ringwald, Martin; Salgado, David; Sansone, Susanna-Assunta; Sherlock, Gavin; Stoeckert, Christian J; Swedlow, Jason; Taylor, Ronald C; Walashek, Laura; Warford, Anthony; Wilkinson, David G; Zhou, Yi; Zon, Leonard I; Liu, Alvin Y; True, Lawrence D

2008-03-01

One purpose of the biomedical literature is to report results in sufficient detail that the methods of data collection and analysis can be independently replicated and verified. Here we present reporting guidelines for gene expression localization experiments: the minimum information specification for in situ hybridization and immunohistochemistry experiments (MISFISHIE). MISFISHIE is modeled after the Minimum Information About a Microarray Experiment (MIAME) specification for microarray experiments. Both guidelines define what information should be reported without dictating a format for encoding that information. MISFISHIE describes six types of information to be provided for each experiment: experimental design, biomaterials and treatments, reporters, staining, imaging data and image characterizations. This specification has benefited the consortium within which it was developed and is expected to benefit the wider research community. We welcome feedback from the scientific community to help improve our proposal.

A spatially-explicit data driven approach to assess the effect of agricultural land occupation on species groups

NASA Astrophysics Data System (ADS)

Elshout, P.; van Zelm, R.; Karuppiah, R.; Laurenzi, I.; Huijbregts, M.

2013-12-01

Change of vegetation cover and increased land use intensity can directly affect the natural habitat and the wildlife it houses. The actual impact of agricultural land use is region specific as crops are grown under various climatic conditions and ways of cultivation and refining. Furthermore, growing a specific crop in a tropical region may require clearance of rainforest while the same crop may replace natural grasslands in temperate regions. Within life cycle impact assessment (LCIA), methods to address impacts of land use on a global scale are still in need of development. We aim to extend existing methods to improve the robustness of LCIA by allowing spatial differentiation of agricultural land use impacts. The goal of this study is to develop characterization factors for the direct impact of land use on biodiversity, which results from the replacement of natural habitat with farmland. The characterization factor expresses the change in species richness under crop cultivation compared to the species richness in the natural situation over a certain area. A second goal was to identify the differences in impacts caused by cultivation of different crop types, sensitivity of different taxonomic groups, and differences in natural land cover. Empirical data on species richness were collected from literature for both natural reference situations and agricultural land use situations. Reference situations were selected on an ecoregion or biome basis. We calculated characterization factors for four crop groups (oil palm, low crops, cereals, and perennial grasses), four species groups (arthropods, birds, mammals, vascular plants), and six biomes.

Fast Low-Rank Shared Dictionary Learning for Image Classification.

PubMed

Tiep Huu Vu; Monga, Vishal

2017-11-01

Despite the fact that different objects possess distinct class-specific features, they also usually share common patterns. This observation has been exploited partially in a recently proposed dictionary learning framework by separating the particularity and the commonality (COPAR). Inspired by this, we propose a novel method to explicitly and simultaneously learn a set of common patterns as well as class-specific features for classification with more intuitive constraints. Our dictionary learning framework is hence characterized by both a shared dictionary and particular (class-specific) dictionaries. For the shared dictionary, we enforce a low-rank constraint, i.e., claim that its spanning subspace should have low dimension and the coefficients corresponding to this dictionary should be similar. For the particular dictionaries, we impose on them the well-known constraints stated in the Fisher discrimination dictionary learning (FDDL). Furthermore, we develop new fast and accurate algorithms to solve the subproblems in the learning step, accelerating its convergence. The said algorithms could also be applied to FDDL and its extensions. The efficiencies of these algorithms are theoretically and experimentally verified by comparing their complexities and running time with those of other well-known dictionary learning methods. Experimental results on widely used image data sets establish the advantages of our method over the state-of-the-art dictionary learning methods.

Fast Low-Rank Shared Dictionary Learning for Image Classification

NASA Astrophysics Data System (ADS)

Vu, Tiep Huu; Monga, Vishal

2017-11-01

Despite the fact that different objects possess distinct class-specific features, they also usually share common patterns. This observation has been exploited partially in a recently proposed dictionary learning framework by separating the particularity and the commonality (COPAR). Inspired by this, we propose a novel method to explicitly and simultaneously learn a set of common patterns as well as class-specific features for classification with more intuitive constraints. Our dictionary learning framework is hence characterized by both a shared dictionary and particular (class-specific) dictionaries. For the shared dictionary, we enforce a low-rank constraint, i.e. claim that its spanning subspace should have low dimension and the coefficients corresponding to this dictionary should be similar. For the particular dictionaries, we impose on them the well-known constraints stated in the Fisher discrimination dictionary learning (FDDL). Further, we develop new fast and accurate algorithms to solve the subproblems in the learning step, accelerating its convergence. The said algorithms could also be applied to FDDL and its extensions. The efficiencies of these algorithms are theoretically and experimentally verified by comparing their complexities and running time with those of other well-known dictionary learning methods. Experimental results on widely used image datasets establish the advantages of our method over state-of-the-art dictionary learning methods.

Development of a method for urine bikunin/urinary trypsin inhibitor (UTI) quantitation and structural characterization: Application to type 1 and type 2 diabetes.

PubMed

Lepedda, Antonio Junior; Nieddu, Gabriele; Rocchiccioli, Silvia; Fresu, Pietro; De Muro, Pierina; Formato, Marilena

2013-12-01

Bikunin is a plasma proteinase inhibitor often associated with inflammatory conditions. It has a half-life of few minutes and it is rapidly excreted into urine as urinary trypsin inhibitor (UTI). UTI levels are usually low in healthy individuals but they can increase up to tenfold in both acute and chronic inflammatory diseases. This article describes a sensitive method for both direct UTI quantitation and structural characterization. UTI purification was performed by anion exchange micro-chromatography followed by SDS-PAGE. A calibration curve for protein quantitation was set up by using a purified UTI fraction. UTI identification and structural characterization was performed by Nano-LC-MS/MS analysis. The method was applied on urine samples from 9 patients with type 1 diabetes, 11 patients with type 2 diabetes, and 28 healthy controls, matched for age and sex with patients, evidencing higher UTI levels in both groups of patients with respect to controls (p < 0.001 and p = 0.001, respectively). Spearman's correlation tests highlighted no association between UTI levels and age in each group tested. Owing to the elevated sensitivity and specificity, the described method allows UTI quantitation from very low quantities of specimen. Furthermore, as UTI concentration is normalized for creatinine level, the analysis could be also performed on randomly collected urine samples. Finally, MS/MS analysis prospects the possibility of characterizing PTM sites potentially able to affect UTI localization, function, and pathophysiological activity. Preliminary results suggest that UTI levels could represent a useful marker of chronic inflammatory condition in type 1 and 2 diabetes. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chromatographic test methods for characterizing alkylsiloxane-bonded silica columns for reversed-phase liquid chromatography.

PubMed

Poole, Colin F

2018-06-07

Major obstacles to formulating a simple retention mechanism for reversed-phase liquid chromatography have a direct impact on the development of experimental methods for column characterization as they limit our capability to understand observed differences in retention at a system level. These problems arise from the heterogeneous composition of the stationary phase, the difficulty of providing a working definition for the phase ratio, and uncertainty as to whether the distribution mechanism for varied compounds is a partition, adsorption or mixed (combination) of these models. Retention factor and separation factor measurements offer little guidance as they represent an average of various and variable contributing factors that can only be interpreted by assuming a specific model. Column characterization methods have tended to ignore these difficulties by inventing a series of terms to describe column properties, such as hydrophobicity, hydrophilicity, silanol activity, steric resistance, etc., without proper definition. This has allowed multiple scales to be proposed for the same property which generally are only weakly correlated. Against this background we review the major approaches for the characterization of alkylsiloxane-bonded silica stationary phases employing prototypical compounds, the hydrophobic-subtraction model and the solvation parameter model. Those methods using prototypical compounds are limited by the lack of compounds with a singular dominant interaction. The multivariate approaches that extract column characteristic properties from the retention of varied compounds are more hopeful but it is important to be more precise in defining the characteristic column properties and cognizant that general interpretation of these properties for varied columns cannot escape the problem of a poor understanding of the distribution mechanism. Copyright © 2018 Elsevier B.V. All rights reserved.

Characterization of Contamination Generation Characteristics of Satellite Materials

DTIC Science & Technology

1989-11-22

higher than ambient because of sorption interactions between outgassed species and the chamber walls. In the mass collection method, the mate ’al sample...data run being acquired or the processing of previously acquired data. The Wiley/JNBS standard mass spectra library containing over 70,000 compounds ...outgassing for the specific test material. Outgasslng from materials such as adhesives and potting compounds is diffusion- controlled, so the outgaving rate

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

Couture, John J.; Singh, Aditya; Rubert-Nason, Kennedy F.

Spectroscopy has recently emerged as an effective method to accurately characterize leaf biochemistry in living tissue through the application of chemometric approaches to foliar optical data, but this approach has not been widely used for plant secondary metabolites. Here in this paper, we examine the ability of reflectance spectroscopy to quantify specific phenolic compounds in trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera) that play influential roles in ecosystem functioning related to trophic-level interactions and nutrient cycling.

Work plan for the identification of techniques for in-situ sensing of layering/interfaces of Hanford high level waste tank

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

Vargo, G.F. Jr.

1995-06-16

The purpose of this work scope is to identify a specific potential technology/device/instrument/ideas that would provide the tank waste data. A method is needed for identifying layering and physical state within the large waste tanks at the Hanford site in Washington State. These interfaces and state changes can adversely impact sampling and characterization activities.

Sintered rare earth-iron Laves phase magnetostrictive alloy product and preparation thereof

DOEpatents

Malekzadeh, Manoochehr; Pickus, Milton R.

1979-01-01

A sintered rare earth-iron Laves phase magnetostrictive alloy product characterized by a grain oriented morphology. The grain oriented morphology is obtained by magnetically aligning powder particles of the magnetostrictive alloy prior to sintering. Specifically disclosed are grain oriented sintered compacts of Tb.sub.x Dy.sub.1-x Fe.sub.2 and their method of preparation. The present sintered products have enhanced magnetostrictive properties.

Development, Application, and Implementation of RAMCAP to Characterize Nuclear Power Plant Risk From Terrorism

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

Gaertner, John P.; Teagarden, Grant A.

2006-07-01

In response to increased interest in risk-informed decision making regarding terrorism, EPRI and ERIN Engineering were selected by U.S. DHS and ASME to develop and demonstrate the RAMCAP method for nuclear power plant (NPP) risk assessment. The objective is to characterize plant-specific NPP risk for risk management opportunities and to provide consistent information for DHS decision making. This paper is an update of this project presented at the American Nuclear Society (ANS) International Topical Meeting on Probabilistic Safety Analysis (PSA05) in September, 2005. The method uses a characterization of risk as a function of Consequence, Vulnerability, and Threat. For eachmore » site, worst case scenarios are developed for each of sixteen benchmark threats. Nuclear RAMCAP hypothesizes that the intent of the perpetrator is to cause offsite radiological consequences. Specific targets are the reactor core, the spent fuel pool, and nuclear spent fuel in a dry storage facility (ISFSI). Results for each scenario are presented as conditional risk for financial loss, early fatalities and early injuries. Expected consequences for each scenario are quantified, while vulnerability is estimated on a relative likelihood scale. Insights for other societal risks are provided. Although threat frequencies are not provided, target attractiveness and threat deterrence are estimated. To assure efficiency, completeness, and consistency; results are documented using standard RAMCAP Evaluator software. Trial applications were successfully performed at four plant sites. Implementation at all other U.S. commercial sites is underway, supported by the Nuclear Sector Coordinating Council (NSCC). Insights from RAMCAP results at 23 U.S. plants completed to date have been compiled and presented to the NSCC. Results are site-specific. Physical security barriers, an armed security force, preparedness for design-basis threats, rugged design against natural hazards, multiple barriers between fuel and environment, accident mitigation capability, severe accident management procedures, and offsite emergency plans are risk-beneficial against all threat types. (authors)« less

Unsupervised machine-learning method for improving the performance of ambulatory fall-detection systems

PubMed Central

2012-01-01

Background Falls can cause trauma, disability and death among older people. Ambulatory accelerometer devices are currently capable of detecting falls in a controlled environment. However, research suggests that most current approaches can tend to have insufficient sensitivity and specificity in non-laboratory environments, in part because impacts can be experienced as part of ordinary daily living activities. Method We used a waist-worn wireless tri-axial accelerometer combined with digital signal processing, clustering and neural network classifiers. The method includes the application of Discrete Wavelet Transform, Regrouping Particle Swarm Optimization, Gaussian Distribution of Clustered Knowledge and an ensemble of classifiers including a multilayer perceptron and Augmented Radial Basis Function (ARBF) neural networks. Results Preliminary testing with 8 healthy individuals in a home environment yields 98.6% sensitivity to falls and 99.6% specificity for routine Activities of Daily Living (ADL) data. Single ARB and MLP classifiers were compared with a combined classifier. The combined classifier offers the greatest sensitivity, with a slight reduction in specificity for routine ADL and an increased specificity for exercise activities. In preliminary tests, the approach achieves 100% sensitivity on in-group falls, 97.65% on out-group falls, 99.33% specificity on routine ADL, and 96.59% specificity on exercise ADL. Conclusion The pre-processing and feature-extraction steps appear to simplify the signal while successfully extracting the essential features that are required to characterize a fall. The results suggest this combination of classifiers can perform better than MLP alone. Preliminary testing suggests these methods may be useful for researchers who are attempting to improve the performance of ambulatory fall-detection systems. PMID:22336100

CRAX/Cassandra Reliability Analysis Software

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

Robinson, D.

1999-02-10

Over the past few years Sandia National Laboratories has been moving toward an increased dependence on model- or physics-based analyses as a means to assess the impact of long-term storage on the nuclear weapons stockpile. These deterministic models have also been used to evaluate replacements for aging systems, often involving commercial off-the-shelf components (COTS). In addition, the models have been used to assess the performance of replacement components manufactured via unique, small-lot production runs. In either case, the limited amount of available test data dictates that the only logical course of action to characterize the reliability of these components ismore » to specifically consider the uncertainties in material properties, operating environment etc. within the physics-based (deterministic) model. This not only provides the ability to statistically characterize the expected performance of the component or system, but also provides direction regarding the benefits of additional testing on specific components within the system. An effort was therefore initiated to evaluate the capabilities of existing probabilistic methods and, if required, to develop new analysis methods to support the inclusion of uncertainty in the classical design tools used by analysts and design engineers at Sandia. The primary result of this effort is the CMX (Cassandra Exoskeleton) reliability analysis software.« less

Artificial intelligence techniques for colorectal cancer drug metabolism: ontology and complex network.

PubMed

Martínez-Romero, Marcos; Vázquez-Naya, José M; Rabuñal, Juan R; Pita-Fernández, Salvador; Macenlle, Ramiro; Castro-Alvariño, Javier; López-Roses, Leopoldo; Ulla, José L; Martínez-Calvo, Antonio V; Vázquez, Santiago; Pereira, Javier; Porto-Pazos, Ana B; Dorado, Julián; Pazos, Alejandro; Munteanu, Cristian R

2010-05-01

Colorectal cancer is one of the most frequent types of cancer in the world and generates important social impact. The understanding of the specific metabolism of this disease and the transformations of the specific drugs will allow finding effective prevention, diagnosis and treatment of the colorectal cancer. All the terms that describe the drug metabolism contribute to the construction of ontology in order to help scientists to link the correlated information and to find the most useful data about this topic. The molecular components involved in this metabolism are included in complex network such as metabolic pathways in order to describe all the molecular interactions in the colorectal cancer. The graphical method of processing biological information such as graphs and complex networks leads to the numerical characterization of the colorectal cancer drug metabolic network by using invariant values named topological indices. Thus, this method can help scientists to study the most important elements in the metabolic pathways and the dynamics of the networks during mutations, denaturation or evolution for any type of disease. This review presents the last studies regarding ontology and complex networks of the colorectal cancer drug metabolism and a basic topology characterization of the drug metabolic process sub-ontology from the Gene Ontology.

Characterization of Aluminum Honeycomb and Experimentation for Model Development and Validation, Volume I: Discovery and Characterization Experiments for High-Density Aluminum Honeycomb

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

Lu, Wei-Yang; Korellis, John S.; Lee, Kenneth L.

2006-08-01

Honeycomb is a structure that consists of two-dimensional regular arrays of open cells. High-density aluminum honeycomb has been used in weapon assemblies to mitigate shock and protect payload because of its excellent crush properties. In order to use honeycomb efficiently and to certify the payload is protected by the honeycomb under various loading conditions, a validated honeycomb crush model is required and the mechanical properties of the honeycombs need to be fully characterized. Volume I of this report documents an experimental study of the crush behavior of high-density honeycombs. Two sets of honeycombs were included in this investigation: commercial grademore » for initial exploratory experiments, and weapon grade, which satisfied B61 specifications. This investigation also includes developing proper experimental methods for crush characterization, conducting discovery experiments to explore crush behaviors for model improvement, and identifying experimental and material uncertainties.« less

Development of characterization protocol for mixed liquid radioactive waste classification

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

Zakaria, Norasalwa, E-mail: norasalwa@nuclearmalaysia.gov.my; Wafa, Syed Asraf; Wo, Yii Mei

2015-04-29

Mixed liquid organic waste generated from health-care and research activities containing tritium, carbon-14, and other radionuclides posed specific challenges in its management. Often, these wastes become legacy waste in many nuclear facilities and being considered as ‘problematic’ waste. One of the most important recommendations made by IAEA is to perform multistage processes aiming at declassification of the waste. At this moment, approximately 3000 bottles of mixed liquid waste, with estimated volume of 6000 litres are currently stored at the National Radioactive Waste Management Centre, Malaysia and some have been stored for more than 25 years. The aim of this studymore » is to develop a characterization protocol towards reclassification of these wastes. The characterization protocol entails waste identification, waste screening and segregation, and analytical radionuclides profiling using various analytical procedures including gross alpha/ gross beta, gamma spectrometry, and LSC method. The results obtained from the characterization protocol are used to establish criteria for speedy classification of the waste.« less

Procedure de caracterisation thermophysique d'un materiau a changement de phase composite pour le stockage thermique

NASA Astrophysics Data System (ADS)

Le Du, Mathieu

The use of phase change materials (PCMs) allows to store and release large amounts of energy in reduced volumes by using latent heat storage through melting and solidifying at specific temperatures. Phase change materials received a great interest for reducing energy consumption by easing the implementation of passive solar heating and cooling. They can be integrated to buildings as wallboards to improve the heat storage capacity. In this study, an original experimental device has allowed to characterize the thermophysical proprieties of a composite wallboard constituted of PCMs. Generally, PCMs are characterized by calorimetric methods which use very small quantities of material. The device used can characterize large sample's dimensions, as they could be used in real condition. Apparent thermal conductivity and specific heat have been measured for various temperatures. During phase change process, total and latent heat storage capacities have been evaluated with the peak melting and freezing temperatures. Results are compared to the manufacturer's data and data from literature. Incoherencies have been found between sources. Despite several differences with published data, overall results are similar to the latest information, which allow validate the original experimental device. Thermal disturbances due to hysteresis have been noticed and discussed. Results allow suggesting recommendations on thermal procedure and experimental device to characterize efficiently this kind of materials. Temperature's ranges and heating and freezing rates affect results and it must be considered in the characterization. Moreover, experimental devices have to be designed to allow similar heating and freezing rates in order to compare results during melting and freezing. Key words: Phase change material, latent thermal storage, thermophysical characterization.

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.

Characterization of phenol and cresol biodegradation by compound-specific stable isotope analysis.

PubMed

Wei, Xi; Gilevska, Tetyana; Wetzig, Felix; Dorer, Conrad; Richnow, Hans-Hermann; Vogt, Carsten

2016-03-01

Microbial degradation of phenol and cresols can occur under oxic and anoxic conditions by different degradation pathways. One recent technique to take insight into reaction mechanisms is compound-specific isotope analysis (CSIA). While enzymes and reaction mechanisms of several degradation pathways have been characterized in (bio)chemical studies, associated isotope fractionation patterns have been rarely reported, possibly due to constraints in current analytical methods. In this study, carbon enrichment factors and apparent kinetic isotope effects (AKIEc) of the initial steps of different aerobic and anaerobic phenol and cresols degradation pathways were analyzed by isotope ratio mass spectrometry connected with liquid chromatography (LC-IRMS). Significant isotope fractionation was detected for aerobic ring hydroxylation, anoxic side chain hydroxylation, and anoxic fumarate addition, while anoxic carboxylation reactions produced small and inconsistent fractionation. The results suggest that several microbial degradation pathways of phenol and cresols are detectable in the environment by CSIA. Copyright © 2015 Elsevier Ltd. All rights reserved.

Synthesis and characterization of SBA-3, SBA-15, and SBA-1 nanostructured catalytic materials.

PubMed

Anunziata, Oscar A; Beltramone, Andrea R; Martínez, Maria L; Belon, Lizandra López

2007-11-01

A highly ordered large pore mesoporous silica molecular sieve SBA-3, SBA-15, Al-SBA-15, and SBA-1, were developed and characterized by XRD, BET, FTIR, SEM, and NMR-MAS. The catalytic materials were synthesized using different raw materials and operation conditions. These materials contain a regular arrangement of uniform channels with diameters between 1.8 and 10 nm, high specific surface area and high specific pore volume. The designed methods were effective for the synthesis, presenting each mesostructured materials, patterns of XRD and other characteristics corresponding to the reported ones in literature. The new route employed to synthesize Al-SBA-15, generates a catalyst with only aluminum in tetrahedral form, according to the data of (27)Al NMR-MAS. However, several reports indicated that the coordination of the Al atoms changes below the Si/Al ratio of 45, presenting peaks corresponding to penta and hexa-coordinated aluminum, which are absent in our samples (Si/Al = 50 and 33).

Beyond Vmax and Km: How details of enzyme function influence geochemical cycles

NASA Astrophysics Data System (ADS)

Steen, A. D.

2015-12-01

Enzymes catalyze the vast majority of chemical reactions relevant to geomicrobiology. Studies of the activities of enzymes in environmental systems often report Vmax (the maximum possible rate of reaction; often proportional to the concentration of enzymes in the system) and sometimes Km (a measure of the affinity between enzymes and their substrates). However, enzyme studies - particularly those related to enzymes involved in organic carbon oxidation - are often limited to only those parameters, and a relatively limited and mixed set of enzymes. Here I will discuss some novel methods to assay and characterize the specific sets of enzymes that may be important to the carbon cycle in aquatic environments. First, kinetic experiments revealed the collective properties of the complex mixtures of extracellular peptidases that occur where microbial communities are diverse. Crystal structures combined with biochemical characterization of specific enzymes can yield more detailed information about key steps in organic carbon transformations. These new techniques have the potential to provide mechanistic grounding to geomicrobiological models.

Micro-tattoo guided OCT imaging of site specific inflammation

NASA Astrophysics Data System (ADS)

Phillips, Kevin G.; Choudhury, Niloy; Samatham, Ravikant V.; Singh, Harvinder; Jacques, Steven L.

2010-02-01

Epithelial biologists studying human skin diseases such as cancer formation and psoriasis commonly utilize mouse models to characterize the interplay among cells and intracellular signal transduction pathways that result in programmed changes in gene expression and cellular behaviors. The information obtained from animal models is useful only when phenotypic presentations of disease recapitulate those observed in humans. Excision of tissues followed by histochemical analysis is currently the primary means of establishing the morphological presentation. Non invasive imaging of animal models provides an alternate means to characterize tissue morphology associated with the disease of interest in vivo. While useful, the ability to perform in vivo imaging at different time points in the same tissue location has been a challenge. This information is key to understanding site specific changes as the imaged tissue can now be extracted and analyzed for mRNA expression. We present a method employing a micro-tattoo to guide optical coherence tomography (OCT) imaging of ultraviolet induced inflammation over time in the same tissue locations.

Prospecting Metagenomic Enzyme Subfamily Genes for DNA Family Shuffling by a Novel PCR-based Approach*

PubMed Central

Wang, Qiuyan; Wu, Huili; Wang, Anming; Du, Pengfei; Pei, Xiaolin; Li, Haifeng; Yin, Xiaopu; Huang, Lifeng; Xiong, Xiaolong

2010-01-01

DNA family shuffling is a powerful method for enzyme engineering, which utilizes recombination of naturally occurring functional diversity to accelerate laboratory-directed evolution. However, the use of this technique has been hindered by the scarcity of family genes with the required level of sequence identity in the genome database. We describe here a strategy for collecting metagenomic homologous genes for DNA shuffling from environmental samples by truncated metagenomic gene-specific PCR (TMGS-PCR). Using identified metagenomic gene-specific primers, twenty-three 921-bp truncated lipase gene fragments, which shared 64–99% identity with each other and formed a distinct subfamily of lipases, were retrieved from 60 metagenomic samples. These lipase genes were shuffled, and selected active clones were characterized. The chimeric clones show extensive functional and genetic diversity, as demonstrated by functional characterization and sequence analysis. Our results indicate that homologous sequences of genes captured by TMGS-PCR can be used as suitable genetic material for DNA family shuffling with broad applications in enzyme engineering. PMID:20962349

3D print of polymer bonded rare-earth magnets, and 3D magnetic field scanning with an end-user 3D printer

NASA Astrophysics Data System (ADS)

Huber, C.; Abert, C.; Bruckner, F.; Groenefeld, M.; Muthsam, O.; Schuschnigg, S.; Sirak, K.; Thanhoffer, R.; Teliban, I.; Vogler, C.; Windl, R.; Suess, D.

2016-10-01

3D print is a recently developed technique, for single-unit production, and for structures that have been impossible to build previously. The current work presents a method to 3D print polymer bonded isotropic hard magnets with a low-cost, end-user 3D printer. Commercially available isotropic NdFeB powder inside a PA11 matrix is characterized, and prepared for the printing process. An example of a printed magnet with a complex shape that was designed to generate a specific stray field is presented, and compared with finite element simulation solving the macroscopic Maxwell equations. For magnetic characterization, and comparing 3D printed structures with injection molded parts, hysteresis measurements are performed. To measure the stray field outside the magnet, the printer is upgraded to a 3D magnetic flux density measurement system. To skip an elaborate adjusting of the sensor, a simulation is used to calibrate the angles, sensitivity, and the offset of the sensor. With this setup, a measurement resolution of 0.05 mm along the z-axes is achievable. The effectiveness of our calibration method is shown. With our setup, we are able to print polymer bonded magnetic systems with the freedom of having a specific complex shape with locally tailored magnetic properties. The 3D scanning setup is easy to mount, and with our calibration method we are able to get accurate measuring results of the stray field.

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.

Development of SCAR (sequence-characterized amplified region) markers as a complementary tool for identification of ginger (Zingiber officinale Roscoe) from crude drugs and multicomponent formulations.

PubMed

Chavan, Preeti; Warude, Dnyaneshwar; Joshi, Kalpana; Patwardhan, Bhushan

2008-05-01

Zingiber officinale Roscoe (common or culinary ginger) is an official drug in Ayurvedic, Indian herbal, Chinese, Japanese, African and British Pharmacopoeias. The objective of the present study was to develop DNA-based markers that can be applied for the identification and differentiation of the commercially important plant Z. officinale Roscoe from the closely related species Zingiber zerumbet (pinecone, bitter or 'shampoo' ginger) and Zingiber cassumunar [cassumunar or plai (Thai) ginger]. The rhizomes of the other two Zingiber species used in the present study are morphologically similar to that of Z. officinale Roscoe and can be used as its adulterants or contaminants. Various methods, including macroscopy, microscopy and chemoprofiling, have been reported for the quality control of crude ginger and its products. These methods are reported to have limitations in distinguishing Z. officinale from closely related species. Hence, newer complementary methods for correct identification of ginger are useful. In the present study, RAPD (random amplification of polymorphic DNA) analysis was used to identify putative species-specific amplicons for Z. officinale. These were further cloned and sequenced to develop SCAR (sequence-characterized amplified region) markers. The developed SCAR markers were tested in several non-Zingiber species commonly used in ginger-containing formulations. One of the markers, P3, was found to be specific for Z. officinale and was successfully applied for detection of Z. officinale from Trikatu, a multicomponent formulation.

A fractured rock geophysical toolbox method selection tool

USGS Publications Warehouse

Day-Lewis, F. D.; Johnson, C.D.; Slater, L.D.; Robinson, J.L.; Williams, J.H.; Boyden, C.L.; Werkema, D.D.; Lane, J.W.

2016-01-01

Geophysical technologies have the potential to improve site characterization and monitoring in fractured rock, but the appropriate and effective application of geophysics at a particular site strongly depends on project goals (e.g., identifying discrete fractures) and site characteristics (e.g., lithology). No method works at every site or for every goal. New approaches are needed to identify a set of geophysical methods appropriate to specific project goals and site conditions while considering budget constraints. To this end, we present the Excel-based Fractured-Rock Geophysical Toolbox Method Selection Tool (FRGT-MST). We envision the FRGT-MST (1) equipping remediation professionals with a tool to understand what is likely to be realistic and cost-effective when contracting geophysical services, and (2) reducing applications of geophysics with unrealistic objectives or where methods are likely to fail.

Normalization, bias correction, and peak calling for ChIP-seq

PubMed Central

Diaz, Aaron; Park, Kiyoub; Lim, Daniel A.; Song, Jun S.

2012-01-01

Next-generation sequencing is rapidly transforming our ability to profile the transcriptional, genetic, and epigenetic states of a cell. In particular, sequencing DNA from the immunoprecipitation of protein-DNA complexes (ChIP-seq) and methylated DNA (MeDIP-seq) can reveal the locations of protein binding sites and epigenetic modifications. These approaches contain numerous biases which may significantly influence the interpretation of the resulting data. Rigorous computational methods for detecting and removing such biases are still lacking. Also, multi-sample normalization still remains an important open problem. This theoretical paper systematically characterizes the biases and properties of ChIP-seq data by comparing 62 separate publicly available datasets, using rigorous statistical models and signal processing techniques. Statistical methods for separating ChIP-seq signal from background noise, as well as correcting enrichment test statistics for sequence-dependent and sonication biases, are presented. Our method effectively separates reads into signal and background components prior to normalization, improving the signal-to-noise ratio. Moreover, most peak callers currently use a generic null model which suffers from low specificity at the sensitivity level requisite for detecting subtle, but true, ChIP enrichment. The proposed method of determining a cell type-specific null model, which accounts for cell type-specific biases, is shown to be capable of achieving a lower false discovery rate at a given significance threshold than current methods. PMID:22499706

How Analysis Informs Regulation:Success and Failure of ...

EPA Pesticide Factsheets

How Analysis Informs Regulation:Success and Failure of Evolving Approaches to Polyfluoroalkyl Acid Contamination The National Exposure Research Laboratory (NERL) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA mission to protect human health and the environment. HEASD research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.

Developing Decontamination Tools and Approaches to ...

EPA Pesticide Factsheets

Developing Decontamination Tools and Approaches to Address Indoor Pesticide Contamination from Improper Bed Bug Treatments The National Exposure Research Laboratory (NERL) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA mission to protect human health and the environment. HEASD research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.

Advances for the Topographic Characterisation of SMC Materials

PubMed Central

Calvimontes, Alfredo; Grundke, Karina; Müller, Anett; Stamm, Manfred

2009-01-01

For a comprehensive study of Sheet Moulding Compound (SMC) surfaces, topographical data obtained by a contact-free optical method (chromatic aberration confocal imaging) were systematically acquired to characterise these surfaces with regard to their statistical, functional and volumetrical properties. Optimal sampling conditions (cut-off length and resolution) were obtained by a topographical-statistical procedure proposed in the present work. By using different length scales specific morphologies due to the influence of moulding conditions, metallic mould topography, glass fibre content and glass fibre orientation can be characterized. The aim of this study is to suggest a systematic topographical characterization procedure for composite materials in order to study and recognize the influence of production conditions on their surface quality.

Characterization and Discrimination of Gram-Positive Bacteria Using Raman Spectroscopy with the Aid of Principal Component Analysis.

PubMed

Colniță, Alia; Dina, Nicoleta Elena; Leopold, Nicolae; Vodnar, Dan Cristian; Bogdan, Diana; Porav, Sebastian Alin; David, Leontin

2017-09-01

Raman scattering and its particular effect, surface-enhanced Raman scattering (SERS), are whole-organism fingerprinting spectroscopic techniques that gain more and more popularity in bacterial detection. In this work, two relevant Gram-positive bacteria species, Lactobacillus casei ( L. casei ) and Listeria monocytogenes ( L. monocytogenes ) were characterized based on their Raman and SERS spectral fingerprints. The SERS spectra were used to identify the biochemical structures of the bacterial cell wall. Two synthesis methods of the SERS-active nanomaterials were used and the recorded spectra were analyzed. L. casei and L. monocytogenes were successfully discriminated by applying Principal Component Analysis (PCA) to their specific spectral data.

Characterization and Discrimination of Gram-Positive Bacteria Using Raman Spectroscopy with the Aid of Principal Component Analysis

PubMed Central

Leopold, Nicolae; Vodnar, Dan Cristian; Bogdan, Diana; Porav, Sebastian Alin; David, Leontin

2017-01-01

Raman scattering and its particular effect, surface-enhanced Raman scattering (SERS), are whole-organism fingerprinting spectroscopic techniques that gain more and more popularity in bacterial detection. In this work, two relevant Gram-positive bacteria species, Lactobacillus casei (L. casei) and Listeria monocytogenes (L. monocytogenes) were characterized based on their Raman and SERS spectral fingerprints. The SERS spectra were used to identify the biochemical structures of the bacterial cell wall. Two synthesis methods of the SERS-active nanomaterials were used and the recorded spectra were analyzed. L. casei and L. monocytogenes were successfully discriminated by applying Principal Component Analysis (PCA) to their specific spectral data. PMID:28862655

Microinterferometric optical phase tomography for measuring small, asymmetric refractive-index differences in the profiles of optical fibers and fiber devices.

PubMed

Bachim, Brent L; Gaylord, Thomas K

2005-01-20

A new technique, microinterferometric optical phase tomography, is introduced for use in measuring small, asymmetric refractive-index differences in the profiles of optical fibers and fiber devices. The method combines microscopy-based fringe-field interferometry with parallel projection-based computed tomography to characterize fiber index profiles. The theory relating interference measurements to the projection set required for tomographic reconstruction is given, and discrete numerical simulations are presented for three test index profiles that establish the technique's ability to characterize fiber with small, asymmetric index differences. An experimental measurement configuration and specific interferometry and tomography practices employed in the technique are discussed.

Treatability and characterization of Natural Organic Matter (NOM) in South African waters using newly developed methods

NASA Astrophysics Data System (ADS)

Nkambule, T. I.; Krause, R. W. M.; Haarhoff, J.; Mamba, B. B.

Managing the removal of Natural Organic Matter (NOM) or problematic components from water has become increasingly important. NOM is a heterogeneous mixture of organic compounds of human origin and derived from plant and microbial residues. The inadequate removal of NOM has a bearing on the capacity of the other treatment processes to remove organic micro-pollutants or inorganic species that may be present in the water. In addition the action of certain disinfection processes has been shown to lead to the formation of harmful disinfection by-products (DBPs). Owing to the complexity, in composition and structure, of NOM, the techniques currently employed for its characterization have a number of limitations, both in terms of quantification and removal of the NOM within short periods of time. The dissolved organic carbon (DOC), biodegradable dissolved organic carbon (BDOC) and Fluorescence Emission Excitation Matrices (FEEM) were used to characterize NOM from various water samples collected around South Africa. Characterization results gave an indication of the character of NOM present in all the water samples. FEEM and UV-Vis results indicated that most of the water samples were aromatic in nature, since they had high hydrophobic and humic acid-like materials content. Generally, the characterization data indicated a varying composition of NOM amongst the various sampling points. The polarity rapid assessment method (PRAM) was then employed as a rapid NOM characterization tool. The characterization under PRAM is based on preferential adsorption of dissolved organic matter (DOM) fractions onto solid phase extraction (SPE) sorbents. The PRAM also allows the separation of DOM into fractions by polarity, hence reducing the molecular heterogeneity of NOM and thus aiding the removal of specific NOM fractions from water. The PRAM provided a quick characterization of the NOM character. However, DOC quantification by the PRAM analysis was hindered by excessive carbon leaching from the SPE cartridges. The BDOC method of analysis is based on the bacteria fixed on the biologically active sand and gives a ratio of the biodegradable NOM versus the non-biodegradable NOM. For the BDOC analysis, the percentage DOC removal for the samples ranged from 12% to 61%.

Generation of a high-fidelity antibody against nerve growth factor using library scanning mutagenesis and validation with structures of the initial and optimized Fab-antigen complexes

PubMed Central

La Porte, Sherry L; Eigenbrot, Charles; Ultsch, Mark; Ho, Wei-Hsien; Foletti, Davide; Forgie, Alison; Lindquist, Kevin C; Shelton, David L; Pons, Jaume

2014-01-01

Nerve growth factor (NGF) is indispensable during normal embryonic development and critical for the amplification of pain signals in adults. Intervention in NGF signaling holds promise for the alleviation of pain resulting from human diseases such as osteoarthritis, cancer and chronic lower back disorders. We developed a fast, high-fidelity method to convert a hybridoma-derived NGF-targeted mouse antibody into a clinical candidate. This method, termed Library Scanning Mutagenesis (LSM), resulted in the ultra-high affinity antibody tanezumab, a first-in-class anti-hyperalgesic specific for an NGF epitope. Functional and structural comparisons between tanezumab and the mouse 911 precursor antibody using neurotrophin-specific cell survival assays and X-ray crystal structures of both Fab-antigen complexes illustrated high fidelity retention of the NGF epitope. These results suggest the potential for wide applicability of the LSM method for optimization of well-characterized antibodies during humanization. PMID:24830649

Effect of surfactant assisted sonic pretreatment on liquefaction of fruits and vegetable residue: Characterization, acidogenesis, biomethane yield and energy ratio.

PubMed

Shanthi, M; Rajesh Banu, J; Sivashanmugam, P

2018-05-15

The present study explored the disintegration potential of fruits and vegetable residue through sodium dodecyl sulphate (SDS) assisted sonic pretreatment (SSP). In SSP method, initially the biomass barrier (lignin) was removed using SDS at different dosage, subsequently it was sonically disintegrated. The effect of SSP were assessed based on dissolved organic release (DOR) of fruits and vegetable waste and specific energy input. SSP method achieved higher DOR rate and suspended solids reduction (26% and 16%) at optimum SDS dosage of 0.035 g/g SS with least specific energy input of 5400 kJ/kg TS compared to ultrasonic pretreatment (UP) (16% and 10%). The impact of fermentation and biomethane potential assay revealed highest production of volatile fatty acid and methane yield in SSP (1950 mg/L, 0.6 g/g COD) than UP. The energy ratio obtained was 0.9 for SSP, indicating proposed method is energetically efficient. Copyright © 2018 Elsevier Ltd. All rights reserved.

Application of Surface Analysis Methods to Nanomaterials: Summaryof ISO/TC 201 Technical Report: ISO 14187:2011 -Surface Chemical Analysis- Characterization of Nanomaterials

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

Baer, Donald R.

ISO Technical Report (TR) 14187 provides an introduction to (and examples of) the information that can be obtained about nanostructured materials using surface-analysis tools. In addition, both general issues and challenges associated with characterising nanostructured materials and the specific opportunities and challenges associated with individual analytical methods are identified. As the size of objects or components of materials approaches a few nanometres, the distinctions among 'bulk', 'surface' and 'particle' analysis blur. This Technical Report focuses on issues specifically relevant to surface chemical analysis of nanostructured materials. The report considers a variety of analysis methods but focuses on techniques that aremore » in the domain of ISO/TC 201 including Auger electron spectroscopy, X-ray photoelectron spectroscopy, secondary ion mass spectrometry, and scanning probe microscopy. Measurements of nanoparticle surface properties such as surface potential that are often made in a solution are not discussed.« less

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

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

Zhukhlistova, N. E.; Balaev, V. V.; Lyashenko, A. V.

2012-05-15

Endonucleases (EC 3.1) are enzymes of the hydrolase class that catalyze the hydrolytic cleavage of deoxyribonucleic and ribonucleic acids at any region of the polynucleotide chain. Endonucleases are widely used both in biotechnological processes and in veterinary medicine as antiviral agents. Medical applications of endonucleases in human cancer therapy hold promise. The results of X-ray diffraction studies of the spatial organization of endonucleases and their complexes and the mechanism of their action are analyzed and generalized. An analysis of the structural studies of this class of enzymes showed that the specific binding of enzymes to nucleic acids is characterized bymore » interactions with nitrogen bases and the nucleotide backbone, whereas the nonspecific binding of enzymes is generally characterized by interactions only with the nucleic-acid backbone. It should be taken into account that the specificity can be modulated by metal ions and certain low-molecular-weight organic compounds. To test the hypotheses about specific and nonspecific nucleic-acid-binding proteins, it is necessary to perform additional studies of atomic-resolution three-dimensional structures of enzyme-nucleic-acid complexes by methods of structural biology.« less

Heterogeneous growth-induced prestrain in the heart

PubMed Central

Genet, M.; Rausch, M.; Lee, L.C.; Choy, S.; Zhao, X.; Kassab, G.S.; Kozerke, S.; Guccione, J.M.; Kuhl, E.

2015-01-01

Even when entirely unloaded, biological structures are not stress-free, as shown by Y.C. Fung’s seminal opening angle experiment on arteries and the left ventricle. As a result of this prestrain, subject-specific geometries extracted from medical imaging do not represent an unloaded reference configuration necessary for mechanical analysis, even if the structure is externally unloaded. Here we propose a new computational method to create physiological residual stress fields in subject-specific left ventricular geometries using the continuum theory of fictitious configurations combined with a fixed-point iteration. We also reproduced the opening angle experiment on four swine models, to characterize the range of normal opening angle values. The proposed method generates residual stress fields which can reliably reproduce the range of opening angles between 8.7±1.8 and 16.6 ± 13.7 as measured experimentally. We demonstrate that including the effects of prestrain reduces the left ventricular stiffness by up to 40%, thus facilitating the ventricular filling, which has a significant impact on cardiac function. This method can improve the fidelity of subject-specific models to improve our understanding of cardiac diseases and to optimize treatment options. PMID:25913241

Graphene/semicrystalline-carbon derived from amylose films for supercapacitor application

NASA Astrophysics Data System (ADS)

Deraman, M.; Sazali, N. E. S.; Hanappi, M. F. Y. M.; Tajuddin, N. S. M.; Hamdan, E.; Suleman, M.; Othman, M. A. R.; Omar, R.; Hashim, M. A.; Basri, N. H.; Nor, N. S. M.; Dolah, B. N. M.; Noor, A. M.; Jasni, M. R. M.

2016-08-01

Graphene/semicrystalline-carbon in the form of carbon flakes is produced by carbonization up to 600, 700, 800, 900 and 1000°C, respectively, of the amylose films prepared by a casting method on copper foil substrate. The carbon flakes are characterized by X-ray diffraction (XRD) method to determine their microcrystallite interlayer spacing, width and stack-height; and Raman spectroscopy (RS) method to obtain structural information from the D-, D2- and G-bands peak-intensities. The XRD results show that increase in carbonization temperature lead to ~(1-3%), ~85% and ~30%increase in the microcrystallites interlayer spacing, width and stack-height, respectively, indicating that a larger growth of microcrytallite of carbon flakes occurs in the direction parallel to (001) plane or film planar surface. The specific surface area of carbon flakes estimated from the XRD results in decreases from ~4400 to ~3400 m2/g, corresponding to the specific capacitance between ~500 to ~400 F/g, which are well within the range of specific capacitance for typical electrodes carbon for supercapacitor application. The RS results show that the multilayer graphene co-exist with semicrystalline- carbon within the carbon flakes, with the multilayer graphene relative quantities increase with increasing carbonization temperature.

Real-time specific surface area measurements via laser-induced breakdown spectroscopy

USGS Publications Warehouse

Washburn, Kathryn E.; Birdwell, Justin E.; Howard, James E.

2017-01-01

From healthcare to cosmetics to environmental science, the specific surface area (SSA) of micro- and mesoporous materials or products can greatly affect their chemical and physical properties. SSA results are also widely used to examine source rocks in conventional and unconventional petroleum resource plays. Despite its importance, current methods to measure SSA are often cumbersome, time-consuming, or require cryogenic consumables (e.g., liquid nitrogen). These methods are not amenable to high-throughput environments, have stringent sample preparation requirements, and are not practical for use in the field. We present a new application of laser-induced breakdown spectroscopy for rapid measurement of SSA. This study evaluates geological samples, specifically organic-rich oil shales, but the approach is expected to be applicable to many other types of materials. The method uses optical emission spectroscopy to examine laser-generated plasma and quantify the amount of argon adsorbed to a sample during an inert gas purge. The technique can accommodate a wide range of sample sizes and geometries and has the potential for field use. These advantages for SSA measurement combined with the simultaneous acquisition of composition information make this a promising new approach for characterizing geologic samples and other materials.

Microbial source tracking in shellfish harvesting waters in the Gulf of Nicoya, Costa Rica.

PubMed

Symonds, E M; Young, S; Verbyla, M E; McQuaig-Ulrich, S M; Ross, E; Jiménez, J A; Harwood, V J; Breitbart, M

2017-03-15

Current microbial water quality monitoring is generally limited to culture-based measurements of fecal indicator bacteria (FIB). Given the many possible sources of fecal pollution within a watershed and extra-intestinal FIB reservoirs, it is important to determine source(s) of fecal pollution as a means to improve water quality and protect public health. The principal objective of this investigation was to characterize the microbial water quality of shellfish harvesting areas in the Gulf of Nicoya, Costa Rica during 2015. In order to achieve this objective, the specificity and sensitivity of 11 existing microbial source tracking (MST) PCR assays, associated with cows (BacCow), dogs (BacCan, DogBac), domestic wastewater (PMMoV), general avian (GFD), gulls (Gull2), horses (HorseBac, HoF), humans (HF183, HPyV), and pigs (PF), were evaluated using domestic wastewater and animal fecal samples collected from the region. The sensitivity of animal-associated assays ranged from 13 to 100%, while assay specificity ranged from 38 to 100%. The specificity of pepper mild mottle virus (PMMoV) and human polyomavirus (HPyV) was 100% for domestic wastewater, as compared to 94% specificity of the HF183 Bacteroidales marker. PMMoV was identified as a useful domestic wastewater-associated marker, with concentrations as high as 1.1 × 10 5 copies/ml and 100% sensitivity and specificity. Monthly surface water samples collected from four shellfish harvesting areas were analyzed using culture-based methods for Escherichia coli as well as molecular methods for FIB and a suite of MST markers, which were selected for their specificity in the region. While culturable E. coli results suggested possible fecal pollution during the monitoring period, the absence of human/domestic wastewater-associated markers and low FIB concentrations determined using molecular methods indicated sufficient microbial water quality for shellfish harvesting. This is the first study to our knowledge to test the performance of MST markers in Costa Rica as well as in Central America. Given the lack of wastewater treatment and the presence of secondary sources of FIB, this study highlights the importance of an MST toolbox approach to characterize water quality in tropical regions. Furthermore, it confirms and extends the geographic range of PMMoV as an effective tool for monitoring domestic wastewater pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

Serological Evaluation of Immunity to the Varicella-Zoster Virus Based on a Novel Competitive Enzyme-Linked Immunosorbent Assay

PubMed Central

Liu, Jian; Ye, Xiangzhong; Jia, Jizong; Zhu, Rui; Wang, Lina; Chen, Chunye; Yang, Lianwei; Wang, Yongmei; Wang, Wei; Ye, Jianghui; Li, Yimin; Zhu, Hua; Zhao, Qinjian; Cheng, Tong; Xia, Ningshao

2016-01-01

Varicella-zoster virus (VZV) is a highly contagious agent of varicella and herpes zoster. Varicella can be lethal to immunocompromised patients, babies, HIV patients and other adults with impaired immunity. Serological evaluation of immunity to VZV will help determine which individuals are susceptible and evaluate vaccine effectiveness. A collection of 110 monoclonal antibodies (mAbs) were obtained by immunization of mice with membrane proteins or cell-free virus. The mAbs were well characterized, and a competitive sandwich ELISA (capture mAb: 8H6; labelling mAb: 1B11) was established to determine neutralizing antibodies in human serum with reference to the FAMA test. A total of 920 human sera were evaluated. The competitive sandwich ELISA showed a sensitivity of 95.6%, specificity of 99.77% and coincidence of 97.61% compared with the fluorescent-antibody-to-membrane-antigen (FAMA) test. The capture mAb 8H6 was characterized as a specific mAb for VZV ORF9, a membrane-associated tegument protein that interacts with glycoprotein E (gE), glycoprotein B (gB) and glycoprotein C (gC). The labelling mAb 1B11 was characterized as a complement-dependent neutralizing mAb specific for the immune-dominant epitope located on gE, not on other VZV glycoproteins. The established competitive sandwich ELISA could be used as a rapid and high-throughput method for evaluating immunity to VZV. PMID:26853741

Experimental Method for Characterizing Electrical Steel Sheets in the Normal Direction

PubMed Central

Hihat, Nabil; Lecointe, Jean Philippe; Duchesne, Stephane; Napieralska, Ewa; Belgrand, Thierry

2010-01-01

This paper proposes an experimental method to characterise magnetic laminations in the direction normal to the sheet plane. The principle, which is based on a static excitation to avoid planar eddy currents, is explained and specific test benches are proposed. Measurements of the flux density are made with a sensor moving in and out of an air-gap. A simple analytical model is derived in order to determine the permeability in the normal direction. The experimental results for grain oriented steel sheets are presented and a comparison is provided with values obtained from literature. PMID:22163394

Detection of Carbapenemase Production in a Collection of Enterobacteriaceae with Characterized Resistance Mechanisms from Clinical and Environmental Origins by Use of Both Carba NP and Blue-Carba Tests.

PubMed

García-Fernández, Sergio; Morosini, María-Isabel; Gijón, Desirèe; Beatobe, Lorena; Ruiz-Garbajosa, Patricia; Domínguez, Lucas; Cantón, Rafael; Valverde, Aránzazu

2016-02-01

Rapid-screening methods to confirm the presence of resistance mechanisms in multidrug-resistant bacteria are currently recommended. Carba NP and Blue-Carba tests were evaluated in carbapenemase-producing Enterobacteriaceae from hospital (n = 102) and environmental (n = 57) origins for detecting the different molecular classes among them. Both methods showed to be fast and cost-effective, with high sensitivity (98% to 100%) and specificity (100%), and may be easily introduced in the routine laboratory. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Analytical recursive method to ascertain multisite entanglement in doped quantum spin ladders

NASA Astrophysics Data System (ADS)

Roy, Sudipto Singha; Dhar, Himadri Shekhar; Rakshit, Debraj; SenDe, Aditi; Sen, Ujjwal

2017-08-01

We formulate an analytical recursive method to generate the wave function of doped short-range resonating valence bond (RVB) states as a tool to efficiently estimate multisite entanglement as well as other physical quantities in doped quantum spin ladders. We prove that doped RVB ladder states are always genuine multipartite entangled. Importantly, our results show that within specific doping concentration and model parameter regimes, the doped RVB state essentially characterizes the trends of genuine multiparty entanglement in the exact ground states of the Hubbard model with large on-site interactions, in the limit that yields the t -J Hamiltonian.

Microbubble cloud characterization by nonlinear frequency mixing.

PubMed

Cavaro, M; Payan, C; Moysan, J; Baqué, F

2011-05-01

In the frame of the fourth generation forum, France decided to develop sodium fast nuclear reactors. French Safety Authority requests the associated monitoring of argon gas into sodium. This implies to estimate the void fraction, and a histogram indicating the bubble population. In this context, the present letter studies the possibility of achieving an accurate determination of the histogram with acoustic methods. A nonlinear, two-frequency mixing technique has been implemented, and a specific optical device has been developed in order to validate the experimental results. The acoustically reconstructed histograms are in excellent agreement with those obtained using optical methods.

Quantitative analysis of small molecule-nucleic acid interactions with a biosensor surface and surface plasmon resonance detection.

PubMed

Liu, Yang; Wilson, W David

2010-01-01

Surface plasmon resonance (SPR) technology with biosensor surfaces has become a widely-used tool for the study of nucleic acid interactions without any labeling requirements. The method provides simultaneous kinetic and equilibrium characterization of the interactions of biomolecules as well as small molecule-biopolymer binding. SPR monitors molecular interactions in real time and provides significant advantages over optical or calorimetic methods for systems with strong binding coupled to small spectroscopic signals and/or reaction heats. A detailed and practical guide for nucleic acid interaction analysis using SPR-biosensor methods is presented. Details of the SPR technology and basic fundamentals are described with recommendations on the preparation of the SPR instrument, sensor chips, and samples, as well as extensive information on experimental design, quantitative and qualitative data analysis and presentation. A specific example of the interaction of a minor-groove-binding agent with DNA is evaluated by both kinetic and steady-state SPR methods to illustrate the technique. Since the molecules that bind cooperatively to specific DNA sequences are attractive for many applications, a cooperative small molecule-DNA interaction is also presented.

Gender-specific heart rate dynamics in severe intrauterine growth-restricted fetuses.

PubMed

Gonçalves, Hernâni; Bernardes, João; Ayres-de-Campos, Diogo

2013-06-01

Management of intrauterine growth restriction (IUGR) remains a major issue in perinatology. The objective of this paper was the assessment of gender-specific fetal heart rate (FHR) dynamics as a diagnostic tool in severe IUGR. FHR was analyzed in the antepartum period in 15 severe IUGR fetuses and 18 controls, matched for gestational age, in relation to fetal gender. Linear and entropy methods, such as mean FHR (mFHR), low (LF), high (HF) and movement frequency (MF), approximate, sample and multiscale entropy. Sensitivities and specificities were estimated using Fisher linear discriminant analysis and the leave-one-out method. Overall, IUGR fetuses presented significantly lower mFHR and entropy compared with controls. However, gender-specific analysis showed that significantly lower mFHR was only evident in IUGR males and lower entropy in IUGR females. In addition, lower LF/(MF+HF) was patent in IUGR females compared with controls, but not in males. Rather high sensitivities and specificities were achieved in the detection of the FHR recordings related with IUGR male fetuses, when gender-specific analysis was performed at gestational ages less than 34 weeks. Severe IUGR fetuses present gender-specific linear and entropy FHR changes, compared with controls, characterized by a significantly lower entropy and sympathetic-vagal balance in females than in males. These findings need to be considered in order to achieve better diagnostic results. Copyright © 2013 Elsevier Ltd. All rights reserved.

Does fingerprinting truly represent the diversity of wine yeasts? A case study with interdelta genotyping of Saccharomyces cerevisiae strains.

PubMed

Pfliegler, W P; Sipiczki, M

2016-12-01

Simple and efficient genotyping methods are widely used to assess the diversity of a large number of microbial strains, e.g. wine yeasts isolated from a specific geographical area or a vintage. Such methods are often also the first to be applied, to decrease the number of strains deemed interesting for a more time-consuming physiological characterization. Here, we aimed to use a physiologically characterized strain collection of 69 Saccharomyces cerevisiae strains from Hungarian wine regions to determine whether geographical origin or physiological similarity can be recovered by clustering the strains with one or two simultaneously used variations of interdelta genotyping. Our results indicate that although a detailed clustering with high resolution can be achieved with this method, the clustering of strains is largely contrasting when different primer sets are used and it does not recover geographical or physiological groups. Genotyping is routinely used for assessing the diversity of a large number of isolates/strains of a single species, e.g. a collection of wine yeasts. We tested the efficiency of interdelta genotyping on a collection of Saccharomyces wine yeasts from four wine regions of Hungary that was previously characterized physiologically. Interdelta fingerprinting recovered neither physiological nor geographical similarities, and in addition, the two different primer pairs widely used for this method showed conflicting and barely comparable results. Thus, this method does not necessarily represent the true diversity of a strain collection, but detailed clustering may be achieved by the combined use of primer sets. © 2016 The Society for Applied Microbiology.

ASTER preflight and inflight calibration and the validation of level 2 products

USGS Publications Warehouse

Thome, K.; Aral, K.; Hook, S.; Kieffer, H.; Lang, H.; Matsunaga, T.; Ono, A.; Palluconi, F. D.; Sakuma, H.; Slater, P.; Takashima, T.; Tonooka, H.; Tsuchida, S.; Welch, R.M.; Zalewski, E.

1998-01-01

This paper describes the preflight and inflight calibration approaches used for the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER). The system is a multispectral, high-spatial resolution sensor on the Earth Observing System's (EOS)-AMl platform. Preflight calibration of ASTER uses well-characterized sources to provide calibration and preflight round-robin exercises to understand biases between the calibration sources of ASTER and other EOS sensors. These round-robins rely on well-characterized, ultra-stable radiometers. An experiment held in Yokohama, Japan, showed that the output from the source used for the visible and near-infrared (VNIR) subsystem of ASTER may be underestimated by 1.5%, but this is still within the 4% specification for the absolute, radiometric calibration of these bands. Inflight calibration will rely on vicarious techniques and onboard blackbodies and lamps. Vicarious techniques include ground-reference methods using desert and water sites. A recent joint field campaign gives confidence that these methods currently provide absolute calibration to better than 5%, and indications are that uncertainties less than the required 4% should be achievable at launch. The EOS-AMI platform will also provide a spacecraft maneuver that will allow ASTER to see the moon, allowing further characterization of the sensor. A method for combining the results of these independent calibration results is presented. The paper also describes the plans for validating the Level 2 data products from ASTER. These plans rely heavily upon field campaigns using methods similar to those used for the ground-reference, vicarious calibration methods. ?? 1998 IEEE.

Group theoretical derivation of the minimal coupling principle

NASA Astrophysics Data System (ADS)

Nisticò, Giuseppe

2017-04-01

The group theoretical methods worked out by Bargmann, Mackey and Wigner, which deductively establish the Quantum Theory of a free particle for which Galileian transformations form a symmetry group, are extended to the case of an interacting particle. In doing so, the obstacles caused by loss of symmetry are overcome. In this approach, specific forms of the wave equation of an interacting particle, including the equation derived from the minimal coupling principle, are implied by particular first-order invariance properties that characterize the interaction with respect to specific subgroups of Galileian transformations; moreover, the possibility of yet unknown forms of the wave equation is left open.

[Utilization of nylon membranes for specific isolation and characterization of verotoxin-producing Escherichia coli using DNA probes].

PubMed

Gallien, P; Klie, H; Perlberg, K W; Protz, D

1996-01-01

A method for specific isolation of VT(+)-strains in raw milk is given. DNA-hybridization technique with DIG-labeled PCR-amplificates as probes are the basis. No background is seen by using "DIG Easy Hyb" solution and nylon membranes for colony- and plaque-hybridization (Boehringer Mannheim GmbH). Marked colonies are visible on the membranes after detection. So it is possible to select these colonies from a masterplate. The results are available within one day (without enrichment and membrane preparation). After stripping the membranes can be used for a new hybridisation to detect another factor of virulence.

Cyclic Macromolecules: Dynamics and Nonlinear Rheology, Final Report DOE Award # DE-FG02-05ER46218, Texas Tech University

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

McKenna, Gregory B.; Grubbs, Robert H.; Kornfield, Julia A.

2012-04-25

The work described in the present report had the original goal to produce large, entangled, ring polymers that were uncontaminated by linear chains and to characterize by rheological methods the dynamics of these rings. While the work fell short of this specific goal, the outcomes of the research performed under support from this grant provided novel macromolecular synthesis methods, new separation methods for ring and linear chains, and novel rheological data on bottle brush polymers, wedge polymers and dendron-based ring molecules. The grant funded a total of 8 archival manuscripts and one patent, all of which are attached to themore » present report.« less

Transport Coefficients from Large Deviation Functions

NASA Astrophysics Data System (ADS)

Gao, Chloe; Limmer, David

2017-10-01

We describe a method for computing transport coefficients from the direct evaluation of large deviation function. This method is general, relying on only equilibrium fluctuations, and is statistically efficient, employing trajectory based importance sampling. Equilibrium fluctuations of molecular currents are characterized by their large deviation functions, which is a scaled cumulant generating function analogous to the free energy. A diffusion Monte Carlo algorithm is used to evaluate the large deviation functions, from which arbitrary transport coefficients are derivable. We find significant statistical improvement over traditional Green-Kubo based calculations. The systematic and statistical errors of this method are analyzed in the context of specific transport coefficient calculations, including the shear viscosity, interfacial friction coefficient, and thermal conductivity.

Preparation of hydroxyapatite from animal bones.

PubMed

Sobczak, Agnieszka; Kowalski, Zygmunt; Wzorek, Zbigniew

2009-01-01

This paper presents the method of obtaining hydroxyapatite from animal bones. Bone sludge and calcined products were characterized by X-ray diffraction (XRD), Fourier transformed infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM). Calcium concentration was determined with titration, and phosphorus--spectrophotometrically. Making use of the AAS and ICP methods the content of microelements was determined. In all the products, hydroxyapatite was the only crystalline phase indicated. The FT-IR spectra confirmed that calcination removed the total of organic substances. Calcium and phosphorus contents were 38% and 18%, respectively, which corresponded to the Ca/P molar ratio of nonstoichiometric hydroxyapatite. The specific surfaces of products were measured by BET method. The volume of micro- and mesopores was determined.

Driver behavior profiling: An investigation with different smartphone sensors and machine learning

PubMed Central

Ferreira, Jair; Carvalho, Eduardo; Ferreira, Bruno V.; de Souza, Cleidson; Suhara, Yoshihiko; Pentland, Alex

2017-01-01

Driver behavior impacts traffic safety, fuel/energy consumption and gas emissions. Driver behavior profiling tries to understand and positively impact driver behavior. Usually driver behavior profiling tasks involve automated collection of driving data and application of computer models to generate a classification that characterizes the driver aggressiveness profile. Different sensors and classification methods have been employed in this task, however, low-cost solutions and high performance are still research targets. This paper presents an investigation with different Android smartphone sensors, and classification algorithms in order to assess which sensor/method assembly enables classification with higher performance. The results show that specific combinations of sensors and intelligent methods allow classification performance improvement. PMID:28394925

Study of trabecular bone microstructure using spatial autocorrelation analysis

NASA Astrophysics Data System (ADS)

Wald, Michael J.; Vasilic, Branimir; Saha, Punam K.; Wehrli, Felix W.

2005-04-01

The spatial autocorrelation analysis method represents a powerful, new approach to quantitative characterization of structurally quasi-periodic anisotropic materials such as trabecular bone (TB). The method is applicable to grayscale images and thus does not require any preprocessing, such as segmentation which is difficult to achieve in the limited resolution regime of in vivo imaging. The 3D autocorrelation function (ACF) can be efficiently calculated using the Fourier transform. The resulting trabecular thickness and spacing measurements are robust to the presence of noise and produce values within the expected range as determined by other methods from μCT and μMRI datasets. TB features found from the ACF are shown to correlate well with those determined by the Fuzzy Distance transform (FDT) in the transverse plane, i.e. the plane orthogonal to bone"s major axis. The method is further shown to be applicable to in-vivo μMRI data. Using the ACF, we examine data acquired in a previous study aimed at evaluating the structural implications of male hypogonadism characterized by testosterone deficiency and reduced bone mass. Specifically, we consider the hypothesis that eugonadal and hypogonadal men differ in the anisotropy of their trabecular networks. The analysis indicates a significant difference in trabecular bone thickness and longitudinal spacing between the control group and the testosterone deficient group. We conclude that spatial autocorrelation analysis is able to characterize the 3D structure and anisotropy of trabecular bone and provides new insight into the structural changes associated with osteoporotic trabecular bone loss.

The characterization and certification of a quantitative reference material for Legionella detection and quantification by qPCR.

PubMed

Baume, M; Garrelly, L; Facon, J P; Bouton, S; Fraisse, P O; Yardin, C; Reyrolle, M; Jarraud, S

2013-06-01

The characterization and certification of a Legionella DNA quantitative reference material as a primary measurement standard for Legionella qPCR. Twelve laboratories participated in a collaborative certification campaign. A candidate reference DNA material was analysed through PCR-based limiting dilution assays (LDAs). The validated data were used to statistically assign both a reference value and an associated uncertainty to the reference material. This LDA method allowed for the direct quantification of the amount of Legionella DNA per tube in genomic units (GU) and the determination of the associated uncertainties. This method could be used for the certification of all types of microbiological standards for qPCR. The use of this primary standard will improve the accuracy of Legionella qPCR measurements and the overall consistency of these measurements among different laboratories. The extensive use of this certified reference material (CRM) has been integrated in the French standard NF T90-471 (April 2010) and in the ISO Technical Specification 12 869 (Anon 2012 International Standardisation Organisation) for validating qPCR methods and ensuring the reliability of these methods. © 2013 The Society for Applied Microbiology.