Membrane filtration immobilization technique-a simple and novel method for primary isolation and enrichment of bacteriophages.

PubMed

Ghugare, G S; Nair, A; Nimkande, V; Sarode, P; Rangari, P; Khairnar, K

2017-02-01

To develop a method for the isolation and enrichment of bacteriophages selectively against specific bacteria coupled with a membrane filtration technique. Rapid isolation and concentration of host-specific bacteriophages was achieved by exposure of the sample suspected to contain bacteriophages to a specific host immobilized on a 0·45 μm membrane in a membrane filtration unit. The principle behind this method is the exploitation of host-specific interaction of bacteriophages with their host and maximizing this interaction using a classic membrane filtration method. This provides a chance for each bacteriophage in the sample to interact with the specific host on the membrane filter fitted with a vacuum pump. Specific bacteriophages of the host are retained on the membrane along with its host cells due to the effect of adsorption and these adsorbed bacteriophages (along with their hosts) on the filter disc are then amplified and enriched in regular nutritive broth tryptose soya broth by incubation. With the help of the plaque assay method, host-specific phages of various bacterial species were isolated, segregated and enriched. The phage concentration method coupled with membrane filtration immobilization of host bacteria was able to isolate and enrich the host-specific bacteriophages by several fold using a lower quantity of an environmental water sample, or other phage suspensions. Enrichment of phages from single plaques was also achieved. The isolation and detection of host-specific bacteriophages from a low density bacteriophage water sample in a single step by the use of a simple and basic microbiological technique can be achieved. Enrichment of phages from low phage titre suspensions is also achieved very effectively. © 2016 The Society for Applied Microbiology.

Detection probabilities of electrofishing, hoop nets, and benthic trawls for fishes in two western North American rivers

USGS Publications Warehouse

Smith, Christopher D.; Quist, Michael C.; Hardy, Ryan S.

2015-01-01

Research comparing different sampling techniques helps improve the efficiency and efficacy of sampling efforts. We compared the effectiveness of three sampling techniques (small-mesh hoop nets, benthic trawls, boat-mounted electrofishing) for 30 species in the Green (WY, USA) and Kootenai (ID, USA) rivers by estimating conditional detection probabilities (probability of detecting a species given its presence at a site). Electrofishing had the highest detection probabilities (generally greater than 0.60) for most species (88%), but hoop nets also had high detectability for several taxa (e.g., adult burbot Lota lota, juvenile northern pikeminnow Ptychocheilus oregonensis). Benthic trawls had low detection probabilities (<0.05) for most taxa (84%). Gear-specific effects were present for most species indicating large differences in gear effectiveness among techniques. In addition to gear effects, habitat characteristics also influenced detectability of fishes. Most species-specific habitat relationships were idiosyncratic and reflected the ecology of the species. Overall findings of our study indicate that boat-mounted electrofishing and hoop nets are the most effective techniques for sampling fish assemblages in large, coldwater rivers.

A comparison between polymerase chain reaction (PCR) and traditional techniques for the diagnosis of leptospirosis in bovines.

PubMed

Hernández-Rodríguez, Patricia; Díaz, César A; Dalmau, Ernesto A; Quintero, Gladys M

2011-01-01

Leptospirosis is caused by Leptospira, gram negative spirochaetes whose microbiologic identification is difficult due to their low rate of growth and metabolic activity. In Colombia leptospirosis diagnosis is achieved by serological techniques without unified criteria for what positive titers are. In this study we compared polymerase chain reaction (PCR) with microbiological culture and dark field microscopy for the diagnosis of leptospirosis. Microbiological and molecular techniques were performed on 83 samples of urine taken from bovines in the savannahs surrounding Bogotá in Colombia, with presumptive diagnosis of leptospirosis. 117 samples of urine taken from healthy bovines were used as negative controls. 83 samples were MAT positive with titers ≥ 1:50; 81 with titers ≥ 1:100; and 66 with titers ≥ 1:200. 36% of the total samples (73/200) were Leptospira positives by microbiological culture, 32% (63/200) by dark field microscopy and 37% (74/200) by PCR. Amplicons obtained by PCR were 482 base pair long which are Leptospira specific. An amplicon of 262 base pairs typical of pathogenic Leptospira was observed in 71 out of the 74 PCR positive samples. The remaining 3 samples showed a 240 base pair amplicon which is typical of saprophytic Leptospira. PCR as a Leptospira diagnosis technique was 100% sensitive and 99% specific in comparison to microbiological culture. Kappa value of 0.99 indicated an excellent concordance between these techniques. Sensitivity and specificity reported for MAT when compared to microbiological culture was 0.95 and 0.89 with a ≥ 1:50 cut off. PCR was a reliable method for the rapid and precise diagnosis of leptospirosis when compared to traditional techniques in our study. The research presented here will be helpful to improve diagnosis and control of leptospirosis in Colombia and other endemic countries. Copyright © 2010 Elsevier B.V. All rights reserved.

Anisotropic thermal conductivity of thin polycrystalline oxide samples

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

Tiwari, A., E-mail: abhishektiwariiitr@gmail.com; Department of Mechanical and Aerospace Engineering, Monash University, Melbourne, VIC 3800; Boussois, K.

2013-11-15

This paper reports about the development of a modified laser-flash technique and relation to measure the in-plane thermal diffusivity of thin polycrystalline oxide samples. Thermal conductivity is then calculated with the product of diffusivity, specific heat and density. Design and operating features for evaluating in-plane thermal conductivities are described. The technique is advantageous as thin samples are not glued together to measure in-plane thermal conductivities like earlier methods reported in literature. The approach was employed to study anisotropic thermal conductivity in alumina sheet, textured kaolin ceramics and montmorillonite. Since it is rare to find in-plane thermal conductivity values for suchmore » anisotropic thin samples in literature, this technique offers a useful variant to existing techniques.« less

A multiplex real-time PCR assay for identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in samples from AIDS patients with opportunistic pneumonia.

PubMed

Gago, Sara; Esteban, Cristina; Valero, Clara; Zaragoza, Oscar; Puig de la Bellacasa, Jorge; Buitrago, María José

2014-04-01

A molecular diagnostic technique based on real-time PCR was developed for the simultaneous detection of three of the most frequent causative agents of fungal opportunistic pneumonia in AIDS patients: Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii. This technique was tested in cultured strains and in clinical samples from HIV-positive patients. The methodology used involved species-specific molecular beacon probes targeted to the internal transcribed spacer regions of the rDNA. An internal control was also included in each assay. The multiplex real-time PCR assay was tested in 24 clinical strains and 43 clinical samples from AIDS patients with proven fungal infection. The technique developed showed high reproducibility (r(2) of >0.98) and specificity (100%). For H. capsulatum and Cryptococcus spp., the detection limits of the method were 20 and 2 fg of genomic DNA/20 μl reaction mixture, respectively, while for P. jirovecii the detection limit was 2.92 log10 copies/20 μl reaction mixture. The sensitivity in vitro was 100% for clinical strains and 90.7% for clinical samples. The assay was positive for 92.5% of the patients. For one of the patients with proven histoplasmosis, P. jirovecii was also detected in a bronchoalveolar lavage sample. No PCR inhibition was detected. This multiplex real-time PCR technique is fast, sensitive, and specific and may have clinical applications.

Principles, Techniques, and Applications of Tissue Microfluidics

NASA Technical Reports Server (NTRS)

Wade, Lawrence A.; Kartalov, Emil P.; Shibata, Darryl; Taylor, Clive

2011-01-01

The principle of tissue microfluidics and its resultant techniques has been applied to cell analysis. Building microfluidics to suit a particular tissue sample would allow the rapid, reliable, inexpensive, highly parallelized, selective extraction of chosen regions of tissue for purposes of further biochemical analysis. Furthermore, the applicability of the techniques ranges beyond the described pathology application. For example, they would also allow the posing and successful answering of new sets of questions in many areas of fundamental research. The proposed integration of microfluidic techniques and tissue slice samples is called "tissue microfluidics" because it molds the microfluidic architectures in accordance with each particular structure of each specific tissue sample. Thus, microfluidics can be built around the tissues, following the tissue structure, or alternatively, the microfluidics can be adapted to the specific geometry of particular tissues. By contrast, the traditional approach is that microfluidic devices are structured in accordance with engineering considerations, while the biological components in applied devices are forced to comply with these engineering presets.

Sampling protocol for monitoring abiotic and biotic characteristics of mountain ponds and lakes

USGS Publications Warehouse

Hoffman, Robert L.; Tyler, Torrey J.; Larson, Gary L.; Adams, Michael J.; Wente, Wendy; Galvan, Stephanie

2005-01-01

This document describes field techniques and procedures used for sampling mountain ponds and lakes. These techniques and procedures will be used primarily to monitor, as part of long-term programs in National Parks and other protected areas, the abiotic and biotic characteristics of naturally occurring permanent montane lentic systems up to 75 ha in surface area. However, the techniques and procedures described herein also can be used to sample temporary or ephemeral montane lentic sites. Each Standard Operating Procedure (SOP) section addresses a specific component of the limnological investigation, and describes in detail field sampling methods pertaining to parameters to be measured for each component.

COMPARISON OF A GENUS-SPECIFIC CONVENTIONAL PCR AND A SPECIES-SPECIFIC NESTED-PCR FOR MALARIA DIAGNOSIS USING FTA COLLECTED SAMPLES FROM KINGDOM OF SAUDI ARABIA.

PubMed

Al-Harthi, Saeed A

2015-12-01

Molecular tools are increasingly accepted as the most sensitive and reliable techniques for malaria diagnosis and epidemiological surveys. Also, collection of finger prick blood spots onto filter papers is the most simple and affordable method for samples preservation and posterior molecular analysis, especially in rural endemic regions where malaria remains a major health problem. Two malaria molecular diagnostic tests, a Plasmodium genus-specific conventional PCR and a Plasmodium species-specific Nested PCR, were evaluated using DNA templates prepared from Whatman-FTA cards' dry blood spots using both, Methanol-fixation/Heat-extraction and FTA commercial purification kit. A total of 121 blood samples were collected from six Saudi south-western endemic districts both, as thick and thin films for routine microscopic screening and onto FTA cards for molecular studies. Out of the 121 samples, 75 were P. falciparum positive by at least one technique. No other species of Plasmodium were detected. P. falciparum parasites were identified in 69/75 (92%) samples by microscopic screening in health care centers. P. genus-specific PCR was able to amplify P. falciparum DNA in 41/75 (55%) and 59/75 (79%) samples using Methanol-fixation/Heat-extraction and FTA purification kit, respectively. P. species-specific Nested PCR revealed 68/75 (91%) and 75/75 (100%) positive samples using DNA templates were isolated by Methanol-fixation/Heat- extraction and FTA purification methods, respectively. The species-specific Nested PCR applied to Whatman-FTA preserved and processed blood samples represents the best alternative to classical microscopy for malaria diagnosis, particularly in epidemiological screening.

Accuracy of a new clean-catch technique for diagnosis of urinary tract infection in infants younger than 90 days of age

PubMed Central

Herreros, María Luisa; Tagarro, Alfredo; García-Pose, Araceli; Sánchez, Aida; Cañete, Alfonso; Gili, Pablo

2015-01-01

OBJECTIVE: To evaluate the accuracy of diagnosing urinary tract infections using a new, recently described, standardized clean-catch collection technique. METHODS: Cross-sectional study of infants <90 days old admitted due to fever without a source, with two matched samples of urine obtained using two different methods: clean-catch standardized stimulation technique and bladder catheterization. RESULTS: Sixty paired urine cultures were obtained. The median age was 44-days-old. Seventeen percent were male infants. Clean-catch technique sensitivity was 97% (95% CI 82% to 100%) and specificity was 89% (95% CI 65% to 98%). The contamination rate of clean-catch samples was lower (5%) than the contamination rate of catheter specimens (8%). CONCLUSIONS: The sensitivity and specificity of urine cultures obtained using the clean-catch method through the new technique were accurate and the contamination rate was low. These results suggest that this technique is a valuable, alternative method for urinary tract infection diagnosis. PMID:26435675

Identification of Explosives from Porous Materials: Applications Using Reverse Phase High Performance Liquid Chromatography and Gas Chromatography

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

C.J. Miller; G. Elias; N.C. Schmitt

2010-06-01

High performance liquid chromatography and gas chromatography techniques are well documented and widely used for the detection of trace explosives from organic solvents. These techniques were modified to specifically identify and quantify explosives extracted from various materials taken from people who had recently handled explosives. Documented techniques were modified to specifically detect and quantify RDX, TNT, and PETN from denim, colored flannel, vinyl, and canvas extracted in methanol using no sample cleanup prior to analysis. The methanol extracts were injected directly into several different column types and analyzed by HPLC-UV and/or GC-ECD. This paper describes general screening methods that weremore » used to determine the presence of explosives in unknown samples and techniques that have been optimized for quantification of each explosive from the substrate extracts.« less

Spectroscopic diagnostics for bacteria in biologic sample

DOEpatents

El-Sayed, Mostafa A.; El-Sayed, Ivan H.

2002-01-01

A method to analyze and diagnose specific bacteria in a biologic sample using spectroscopy is disclosed. The method includes obtaining the spectra of a biologic sample of a non-infected patient for use as a reference, subtracting the reference from the spectra of an infected sample, and comparing the fingerprint regions of the resulting differential spectrum with reference spectra of bacteria in saline. Using this diagnostic technique, specific bacteria can be identified sooner and without culturing, bacteria-specific antibiotics can be prescribed sooner, resulting in decreased likelihood of antibiotic resistance and an overall reduction of medical costs.

Quantification of ferritin bound iron in human serum using species-specific isotope dilution mass spectrometry.

PubMed

Ren, Yao; Walczyk, Thomas

2014-09-01

Ferritin is a hollow sphere protein composed of 24 subunits that can store up to 4500 iron atoms in its inner cavity. It is mainly found in the liver and spleen but also in serum at trace levels. Serum ferritin is considered as the best single indicator in assessing body iron stores except liver or bone marrow biopsy. However, it is confounded by other disease conditions. Ferritin bound iron (FBI) and ferritin saturation have been suggested as more robust biomarkers. The current techniques for FBI determination are limited by low antibody specificity, low instrument sensitivity and possible analyte losses during sample preparation. The need for a highly sensitive and reliable method is widely recognized. Here we describe a novel technique to detect serum FBI using species-specific isotope dilution mass spectrometry (SS-IDMS). [(57)Fe]-ferritin was produced by biosynthesis and in vitro labeling with the (57)Fe spike in the form of [(57)Fe]-citrate after cell lysis and heat treatment. [(57)Fe]-ferritin for sample spiking was further purified by fast liquid protein chromatography. Serum ferritin and added [(57)Fe]-ferritin were separated from other iron species by ultrafiltration followed by isotopic analysis of FBI using negative thermal ionization mass spectrometry. Repeatability of our assay is 8% with an absolute detection limit of 18 ng FBI in the sample. As compared to other speciation techniques, SS-IDMS offers maximum control over sample losses and species conversion during analysis. The described technique may therefore serve as a reference technique for clinical applications of FBI as a new biomarker for assessing body iron status.

Estimating the sensitivity and specificity of Kato-Katz stool examination technique for detection of hookworms, Ascaris lumbricoides and Trichuris trichiura infections in humans in the absence of a 'gold standard'.

PubMed

Tarafder, M R; Carabin, H; Joseph, L; Balolong, E; Olveda, R; McGarvey, S T

2010-03-15

The accuracy of the Kato-Katz technique in identifying individuals with soil-transmitted helminth (STH) infections is limited by day-to-day variation in helminth egg excretion, confusion with other parasites and the laboratory technicians' experience. We aimed to estimate the sensitivity and specificity of the Kato-Katz technique to detect infection with Ascaris lumbricoides, hookworm and Trichuris trichiura using a Bayesian approach in the absence of a 'gold standard'. Data were obtained from a longitudinal study conducted between January 2004 and December 2005 in Samar Province, the Philippines. Each participant provided between one and three stool samples over consecutive days. Stool samples were examined using the Kato-Katz technique and reported as positive or negative for STHs. In the presence of measurement error, the true status of each individual is considered as latent data. Using a Bayesian method, we calculated marginal posterior densities of sensitivity and specificity parameters from the product of the likelihood function of observed and latent data. A uniform prior distribution was used (beta distribution: alpha=1, beta=1). A total of 5624 individuals provided at least one stool sample. One, two and three stool samples were provided by 1582, 1893 and 2149 individuals, respectively. All STHs showed variation in test results from day to day. Sensitivity estimates of the Kato-Katz technique for one stool sample were 96.9% (95% Bayesian Credible Interval [BCI]: 96.1%, 97.6%), 65.2% (60.0%, 69.8%) and 91.4% (90.5%, 92.3%), for A. lumbricoides, hookworm and T. trichiura, respectively. Specificity estimates for one stool sample were 96.1% (95.5%, 96.7%), 93.8% (92.4%, 95.4%) and 94.4% (93.2%, 95.5%), for A. lumbricoides, hookworm and T. trichiura, respectively. Our results show that the Kato-Katz technique can perform with reasonable accuracy with one day's stool collection for A. lumbricoides and T. trichiura. Low sensitivity of the Kato-Katz for detection of hookworm infection may be related to rapid degeneration of delicate hookworm eggs with time. (c) 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Methodological integrative review of the work sampling technique used in nursing workload research.

PubMed

Blay, Nicole; Duffield, Christine M; Gallagher, Robyn; Roche, Michael

2014-11-01

To critically review the work sampling technique used in nursing workload research. Work sampling is a technique frequently used by researchers and managers to explore and measure nursing activities. However, work sampling methods used are diverse making comparisons of results between studies difficult. Methodological integrative review. Four electronic databases were systematically searched for peer-reviewed articles published between 2002-2012. Manual scanning of reference lists and Rich Site Summary feeds from contemporary nursing journals were other sources of data. Articles published in the English language between 2002-2012 reporting on research which used work sampling to examine nursing workload. Eighteen articles were reviewed. The review identified that the work sampling technique lacks a standardized approach, which may have an impact on the sharing or comparison of results. Specific areas needing a shared understanding included the training of observers and subjects who self-report, standardization of the techniques used to assess observer inter-rater reliability, sampling methods and reporting of outcomes. Work sampling is a technique that can be used to explore the many facets of nursing work. Standardized reporting measures would enable greater comparison between studies and contribute to knowledge more effectively. Author suggestions for the reporting of results may act as guidelines for researchers considering work sampling as a research method. © 2014 John Wiley & Sons Ltd.

A preclustering-based ensemble learning technique for acute appendicitis diagnoses.

PubMed

Lee, Yen-Hsien; Hu, Paul Jen-Hwa; Cheng, Tsang-Hsiang; Huang, Te-Chia; Chuang, Wei-Yao

2013-06-01

Acute appendicitis is a common medical condition, whose effective, timely diagnosis can be difficult. A missed diagnosis not only puts the patient in danger but also requires additional resources for corrective treatments. An acute appendicitis diagnosis constitutes a classification problem, for which a further fundamental challenge pertains to the skewed outcome class distribution of instances in the training sample. A preclustering-based ensemble learning (PEL) technique aims to address the associated imbalanced sample learning problems and thereby support the timely, accurate diagnosis of acute appendicitis. The proposed PEL technique employs undersampling to reduce the number of majority-class instances in a training sample, uses preclustering to group similar majority-class instances into multiple groups, and selects from each group representative instances to create more balanced samples. The PEL technique thereby reduces potential information loss from random undersampling. It also takes advantage of ensemble learning to improve performance. We empirically evaluate this proposed technique with 574 clinical cases obtained from a comprehensive tertiary hospital in southern Taiwan, using several prevalent techniques and a salient scoring system as benchmarks. The comparative results show that PEL is more effective and less biased than any benchmarks. The proposed PEL technique seems more sensitive to identifying positive acute appendicitis than the commonly used Alvarado scoring system and exhibits higher specificity in identifying negative acute appendicitis. In addition, the sensitivity and specificity values of PEL appear higher than those of the investigated benchmarks that follow the resampling approach. Our analysis suggests PEL benefits from the more representative majority-class instances in the training sample. According to our overall evaluation results, PEL records the best overall performance, and its area under the curve measure reaches 0.619. The PEL technique is capable of addressing imbalanced sample learning associated with acute appendicitis diagnosis. Our evaluation results suggest PEL is less biased toward a positive or negative class than the investigated benchmark techniques. In addition, our results indicate the overall effectiveness of the proposed technique, compared with prevalent scoring systems or salient classification techniques that follow the resampling approach. Copyright © 2013 Elsevier B.V. All rights reserved.

Comparison of Diagnostic Techniques for Helicobacter cetorum Infection in Wild Atlantic Bottlenose Dolphins (Tursiops truncatus)

PubMed Central

Harper, Claudia G.; Whary, Mark T.; Feng, Yan; Rhinehart, Howard L.; Wells, Randall S.; Xu, Shilu; Taylor, Nancy S.; Fox, James G.

2003-01-01

Helicobacter cetorum sp. nov. has been cultured from the stomach of Atlantic white-sided dolphins (Lagenorhynchus acutus) and the feces of Pacific white-sided (L. obliquidens) and Atlantic bottlenose (Tursiops truncatus) dolphins and a beluga whale (Delphinapterus leucas). H. cetorum has high homology to Helicobacter pylori as shown by 16S rRNA sequencing, and H. cetorum infection has been associated with gastritis and clinical signs in cetaceans. Because the prevalence of H. cetorum in wild populations is unknown, minimally invasive techniques for detecting H. cetorum were compared for 20 wild bottlenose dolphins sampled as part of a long-term health study. Fecal samples were tested for helicobacter by culture, Southern blotting, and PCR using genus-specific and H. cetorum-specific primers. An enzyme-linked immunosorbent assay (ELISA) was developed to measure H. cetorum immunoglobulin G (IgG). H. cetorum was cultured from 4 of 20 fecal samples, 7 samples were positive using Helicobacter sp. PCR, and 8 samples were positive for H. cetorum using species-specific primers. Two additional fecal samples were positive by Helicobacter sp. Southern blotting, suggesting infection with another helicobacter. All 20 sera contained high levels of IgG antibodies to H. cetorum that were significantly lowered by preabsorption of the sera with whole-cell suspensions of H. cetorum (P < 0.02). Until the specificity of the serum ELISA can be determined by testing sera from dolphins confirmed to be uninfected, PCR and Southern blot screenings of feces are the most sensitive techniques for detection of H. cetorum, and results indicate there is at least a 50% prevalence of H. cetorum infection in these dolphins. PMID:12843010

Principles, Techniques, and Applications of Tissue Microfluidics

NASA Technical Reports Server (NTRS)

Wade, Lawrence A.; Kartalov, Emil P.; Shibata, Darryl; Taylor, Clive

2011-01-01

The principle of tissue microfluidics and its resultant techniques has been applied to cell analysis. Building microfluidics to suit a particular tissue sample would allow the rapid, reliable, inexpensive, highly parallelized, selective extraction of chosen regions of tissue for purposes of further biochemical analysis. Furthermore, the applicability of the techniques ranges beyond the described pathology application. For example, they would also allow the posing and successful answering of new sets of questions in many areas of fundamental research. The proposed integration of microfluidic techniques and tissue slice samples is called tissue microfluidics because it molds the microfluidic architectures in accordance with each particular structure of each specific tissue sample. Thus, microfluidics can be built around the tissues, following the tissue structure, or alternatively, the microfluidics can be adapted to the specific geometry of particular tissues. By contrast, the traditional approach is that microfluidic devices are structured in accordance with engineering considerations, while the biological components in applied devices are forced to comply with these engineering presets. The proposed principles represent a paradigm shift in microfluidic technology in three important ways: Microfluidic devices are to be directly integrated with, onto, or around tissue samples, in contrast to the conventional method of off-chip sample extraction followed by sample insertion in microfluidic devices. Architectural and operational principles of microfluidic devices are to be subordinated to suit specific tissue structure and needs, in contrast to the conventional method of building devices according to fluidic function alone and without regard to tissue structure. Sample acquisition from tissue is to be performed on-chip and is to be integrated with the diagnostic measurement within the same device, in contrast to the conventional method of off-chip sample prep and subsequent insertion into a diagnostic device. A more advanced form of tissue integration with microfluidics is tissue encapsulation, wherein the sample is completely encapsulated within a microfluidic device, to allow for full surface access. The immediate applications of these approaches lie with diagnostics of tissue slices and biopsy samples e.g. for cancer but the approaches would also be very useful in comparative genomics and other areas of fundamental research involving heterogeneous tissue samples.

MRI and unilateral NMR study of reindeer skin tanning processes.

PubMed

Zhu, Lizheng; Del Federico, Eleonora; Ilott, Andrew J; Klokkernes, Torunn; Kehlet, Cindie; Jerschow, Alexej

2015-04-07

The study of arctic or subarctic indigenous skin clothing material, known for its design and ability to keep the body warm, provides information about the tanning materials and techniques. The study also provides clues about the culture that created it, since tanning processes are often specific to certain indigenous groups. Untreated skin samples and samples treated with willow (Salix sp) bark extract and cod liver oil are compared in this study using both MRI and unilateral NMR techniques. The two types of samples show different proton spatial distributions and different relaxation times, which may also provide information about the tanning technique and aging behavior.

Uranium determination in natural water by the fissiontrack technique

USGS Publications Warehouse

Reimer, G.M.

1975-01-01

The fission track technique, utilizing the neutron-induced fission of uranium-235, provides a versatile analytical method for the routine analysis of uranium in liquid samples of natural water. A detector is immersed in the sample and both are irradiated. The fission track density observed in the detector is directly proportional to the uranium concentration. The specific advantages of this technique are: (1) only a small quantity of sample, typically 0.1-1 ml, is needed; (2) no sample concentration is necessary; (3) it is capable of providing analyses with a lower reporting limit of 1 ??g per liter; and (4) the actual time spent on an analysis can be only a few minutes. This paper discusses and describes the method. ?? 1975.

Psychometric Evaluation of Lexical Diversity Indices: Assessing Length Effects

ERIC Educational Resources Information Center

Fergadiotis, Gerasimos; Wright, Heather Harris; Green, Samuel B.

2015-01-01

Purpose: Several novel techniques have been developed recently to assess the breadth of a speaker's vocabulary exhibited in a language sample. The specific aim of this study was to increase our understanding of the validity of the scores generated by different lexical diversity (LD) estimation techniques. Four techniques were explored: D, Maas,…

Reproducibility of combinatorial peptide ligand libraries for proteome capture evaluated by selected reaction monitoring.

PubMed

Di Girolamo, Francesco; Righetti, Pier Giorgio; Soste, Martin; Feng, Yuehan; Picotti, Paola

2013-08-26

Systems biology studies require the capability to quantify with high precision proteins spanning a broad range of abundances across multiple samples. However, the broad range of protein expression in cells often precludes the detection of low-abundance proteins. Different sample processing techniques can be applied to increase proteome coverage. Among these, combinatorial (hexa)peptide ligand libraries (CPLLs) bound to solid matrices have been used to specifically capture and detect low-abundance proteins in complex samples. To assess whether CPLL capture can be applied in systems biology studies involving the precise quantitation of proteins across a multitude of samples, we evaluated its performance across the whole range of protein abundances in Saccharomyces cerevisiae. We used selected reaction monitoring assays for a set of target proteins covering a broad abundance range to quantitatively evaluate the precision of the approach and its capability to detect low-abundance proteins. Replicated CPLL-isolates showed an average variability of ~10% in the amount of the isolated proteins. The high reproducibility of the technique was not dependent on the abundance of the protein or the amount of beads used for the capture. However, the protein-to-bead ratio affected the enrichment of specific proteins. We did not observe a normalization effect of CPLL beads on protein abundances. However, CPLLs enriched for and depleted specific sets of proteins and thus changed the abundances of proteins from a whole proteome extract. This allowed the identification of ~400 proteins otherwise undetected in an untreated sample, under the experimental conditions used. CPLL capture is thus a useful tool to increase protein identifications in proteomic experiments, but it should be coupled to the analysis of untreated samples, to maximize proteome coverage. Our data also confirms that CPLL capture is reproducible and can be confidently used in quantitative proteomic experiments. Combinatorial hexapeptide ligand libraries (CPLLs) bound to solid matrices have been proposed to specifically capture and detect low-abundance proteins in complex samples. To assess whether the CPLL capture can be confidently applied in systems biology studies involving the precise quantitation of proteins across a broad range of abundances and a multitude of samples, we evaluated its reproducibility and performance features. Using selected reaction monitoring assays for proteins covering the whole range of abundances we show that the technique is reproducible and compatible with quantitative proteomic studies. However, the protein-to-bead ratio affects the enrichment of specific proteins and CPLLs depleted specific sets of proteins from a whole proteome extract. Our results suggest that CPLL-based analyses should be coupled to the analysis of untreated samples, to maximize proteome coverage. Overall, our data confirms the applicability of CPLLs in systems biology research and guides the correct use of this technique. Copyright © 2013 Elsevier B.V. All rights reserved.

3D imaging of cells and tissues by focused ion beam/scanning electron microscopy (FIB/SEM).

PubMed

Drobne, Damjana

2013-01-01

Integration of a scanning electron microscope (SEM) and focused ion beam (FIB) technology into a single FIB/SEM system permits use of the FIB as a nano-scalpel to reveal site-specific subsurface microstructures which can be examined in great detail by SEM. The FIB/SEM technology is widely used in the semiconductor industry and material sciences, and recently its use in the life sciences has been initiated. Samples for FIB/SEM investigation can be either embedded in a plastic matrix, the traditional means of preparation of transmission electron microscopy (TEM) specimens, or simply dried as in samples prepared for SEM imaging. Currently, FIB/SEM is used in the life sciences for (a) preparation by the lift-out technique of lamella for TEM analysis, (b) tomography of samples embedded in a matrix, and (c) in situ site-specific FIB milling and SEM imaging using a wide range of magnifications. Site-specific milling and imaging has attracted wide interest as a technique in structural research of single eukaryotic and prokaryotic cells, small animals, and different animal tissue, but it still remains to be explored more thoroughly. In the past, preparation of samples for site-specific milling and imaging by FIB/SEM has typically adopted the embedding techniques used for TEM samples, and which have been very well described in the literature. Sample preparation protocols for the use of dried samples in FIB/SEM have been less well investigated. The aim of this chapter is to encourage application of FIB/SEM on dried biological samples. A detailed description of conventional dried sample preparation and FIB/SEM investigation of dried biological samples is presented. The important steps are described and illustrated, and direct comparison between embedded and dried samples of same tissues is provided. The ability to discover links between gross morphology of the tissue or organ, surface characteristics of any selected region, and intracellular structural details on the nanometer scale is an appealing application of electron microscopy in the life sciences and merits further exploration.

Carbon-13 and proton nuclear magnetic resonance analysis of shale-derived refinery products and jet fuels and of experimental referee broadened-specification jet fuels

NASA Technical Reports Server (NTRS)

Dalling, D. K.; Bailey, B. K.; Pugmire, R. J.

1984-01-01

A proton and carbon-13 nuclear magnetic resonance (NMR) study was conducted of Ashland shale oil refinery products, experimental referee broadened-specification jet fuels, and of related isoprenoid model compounds. Supercritical fluid chromatography techniques using carbon dioxide were developed on a preparative scale, so that samples could be quantitatively separated into saturates and aromatic fractions for study by NMR. An optimized average parameter treatment was developed, and the NMR results were analyzed in terms of the resulting average parameters; formulation of model mixtures was demonstrated. Application of novel spectroscopic techniques to fuel samples was investigated.

Analysis of the site-specific carbon isotope composition of propane by gas source isotope ratio mass spectrometer

NASA Astrophysics Data System (ADS)

Piasecki, Alison; Sessions, Alex; Lawson, Michael; Ferreira, A. A.; Neto, E. V. Santos; Eiler, John M.

2016-09-01

Site-specific isotope ratio measurements potentially provide valuable information about the formation and degradation of complex molecules-information that is lost in conventional bulk isotopic measurements. Here we discuss the background and possible applications of such measurements, and present a technique for studying the site-specific carbon isotope composition of propane at natural abundance based on mass spectrometric analysis of the intact propane molecule and its fragment ions. We demonstrate the feasibility of this approach through measurements of mixtures of natural propane and propane synthesized with site-specific 13C enrichment, and we document the limits of precision of our technique. We show that mass balance calculations of the bulk δ13C of propane based on our site-specific measurements is generally consistent with independent constraints on bulk δ13C. We further demonstrate the accuracy of the technique, and illustrate one of its simpler applications by documenting the site-specific carbon isotope signature associated with gas phase diffusion of propane, confirming that our measurements conform to the predictions of the kinetic theory of gases. This method can be applied to propane samples of moderate size (tens of micromoles) isolated from natural gases. Thus, it provides a means of studying the site-specific stable isotope systematics of propane at natural isotope abundances on sample sizes that are readily recovered from many natural environments. This method may also serve as a model for future techniques that apply high-resolution mass spectrometry to study the site-specific isotopic distributions of larger organic molecules, with potential applications to biosynthesis, forensics and other geochemical subjects.

A fast method for detecting Cryptosporidium parvum oocysts in real world samples

NASA Astrophysics Data System (ADS)

Stewart, Shona; McClelland, Lindy; Maier, John

2005-04-01

Contamination of drinking water with pathogenic microorganisms such as Cryptosporidium has become an increasing concern in recent years. Cryptosporidium oocysts are particularly problematic, as infections caused by this organism can be life threatening in immunocompromised patients. Current methods for monitoring and analyzing water are often laborious and require experts to conduct. In addition, many of the techniques require very specific reagents to be employed. These factors add considerable cost and time to the analytical process. Raman spectroscopy provides specific molecular information on samples, and offers advantages of speed, sensitivity and low cost over current methods of water monitoring. Raman spectroscopy is an optical method that has demonstrated the capability to identify and differentiate microorganisms at the species and strain levels. In addition, this technique has exhibited sensitivities down to the single organism detection limit. We have employed Raman spectroscopy and Raman Chemical Imaging, in conjunction with chemometric techniques, to detect small numbers of oocysts in the presence of interferents derived from real-world water samples. Our investigations have also indicated that Raman Chemical Imaging may provide chemical and physiological information about an oocyst sample which complements information provided by the traditional methods. This work provides evidence that Raman imaging is a useful technique for consideration in the water quality industry.

Veterinary extension on sampling techniques related to heartwater research.

PubMed

Steyn, H C; McCrindle, C M E; Du Toit, D

2010-09-01

Heartwater, a tick-borne disease caused by Ehrlichia ruminantium, is considered to be a significant cause of mortality amongst domestic and wild ruminants in South Africa. The main vector is Amblyomma hebraeum and although previous epidemiological studies have outlined endemic areas based on mortalities, these have been limited by diagnostic methods which relied mainly on positive brain smears. The indirect fluorescent antibody test (IFA) has a low specificity for heartwater organisms as it cross-reacts with some other species. Since the advent of biotechnology and genomics, molecular epidemiology has evolved using the methodology of traditional epidemiology coupled with the new molecular techniques. A new quantitative real-time polymerase chain reaction (qPCR) test has been developed for rapid and accurate diagnosis of heartwater in the live animal. This method can also be used to survey populations of A. hebraeum ticks for heartwater. Sampling whole blood and ticks for this qPCR differs from routine serum sampling, which is used for many serological tests. Veterinary field staff, particularly animal health technicians, are involved in surveillance and monitoring of controlled and other diseases of animals in South Africa. However, it was found that the sampling of whole blood was not done correctly, probably because it is a new sampling technique specific for new technology, where the heartwater organism is much more labile than the serum antibodies required for other tests. This qPCR technique is highly sensitive and can diagnose heartwater in the living animal within 2 hours, in time to treat it. Poor sampling techniques that decrease the sensitivity of the test will, however, result in a false negative diagnosis. This paper describes the development of a skills training programme for para-veterinary field staff, to facilitate research into the molecular epidemiology of heartwater in ruminants and eliminate any sampling bias due to collection errors. Humane handling techniques were also included in the training, in line with the current focus on improved livestock welfare.

Psychometric Evaluation of Lexical Diversity Indices: Assessing Length Effects.

PubMed

Fergadiotis, Gerasimos; Wright, Heather Harris; Green, Samuel B

2015-06-01

Several novel techniques have been developed recently to assess the breadth of a speaker's vocabulary exhibited in a language sample. The specific aim of this study was to increase our understanding of the validity of the scores generated by different lexical diversity (LD) estimation techniques. Four techniques were explored: D, Maas, measure of textual lexical diversity, and moving-average type-token ratio. Four LD indices were estimated for language samples on 4 discourse tasks (procedures, eventcasts, story retell, and recounts) from 442 adults who are neurologically intact. The resulting data were analyzed using structural equation modeling. The scores for measure of textual lexical diversity and moving-average type-token ratio were stronger indicators of the LD of the language samples. The results for the other 2 techniques were consistent with the presence of method factors representing construct-irrelevant sources. These findings offer a deeper understanding of the relative validity of the 4 estimation techniques and should assist clinicians and researchers in the selection of LD measures of language samples that minimize construct-irrelevant sources.

Decomposition techniques

USGS Publications Warehouse

Chao, T.T.; Sanzolone, R.F.

1992-01-01

Sample decomposition is a fundamental and integral step in the procedure of geochemical analysis. It is often the limiting factor to sample throughput, especially with the recent application of the fast and modern multi-element measurement instrumentation. The complexity of geological materials makes it necessary to choose the sample decomposition technique that is compatible with the specific objective of the analysis. When selecting a decomposition technique, consideration should be given to the chemical and mineralogical characteristics of the sample, elements to be determined, precision and accuracy requirements, sample throughput, technical capability of personnel, and time constraints. This paper addresses these concerns and discusses the attributes and limitations of many techniques of sample decomposition along with examples of their application to geochemical analysis. The chemical properties of reagents as to their function as decomposition agents are also reviewed. The section on acid dissolution techniques addresses the various inorganic acids that are used individually or in combination in both open and closed systems. Fluxes used in sample fusion are discussed. The promising microwave-oven technology and the emerging field of automation are also examined. A section on applications highlights the use of decomposition techniques for the determination of Au, platinum group elements (PGEs), Hg, U, hydride-forming elements, rare earth elements (REEs), and multi-elements in geological materials. Partial dissolution techniques used for geochemical exploration which have been treated in detail elsewhere are not discussed here; nor are fire-assaying for noble metals and decomposition techniques for X-ray fluorescence or nuclear methods be discussed. ?? 1992.

Data in support of the detection of genetically modified organisms (GMOs) in food and feed samples.

PubMed

Alasaad, Noor; Alzubi, Hussein; Kader, Ahmad Abdul

2016-06-01

Food and feed samples were randomly collected from different sources, including local and imported materials from the Syrian local market. These included maize, barley, soybean, fresh food samples and raw material. GMO detection was conducted by PCR and nested PCR-based techniques using specific primers for the most used foreign DNA commonly used in genetic transformation procedures, i.e., 35S promoter, T-nos, epsps, cryIA(b) gene and nptII gene. The results revealed for the first time in Syria the presence of GM foods and feeds with glyphosate-resistant trait of P35S promoter and NOS terminator in the imported soybean samples with high frequency (5 out of the 6 imported soybean samples). While, tests showed negative results for the local samples. Also, tests revealed existence of GMOs in two imported maize samples detecting the presence of 35S promoter and nos terminator. Nested PCR results using two sets of primers confirmed our data. The methods applied in the brief data are based on DNA analysis by Polymerase Chain Reaction (PCR). This technique is specific, practical, reproducible and sensitive enough to detect up to 0.1% GMO in food and/or feedstuffs. Furthermore, all of the techniques mentioned are economic and can be applied in Syria and other developing countries. For all these reasons, the DNA-based analysis methods were chosen and preferred over protein-based analysis.

Development of combination tapered fiber-optic biosensor dip probe for quantitative estimation of interleukin-6 in serum samples

NASA Astrophysics Data System (ADS)

Wang, Chun Wei; Manne, Upender; Reddy, Vishnu B.; Oelschlager, Denise K.; Katkoori, Venkat R.; Grizzle, William E.; Kapoor, Rakesh

2010-11-01

A combination tapered fiber-optic biosensor (CTFOB) dip probe for rapid and cost-effective quantification of proteins in serum samples has been developed. This device relies on diode laser excitation and a charged-coupled device spectrometer and functions on a technique of sandwich immunoassay. As a proof of principle, this technique was applied in a quantitative estimation of interleukin IL-6. The probes detected IL-6 at picomolar levels in serum samples obtained from a patient with lupus, an autoimmune disease, and a patient with lymphoma. The estimated concentration of IL-6 in the lupus sample was 5.9 +/- 0.6 pM, and in the lymphoma sample, it was below the detection limit. These concentrations were verified by a procedure involving bead-based xMAP technology. A similar trend in the concentrations was observed. The specificity of the CTFOB dip probes was assessed by analysis with receiver operating characteristics. This analysis suggests that the dip probes can detect 5-pM or higher concentration of IL-6 in these samples with specificities of 100%. The results provide information for guiding further studies in the utilization of these probes to quantify other analytes in body fluids with high specificity and sensitivity.

Analytical techniques for identification and study of organic matter in returned lunar samples

NASA Technical Reports Server (NTRS)

Burlingame, A. L.

1974-01-01

The results of geochemical research are reviewed. Emphasis is placed on the contribution of mass spectrometric data to the solution of specific structural problems. Information on the mass spectrometric behavior of compounds of geochemical interest is reviewed and currently available techniques of particular importance to geochemistry, such as gas chromatograph-mass spectrometer coupling, modern sample introduction methods, and computer application in high resolution mass spectrometry, receive particular attention.

Evaluation of a new serological technique for detecting rabies virus antibodies following vaccination.

PubMed

Ma, Xiaoyue; Niezgoda, Michael; Blanton, Jesse D; Recuenco, Sergio; Rupprecht, Charles E

2012-08-03

Two major techniques are currently used to estimate rabies virus antibody values: neutralization assays, such as the rapid fluorescent focus inhibition test (RFFIT), and enzyme-linked immunosorbent assays (ELISAs). The RFFIT is considered the gold standard assay and has been used to assess the titer of rabies virus neutralizing antibodies for more than three decades. In the late 1970s, ELISA began to be used to estimate the level of rabies virus antibody and has recently been used by some laboratories as an alternate screening test for animal sera. Although the ELISA appears simpler, safer and more efficient, the assay is less sensitive in detecting low values of rabies virus neutralizing antibodies than neutralization tests. This study was designed to evaluate a new ELISA-based method for detecting rabies virus binding antibody. This new technique uses electro-chemi-luminescence labels and carbon electrode plates to detect binding events. In this comparative study, the RFFIT and the new ELISA-based technique were used to evaluate the level of rabies virus antibodies in human and animal serum samples. By using a conservative approximation of 0.15 IU/ml as a cutoff point, the new ELISA-based technique demonstrated a sensitivity of 100% and a specificity of 95% for human samples and for experimental animal samples. The sensitivity and specificity for field animal samples was 96% and 95%, respectively. The preliminary results from this study appear promising and demonstrate a higher sensitivity than traditional ELISA methods. Published by Elsevier Ltd.

Specificity of coliphages in evaluating marker efficacy: a new insight for water quality indicators.

PubMed

Mookerjee, Subham; Batabyal, Prasenjit; Halder, Madhumanti; Palit, Anup

2014-11-01

Conventional procedures for qualitative assessment of coliphage are time consuming multiple step approach for achieving results. A modified and rapid technique has been introduced for determination of coliphage contamination among potable water sources during water borne outbreaks. During December 2013, 40 water samples from different potable water sources, were received for water quality analyses, from a jaundice affected Municipality of West Bengal, India. Altogether, 30% water samples were contaminated with coliform (1-20 cfu/ml) and 5% with E. coli (2-5 cfu/ml). Among post-outbreak samples, preponderance of coliform has decreased (1-4 cfu/ml) with total absence of E. coli. While standard technique has detected 55% outbreak samples with coliphage contamination, modified technique revealed that 80%, double than that of bacteriological identification rate, were contaminated with coliphages (4-20 pfu/10 ml). However, post-outbreak samples were detected with 1-5 pfu/10 ml coliphages among 20% samples. Coliphage detection rate through modified technique was nearly double (50%) than that of standard technique (27.5%). In few samples (with coliform load of 10-100 cfu/ml), while modified technique could detect coliphages among six samples (10-20 pfu/10 ml), standard protocol failed to detect coliphage in any of them. An easy, rapid and accurate modified technique has thereby been implemented for coliphage assessment from water samples. Coliform free water does not always signify pathogen free potable water and it is demonstrated that coliphage is a more reliable 'biomarker' to ascertain contamination level in potable water. Copyright © 2014 Elsevier B.V. All rights reserved.

K-Fold Crossvalidation in Canonical Analysis.

ERIC Educational Resources Information Center

Liang, Kun-Hsia; And Others

1995-01-01

A computer-assisted, K-fold cross-validation technique is discussed in the framework of canonical correlation analysis of randomly generated data sets. Analysis results suggest that this technique can effectively reduce the contamination of canonical variates and canonical correlations by sample-specific variance components. (Author/SLD)

Chemical measurement of urine volume

NASA Technical Reports Server (NTRS)

Sauer, R. L.

1978-01-01

Chemical method of measuring volume of urine samples using lithium chloride dilution technique, does not interfere with analysis, is faster, and more accurate than standard volumetric of specific gravity/weight techniques. Adaptation of procedure to urinalysis could prove generally practical for hospital mineral balance and catechoamine determinations.

The accuracy of formol-ether concentration in diagnosing soiltransmitted helminths in elementary school 27 Peusangan in Bireuen

NASA Astrophysics Data System (ADS)

Fitriani, C. L.; Panggabean, M.; Pasaribu, A. P.

2018-03-01

Soil-transmitted helminths (STH) or a group of parasitic nematode worms causing human infection through contact with moist soil may contribute to anemia, nutritional disorders, physical and intellectual growth retardation. School-age children are at high risk of STH infection due to frequent contact with soil. Reliable, sensitive, and practical diagnostic are the test series for detecting STH. This study aimed to assess the sensitivity and specificity of the formol-ether concentration (FEC) in the diagnosis of STH when compared to the Kato-Katz technique. The study was designed at state elementary school 27 Peusangan, Bireuen. The FEC study on a total of 80 (100%) elementary students showed that 12 (15%) sample had the STH infection, while Kato-Katz technique (Gold standard) showed that 31 (38.75%) sample had the STH infection. The FEC technique has the sensitivity of (38.71%), specificity of (100%) and accuracy of (76.25%). The Kato-Katz technique is better than the FEC technique for assessing STH in Bireuen due to mild infection.

Automated Broad-Range Molecular Detection of Bacteria in Clinical Samples

PubMed Central

Hoogewerf, Martine; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

2016-01-01

Molecular detection methods, such as quantitative PCR (qPCR), have found their way into clinical microbiology laboratories for the detection of an array of pathogens. Most routinely used methods, however, are directed at specific species. Thus, anything that is not explicitly searched for will be missed. This greatly limits the flexibility and universal application of these techniques. We investigated the application of a rapid universal bacterial molecular identification method, IS-pro, to routine patient samples received in a clinical microbiology laboratory. IS-pro is a eubacterial technique based on the detection and categorization of 16S-23S rRNA gene interspace regions with lengths that are specific for each microbial species. As this is an open technique, clinicians do not need to decide in advance what to look for. We compared routine culture to IS-pro using 66 samples sent in for routine bacterial diagnostic testing. The samples were obtained from patients with infections in normally sterile sites (without a resident microbiota). The results were identical in 20 (30%) samples, IS-pro detected more bacterial species than culture in 31 (47%) samples, and five of the 10 culture-negative samples were positive with IS-pro. The case histories of the five patients from whom these culture-negative/IS-pro-positive samples were obtained suggest that the IS-pro findings are highly clinically relevant. Our findings indicate that an open molecular approach, such as IS-pro, may have a high added value for clinical practice. PMID:26763956

A LITERATURE REVIEW OF WIPE SAMPLING METHODS FOR CHEMICAL WARFARE AGENTS AND TOXIC INDUSTRIAL CHEMICALS

EPA Science Inventory

Wipe sampling is an important technique for the estimation of contaminant deposition in buildings, homes, or outdoor surfaces as a source of possible human exposure. Numerous

methods of wipe sampling exist, and each method has its own specification for the type of wipe, we...

Scrambled eggs: A highly sensitive molecular diagnostic workflow for Fasciola species specific detection from faecal samples.

PubMed

Calvani, Nichola Eliza Davies; Windsor, Peter Andrew; Bush, Russell David; Šlapeta, Jan

2017-09-01

Fasciolosis, due to Fasciola hepatica and Fasciola gigantica, is a re-emerging zoonotic parasitic disease of worldwide importance. Human and animal infections are commonly diagnosed by the traditional sedimentation and faecal egg-counting technique. However, this technique is time-consuming and prone to sensitivity errors when a large number of samples must be processed or if the operator lacks sufficient experience. Additionally, diagnosis can only be made once the 12-week pre-patent period has passed. Recently, a commercially available coprological antigen ELISA has enabled detection of F. hepatica prior to the completion of the pre-patent period, providing earlier diagnosis and increased throughput, although species differentiation is not possible in areas of parasite sympatry. Real-time PCR offers the combined benefits of highly sensitive species differentiation for medium to large sample sizes. However, no molecular diagnostic workflow currently exists for the identification of Fasciola spp. in faecal samples. A new molecular diagnostic workflow for the highly-sensitive detection and quantification of Fasciola spp. in faecal samples was developed. The technique involves sedimenting and pelleting the samples prior to DNA isolation in order to concentrate the eggs, followed by disruption by bead-beating in a benchtop homogeniser to ensure access to DNA. Although both the new molecular workflow and the traditional sedimentation technique were sensitive and specific, the new molecular workflow enabled faster sample throughput in medium to large epidemiological studies, and provided the additional benefit of speciation. Further, good correlation (R2 = 0.74-0.76) was observed between the real-time PCR values and the faecal egg count (FEC) using the new molecular workflow for all herds and sampling periods. Finally, no effect of storage in 70% ethanol was detected on sedimentation and DNA isolation outcomes; enabling transport of samples from endemic to non-endemic countries without the requirement of a complete cold chain. The commercially-available ELISA displayed poorer sensitivity, even after adjustment of the positive threshold (65-88%), compared to the sensitivity (91-100%) of the new molecular diagnostic workflow. Species-specific assays for sensitive detection of Fasciola spp. enable ante-mortem diagnosis in both human and animal settings. This includes Southeast Asia where there are potentially many undocumented human cases and where post-mortem examination of production animals can be difficult. The new molecular workflow provides a sensitive and quantitative diagnostic approach for the rapid testing of medium to large sample sizes, potentially superseding the traditional sedimentation and FEC technique and enabling surveillance programs in locations where animal and human health funding is limited.

Extraction of citral oil from lemongrass (Cymbopogon Citratus) by steam-water distillation technique

NASA Astrophysics Data System (ADS)

Alam, P. N.; Husin, H.; Asnawi, T. M.; Adisalamun

2018-04-01

In Indonesia, production of citral oil from lemon grass (Cymbopogon Cytratus) is done by a traditional technique whereby a low yield results. To improve the yield, an appropriate extraction technology is required. In this research, a steam-water distillation technique was applied to extract the essential oil from the lemongrass. The effects of sample particle size and bed volume on yield and quality of citral oil produced were investigated. The drying and refining time of 2 hours were used as fixed variables. This research results that minimum citral oil yield of 0.53% was obtained on sample particle size of 3 cm and bed volume of 80%, whereas the maximum yield of 1.95% on sample particle size of 15 cm and bed volume of 40%. The lowest specific gravity of 0.80 and the highest specific gravity of 0.905 were obtained on sample particle size of 8 cm with bed volume of 80% and particle size of 12 cm with bed volume of 70%, respectively. The lowest refractive index of 1.480 and the highest refractive index of 1.495 were obtained on sample particle size of 8 cm with bed volume of 70% and sample particle size of 15 cm with bed volume of 40%, respectively. The solubility of the produced citral oil in alcohol was 70% in ratio of 1:1, and the citral oil concentration obtained was around 79%.

Navigating complex sample analysis using national survey data.

PubMed

Saylor, Jennifer; Friedmann, Erika; Lee, Hyeon Joo

2012-01-01

The National Center for Health Statistics conducts the National Health and Nutrition Examination Survey and other national surveys with probability-based complex sample designs. Goals of national surveys are to provide valid data for the population of the United States. Analyses of data from population surveys present unique challenges in the research process but are valuable avenues to study the health of the United States population. The aim of this study was to demonstrate the importance of using complex data analysis techniques for data obtained with complex multistage sampling design and provide an example of analysis using the SPSS Complex Samples procedure. Illustration of challenges and solutions specific to secondary data analysis of national databases are described using the National Health and Nutrition Examination Survey as the exemplar. Oversampling of small or sensitive groups provides necessary estimates of variability within small groups. Use of weights without complex samples accurately estimates population means and frequency from the sample after accounting for over- or undersampling of specific groups. Weighting alone leads to inappropriate population estimates of variability, because they are computed as if the measures were from the entire population rather than a sample in the data set. The SPSS Complex Samples procedure allows inclusion of all sampling design elements, stratification, clusters, and weights. Use of national data sets allows use of extensive, expensive, and well-documented survey data for exploratory questions but limits analysis to those variables included in the data set. The large sample permits examination of multiple predictors and interactive relationships. Merging data files, availability of data in several waves of surveys, and complex sampling are techniques used to provide a representative sample but present unique challenges. In sophisticated data analysis techniques, use of these data is optimized.

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.

Use of the Genomic Subtractive Hybridization Technique To Develop a Real-Time PCR Assay for Quantitative Detection of Prevotella spp. in Oral Biofilm Samples

PubMed Central

Nagashima, Shiori; Yoshida, Akihiro; Suzuki, Nao; Ansai, Toshihiro; Takehara, Tadamichi

2005-01-01

Genomic subtractive hybridization was used to design Prevotella nigrescens-specific primers and TaqMan probes. Based on this technique, a TaqMan real-time PCR assay was developed for quantifying four oral black-pigmented Prevotella species. The combination of real-time PCR and genomic subtractive hybridization is useful for preparing species-specific primer-probe sets for closely related species. PMID:15956428

Breed traceability of buffalo meat using microsatellite genotyping technique.

PubMed

Kannur, Bheemashankar H; Fairoze, Md Nadeem; Girish, P S; Karabasanavar, Nagappa; Rudresh, B H

2017-02-01

Although buffalo has emerged as a major meat producing animal in Asia, major research on breed traceability has so far been focused on cattle (beef). This research gap on buffalo breed traceability has impelled development and validation of buffalo breed traceability using a set of eight microsatellite (STR) markers in seven Indian buffalo breeds (Bhadawari, Jaffaarabadi, Murrah, Mehsana, Nagpuri, Pandharpuri and Surti). Probability of sharing same profile by two individuals at a specific locus was computed considering different STR numbers, allele pooling in breed and population. Match probabilities per breed were considered and six most polymorphic loci were genotyped. Out of eight microsatellite markers studied, markers CSSMO47, DRB3 and CSSM060 were found most polymorphic. Developed technique was validated with known and unknown, blood and meat samples; wherein, samples were genetically traced in 24 out of 25 samples tested. Results of this study showed potential applications of the methodology and encourage other researchers to address the problem of buffalo traceability so as to create a world-wide archive of breed specific genotypes. This work is the first report of breed traceability of buffalo meat utilizing microsatellite genotyping technique.

Science in Drama: Using Television Programmes to Teach Concepts and Techniques

ERIC Educational Resources Information Center

Rutter, Gordon

2011-01-01

By using a specific episode of the popular television cartoon series "The Simpsons," a range of techniques can be communicated, including microscope setup and use, simple chemical analysis, observation, and interpretation. Knowledge of blood groups and typing, morphological comparison of hair samples, fingerprint analysis, and DNA fingerprinting…

Can groundwater sampling techniques used in monitoring wells influence methane concentrations and isotopes?

PubMed

Rivard, Christine; Bordeleau, Geneviève; Lavoie, Denis; Lefebvre, René; Malet, Xavier

2018-03-06

Methane concentrations and isotopic composition in groundwater are the focus of a growing number of studies. However, concerns are often expressed regarding the integrity of samples, as methane is very volatile and may partially exsolve during sample lifting in the well and transfer to sampling containers. While issues concerning bottle-filling techniques have already been documented, this paper documents a comparison of methane concentration and isotopic composition obtained with three devices commonly used to retrieve water samples from dedicated observation wells. This work lies within the framework of a larger project carried out in the Saint-Édouard area (southern Québec, Canada), whose objective was to assess the risk to shallow groundwater quality related to potential shale gas exploitation. The selected sampling devices, which were tested on ten wells during three sampling campaigns, consist of an impeller pump, a bladder pump, and disposable sampling bags (HydraSleeve). The sampling bags were used both before and after pumping, to verify the appropriateness of a no-purge approach, compared to the low-flow approach involving pumping until stabilization of field physicochemical parameters. Results show that methane concentrations obtained with the selected sampling techniques are usually similar and that there is no systematic bias related to a specific technique. Nonetheless, concentrations can sometimes vary quite significantly (up to 3.5 times) for a given well and sampling event. Methane isotopic composition obtained with all sampling techniques is very similar, except in some cases where sampling bags were used before pumping (no-purge approach), in wells where multiple groundwater sources enter the borehole.

Molecular identification of Coccidioides spp. in soil samples from Brazil

PubMed Central

2011-01-01

Background Since 1991 several outbreaks of acute coccidioidomycosis (CM) were diagnosed in the semi-arid Northeast of Brazil, mainly related to disturbance of armadillo burrows caused by hunters while digging them for the capture of these animals. This activity causes dust contaminated with arthroconidia of Coccidioides posadasii, which, once inhaled, cause the mycosis. We report on the identification of C. posadasii in soil samples related to outbreaks of CM. Results Twenty four soil samples had their DNA extracted and subsequently submitted to a semi-nested PCR technique using specific primers. While only 6 (25%) soil samples were positive for C. posadasii by mice inoculation, all (100%) were positive by the molecular tool. Conclusion This methodology represents a simple, sensitive and specific molecular technique to determine the environmental distribution of Coccidioides spp. in endemic areas, but cannot distinguish the species. Moreover, it may be useful to identify culture isolates. Key-words: 1. Coccidioidomycosis. 2. Coccidioides spp. 3. C. posadasii. 4. Semi-arid. 5. Semi-nested PCR PMID:21575248

Comparison of PIXE and XRF analysis of airborne particulate matter samples collected on Teflon and quartz fibre filters

NASA Astrophysics Data System (ADS)

Chiari, M.; Yubero, E.; Calzolai, G.; Lucarelli, F.; Crespo, J.; Galindo, N.; Nicolás, J. F.; Giannoni, M.; Nava, S.

2018-02-01

Within the framework of research projects focusing on the sampling and analysis of airborne particulate matter, Particle Induced X-ray Emission (PIXE) and Energy Dispersive X-ray Fluorescence (ED-XRF) techniques are routinely used in many laboratories throughout the world to determine the elemental concentration of the particulate matter samples. In this work an inter-laboratory comparison of the results obtained from analysing several samples (collected on both Teflon and quartz fibre filters) using both techniques is presented. The samples were analysed by PIXE (in Florence, at the 3 MV Tandetron accelerator of INFN-LABEC laboratory) and by XRF (in Elche, using the ARL Quant'X EDXRF spectrometer with specific conditions optimized for specific groups of elements). The results from the two sets of measurements are in good agreement for all the analysed samples, thus validating the use of the ARL Quant'X EDXRF spectrometer and the selected measurement protocol for the analysis of aerosol samples. Moreover, thanks to the comparison of PIXE and XRF results on Teflon and quartz fibre filters, possible self-absorption effects due to the penetration of the aerosol particles inside the quartz fibre-filters were quantified.

Development of MRM-based assays for the absolute quantitation of plasma proteins.

PubMed

Kuzyk, Michael A; Parker, Carol E; Domanski, Dominik; Borchers, Christoph H

2013-01-01

Multiple reaction monitoring (MRM), sometimes called selected reaction monitoring (SRM), is a directed tandem mass spectrometric technique performed on to triple quadrupole mass spectrometers. MRM assays can be used to sensitively and specifically quantify proteins based on peptides that are specific to the target protein. Stable-isotope-labeled standard peptide analogues (SIS peptides) of target peptides are added to enzymatic digests of samples, and quantified along with the native peptides during MRM analysis. Monitoring of the intact peptide and a collision-induced fragment of this peptide (an ion pair) can be used to provide information on the absolute peptide concentration of the peptide in the sample and, by inference, the concentration of the intact protein. This technique provides high specificity by selecting for biophysical parameters that are unique to the target peptides: (1) the molecular weight of the peptide, (2) the generation of a specific fragment from the peptide, and (3) the HPLC retention time during LC/MRM-MS analysis. MRM is a highly sensitive technique that has been shown to be capable of detecting attomole levels of target peptides in complex samples such as tryptic digests of human plasma. This chapter provides a detailed description of how to develop and use an MRM protein assay. It includes sections on the critical "first step" of selecting the target peptides, as well as optimization of MRM acquisition parameters for maximum sensitivity of the ion pairs that will be used in the final method, and characterization of the final MRM assay.

High-throughput immunomagnetic scavenging technique for quantitative analysis of live VX nerve agent in water, hamburger, and soil matrixes.

PubMed

Knaack, Jennifer S; Zhou, Yingtao; Abney, Carter W; Prezioso, Samantha M; Magnuson, Matthew; Evans, Ronald; Jakubowski, Edward M; Hardy, Katelyn; Johnson, Rudolph C

2012-11-20

We have developed a novel immunomagnetic scavenging technique for extracting cholinesterase inhibitors from aqueous matrixes using biological targeting and antibody-based extraction. The technique was characterized using the organophosphorus nerve agent VX. The limit of detection for VX in high-performance liquid chromatography (HPLC)-grade water, defined as the lowest calibrator concentration, was 25 pg/mL in a small, 500 μL sample. The method was characterized over the course of 22 sample sets containing calibrators, blanks, and quality control samples. Method precision, expressed as the mean relative standard deviation, was less than 9.2% for all calibrators. Quality control sample accuracy was 102% and 100% of the mean for VX spiked into HPLC-grade water at concentrations of 2.0 and 0.25 ng/mL, respectively. This method successfully was applied to aqueous extracts from soil, hamburger, and finished tap water spiked with VX. Recovery was 65%, 81%, and 100% from these matrixes, respectively. Biologically based extractions of organophosphorus compounds represent a new technique for sample extraction that provides an increase in extraction specificity and sensitivity.

Multiple double cross-section transmission electron microscope sample preparation of specific sub-10 nm diameter Si nanowire devices.

PubMed

Gignac, Lynne M; Mittal, Surbhi; Bangsaruntip, Sarunya; Cohen, Guy M; Sleight, Jeffrey W

2011-12-01

The ability to prepare multiple cross-section transmission electron microscope (XTEM) samples from one XTEM sample of specific sub-10 nm features was demonstrated. Sub-10 nm diameter Si nanowire (NW) devices were initially cross-sectioned using a dual-beam focused ion beam system in a direction running parallel to the device channel. From this XTEM sample, both low- and high-resolution transmission electron microscope (TEM) images were obtained from six separate, specific site Si NW devices. The XTEM sample was then re-sectioned in four separate locations in a direction perpendicular to the device channel: 90° from the original XTEM sample direction. Three of the four XTEM samples were successfully sectioned in the gate region of the device. From these three samples, low- and high-resolution TEM images of the Si NW were taken and measurements of the NW diameters were obtained. This technique demonstrated the ability to obtain high-resolution TEM images in directions 90° from one another of multiple, specific sub-10 nm features that were spaced 1.1 μm apart.

Chi-squared and C statistic minimization for low count per bin data. [sampling in X ray astronomy

NASA Technical Reports Server (NTRS)

Nousek, John A.; Shue, David R.

1989-01-01

Results are presented from a computer simulation comparing two statistical fitting techniques on data samples with large and small counts per bin; the results are then related specifically to X-ray astronomy. The Marquardt and Powell minimization techniques are compared by using both to minimize the chi-squared statistic. In addition, Cash's C statistic is applied, with Powell's method, and it is shown that the C statistic produces better fits in the low-count regime than chi-squared.

A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples.

PubMed

Isaksson, Mats; Hagström, Åsa; Armua-Fernandez, Maria Teresa; Wahlström, Helene; Ågren, Erik Olof; Miller, Andrea; Holmberg, Anders; Lukacs, Morten; Casulli, Adriano; Deplazes, Peter; Juremalm, Mikael

2014-12-19

Following the first finding of Echinococcus multilocularis in Sweden in 2011, 2985 red foxes (Vulpes vulpes) were analysed by the segmental sedimentation and counting technique. This is a labour intensive method and requires handling of the whole carcass of the fox, resulting in a costly analysis. In an effort to reduce the cost of labour and sample handling, an alternative method has been developed. The method is sensitive and partially automated for detection of E. multilocularis in faecal samples. The method has been used in the Swedish E. multilocularis monitoring program for 2012-2013 on more than 2000 faecal samples. We describe a new semi-automated magnetic capture probe DNA extraction method and real time hydrolysis probe polymerase chain reaction assay (MC-PCR) for the detection of E. multilocularis DNA in faecal samples from red fox. The diagnostic sensitivity was determined by validating the new method against the sedimentation and counting technique in fox samples collected in Switzerland where E. multilocularis is highly endemic. Of 177 foxes analysed by the sedimentation and counting technique, E. multilocularis was detected in 93 animals. Eighty-two (88%, 95% C.I 79.8-93.9) of these were positive in the MC-PCR. In foxes with more than 100 worms, the MC-PCR was positive in 44 out of 46 (95.7%) cases. The two MC-PCR negative samples originated from foxes with only immature E. multilocularis worms. In foxes with 100 worms or less, (n = 47), 38 (80.9%) were positive in the MC-PCR. The diagnostic specificity of the MC-PCR was evaluated using fox scats collected within the Swedish screening. Of 2158 samples analysed, two were positive. This implies that the specificity is at least 99.9% (C.I. = 99.7-100). The MC-PCR proved to have a high sensitivity and a very high specificity. The test is partially automated but also possible to perform manually if desired. The test is well suited for nationwide E. multilocularis surveillance programs where sampling of fox scats is done to reduce the costs for sampling and where a test with a high sensitivity and a very high specificity is needed.

Isoschizomers and amplified fragment length polymorphism for the detection of specific cytosine methylation changes.

PubMed

Ruiz-García, Leonor; Cabezas, Jose Antonio; de María, Nuria; Cervera, María-Teresa

2010-01-01

Different molecular techniques have been developed to study either the global level of methylated cytosines or methylation at specific gene sequences. One of them is a modification of the Amplified Fragment Length Polymorphism (AFLP) technique that has been used to study methylation of anonymous CCGG sequences in different fungi, plant and animal species. The main variation of this technique is based on the use of isoschizomers with different methylation sensitivity (such as HpaII and MspI) as a frequent cutter restriction enzyme. For each sample, AFLP analysis is performed using both EcoRI/HpaII and EcoRI/MspI digested samples. Comparative analysis between EcoRI/HpaII and EcoRI/MspI fragment patterns allows the identification of two types of polymorphisms: (1) "Methylation-insensitive polymorphisms" that show common EcoRI/HpaII and EcoRI/MspI patterns but are detected as polymorphic amplified fragments among samples; and (2) "Methylation-sensitive polymorphisms" that are associated with amplified fragments differing in their presence or absence or in their intensity between EcoRI/HpaII and EcoRI/MspI patterns. This chapter describes a detailed protocol of this technique and discusses modifications that can be applied to adjust the technology to different species of interest.

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.

Evaluation of two-dimensional electrophoresis and liquid chromatography – tandem mass spectrometry for tissue-specific protein profiling of laser-microdissected plant samples

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

Schad, Martina; Lipton, Mary S.; Giavalisco, Patrick

2005-07-14

Laser microdissection (LM) allows the collection of homogeneous tissue- and cell specific plant samples. The employment of this technique with subsequent protein analysis has thus far not been reported for plant tissues, probably due to the difficulties associated with defining a reasonable cellular morphology and, in parallel, allowing efficient protein extraction from tissue samples. The relatively large sample amount needed for successful proteome analysis is an additional issue that complicates protein profiling on a tissue- or even cell-specific level. In contrast to transcript profiling that can be performed from very small sample amounts due to efficient amplification strategies, there ismore » as yet no amplification procedure for proteins available. In the current study, we compared different tissue preparation techniques prior to LM/laser pressure catapulting (LMPC) with respect to their suitability for protein retrieval. Cryosectioning was identified as the best compromise between tissue morphology and effective protein extraction. After collection of vascular bundles from Arabidopsis thaliana stem tissue by LMPC, proteins were extracted and subjected to protein analysis, either by classical two-dimensional gel electrophoresis (2-DE), or by high-efficiency liquid chromatography (LC) in conjunction with tandem mass spectrometry (MS/MS). Our results demonstrate that both methods can be used with LMPC collected plant material. But because of the significantly lower sample amount required for LC-MS/MS than for 2-DE, the combination of LMPC and LC-MS/MS has a higher potential to promote comprehensive proteome analysis of specific plant tissues.« less

Recent developments and future trends in solid phase microextraction techniques towards green analytical chemistry.

PubMed

Spietelun, Agata; Marcinkowski, Łukasz; de la Guardia, Miguel; Namieśnik, Jacek

2013-12-20

Solid phase microextraction find increasing applications in the sample preparation step before chromatographic determination of analytes in samples with a complex composition. These techniques allow for integrating several operations, such as sample collection, extraction, analyte enrichment above the detection limit of a given measuring instrument and the isolation of analytes from sample matrix. In this work the information about novel methodological and instrumental solutions in relation to different variants of solid phase extraction techniques, solid-phase microextraction (SPME), stir bar sorptive extraction (SBSE) and magnetic solid phase extraction (MSPE) is presented, including practical applications of these techniques and a critical discussion about their advantages and disadvantages. The proposed solutions fulfill the requirements resulting from the concept of sustainable development, and specifically from the implementation of green chemistry principles in analytical laboratories. Therefore, particular attention was paid to the description of possible uses of novel, selective stationary phases in extraction techniques, inter alia, polymeric ionic liquids, carbon nanotubes, and silica- and carbon-based sorbents. The methodological solutions, together with properly matched sampling devices for collecting analytes from samples with varying matrix composition, enable us to reduce the number of errors during the sample preparation prior to chromatographic analysis as well as to limit the negative impact of this analytical step on the natural environment and the health of laboratory employees. Copyright © 2013 Elsevier B.V. All rights reserved.

Schnellverfahren zur flammenlosen AAS-Bestimmung von Spurenelementen in geologischen Proben

NASA Astrophysics Data System (ADS)

Schrön, W.; Bombach, G.; Beuge, P.

This paper reports experience with direct quantitative trace element determinations in powdered geological samples by nameless atomic absorption spectroscopy. Two methods were explored. The first one is based on the production of a sample aerosol by laser radiation in a specifically designed sample chamber and the subsequent transport of the aerosol into a graphite tube, which has been preheated to a stable temperature. This technique is suited for a large range of concentration and is relatively free from matrix interferences. The technique was tested for the elements Ag, As, Bi, Cd, Co, Mn, Ni, Pb, Sb, Se, Sr and Tl. The described sample chamber can be also used in combination with other spcctroscopic techniques. The second method explored permits the quantitative determination of trace elements at very low concentrations. Essentially an accurately weighed amount of sample is placed on a graphite rod and introduced into a graphite furnace by inserting the rod through the sample injection port. Atomization takes place also under stable temperature conditions. Using this technique detection limits were found to be 10 -11 g for Ag, 2 × 10 -11 g for Cd and 10 -10 g for Sb in silicate materials.

Effect of different mixing methods on the physical properties of Portland cement.

PubMed

Shahi, Shahriar; Ghasemi, Negin; Rahimi, Saeed; Yavari, Hamidreza; Samiei, Mohammad; Jafari, Farnaz

2016-12-01

The Portland cement is hydrophilic cement; as a result, the powder-to-liquid ratio affects the properties of the final mix. In addition, the mixing technique affects hydration. The aim of this study was to evaluate the effect of different mixing techniques (conventional, amalgamator and ultrasonic) on some selective physical properties of Portland cement. The physical properties to be evaluated were determined using the ISO 6786:2001 specification. One hundred sixty two samples of Portland cement were prepared for three mixing techniques for each physical property (each 6 samples). Data were analyzed using descriptive statistics, one-way ANOVA and post hoc Tukey tests. Statistical significance was set at P <0.05. The mixing technique had no significant effect on the compressive strength, film thickness and flow of Portland cement ( P >0.05). Dimensional changes (shrinkage), solubility and pH increased significantly by amalgamator and ultrasonic mixing techniques ( P <0.05). The ultrasonic technique significantly decreased working time, and the amalgamator and ultrasonic techniques significantly decreased the setting time ( P <0.05). The mixing technique exerted no significant effect on the flow, film thickness and compressive strength of Portland cement samples. Key words: Physical properties, Portland cement, mixing methods.

Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems.

PubMed

Ximenes, Camila; Brandão, Eduardo; Oliveira, Paula; Rocha, Abraham; Rego, Tamisa; Medeiros, Rafael; Aguiar-Santos, Ana; Ferraz, João; Reis, Christian; Araujo, Paulo; Carvalho, Luiz; Melo, Fabio L

2014-12-01

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

An Exploratory Study of Animal-Assisted Interventions Utilized by Mental Health Professionals

ERIC Educational Resources Information Center

O'Callaghan, Dana M.; Chandler, Cynthia K.

2011-01-01

This study implemented an exploratory analysis to examine how a sample of mental health professionals incorporates specific animal-assisted techniques into the therapeutic process. An extensive review of literature related to animal-assisted therapy (AAT) resulted in the identification of 18 techniques and 10 intentions for the practice of AAT in…

Air pollution source identification

NASA Technical Reports Server (NTRS)

Fordyce, J. S.

1975-01-01

Techniques for air pollution source identification are reviewed, and some results obtained with them are evaluated. Described techniques include remote sensing from satellites and aircraft, on-site monitoring, and the use of injected tracers and pollutants themselves as tracers. The use of a large number of trace elements in ambient airborne particulate matter as a practical means of identifying sources is discussed in detail. Sampling and analysis techniques are described, and it is shown that elemental constituents can be related to specific source types such as those found in the earth's crust and those associated with specific industries. Source identification sytems are noted which utilize charged particle X-ray fluorescence analysis of original field data.

Effect of chromium substitution on the dielectric properties of mixed Ni-Zn ferrite prepared by WOWS sol–gel technique

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

Ashtar, M.; Munir, A.; Anis-ur-Rehman, M.

2016-07-15

Graphical abstract: Variation of AC conductivity (σ{sub AC}) as a function of natural log of angular frequency (lnω) for Ni{sub 0.5}Zn{sub 0.5}Fe{sub 2-x}Cr{sub x}O{sub 4} nanoferrites at room temperature. - Highlights: • Cr doped mixed Ni-Zn ferrites were successfully synthesized by a newly developed WOWS sol gel technique. • The specific surface area and specific surface area to volume ratio increased with decrease in particle size. • The resonance peaks appeared in dielectric loss graphs, shifting towards low frequency with the increase in Cr concentration. • The prepared samples have the lowest values of the dielectric constant. • The dielectricmore » constant were observed to be inversely proportional to square root of the AC resistivity. - Abstract: Cr{sup +3} doped Ni-Zn nanoferrite samples with composition Ni{sub 0.5}Zn{sub 0.5}Fe{sub 2-x}Cr{sub x}O{sub 4}(x = 0.1, 0.2, 0.3, 0.4) were synthesized With Out Water and Surfactant (WOWS) sol-gel technique. The structural, morphological and dielectric properties of the samples were investigated. The lattice constant, crystallite size, theoretical density and porosity of each sample were obtained from X-ray diffraction (XRD) data. The specific surface area and specific surface area to volume ratio increased with the decrease in the size of Cr{sup +3} doped Ni-Zn ferrite nanoparticles, as the concentration of Cr{sup +3} increased. The SEM analysis revealed that the particles were of nano size and of spherical shape. The dielectric parameters such as dielectric constant (ε′) and dielectric loss (tanδ) of all the samples as a function of frequency at room temperature were measured. The AC conductivity (σ{sub AC}) was determined from the dielectric parameters, which showed increasing trend with the rise in frequency.« less

Scrambled eggs: A highly sensitive molecular diagnostic workflow for Fasciola species specific detection from faecal samples

PubMed Central

Calvani, Nichola Eliza Davies; Windsor, Peter Andrew; Bush, Russell David

2017-01-01

Background Fasciolosis, due to Fasciola hepatica and Fasciola gigantica, is a re-emerging zoonotic parasitic disease of worldwide importance. Human and animal infections are commonly diagnosed by the traditional sedimentation and faecal egg-counting technique. However, this technique is time-consuming and prone to sensitivity errors when a large number of samples must be processed or if the operator lacks sufficient experience. Additionally, diagnosis can only be made once the 12-week pre-patent period has passed. Recently, a commercially available coprological antigen ELISA has enabled detection of F. hepatica prior to the completion of the pre-patent period, providing earlier diagnosis and increased throughput, although species differentiation is not possible in areas of parasite sympatry. Real-time PCR offers the combined benefits of highly sensitive species differentiation for medium to large sample sizes. However, no molecular diagnostic workflow currently exists for the identification of Fasciola spp. in faecal samples. Methodology/Principal findings A new molecular diagnostic workflow for the highly-sensitive detection and quantification of Fasciola spp. in faecal samples was developed. The technique involves sedimenting and pelleting the samples prior to DNA isolation in order to concentrate the eggs, followed by disruption by bead-beating in a benchtop homogeniser to ensure access to DNA. Although both the new molecular workflow and the traditional sedimentation technique were sensitive and specific, the new molecular workflow enabled faster sample throughput in medium to large epidemiological studies, and provided the additional benefit of speciation. Further, good correlation (R2 = 0.74–0.76) was observed between the real-time PCR values and the faecal egg count (FEC) using the new molecular workflow for all herds and sampling periods. Finally, no effect of storage in 70% ethanol was detected on sedimentation and DNA isolation outcomes; enabling transport of samples from endemic to non-endemic countries without the requirement of a complete cold chain. The commercially-available ELISA displayed poorer sensitivity, even after adjustment of the positive threshold (65–88%), compared to the sensitivity (91–100%) of the new molecular diagnostic workflow. Conclusions/Significance Species-specific assays for sensitive detection of Fasciola spp. enable ante-mortem diagnosis in both human and animal settings. This includes Southeast Asia where there are potentially many undocumented human cases and where post-mortem examination of production animals can be difficult. The new molecular workflow provides a sensitive and quantitative diagnostic approach for the rapid testing of medium to large sample sizes, potentially superseding the traditional sedimentation and FEC technique and enabling surveillance programs in locations where animal and human health funding is limited. PMID:28915255

Multiplex Amplification Refractory Mutation System PCR (ARMS-PCR) provides sequencing independent typing of canine parvovirus.

PubMed

Chander, Vishal; Chakravarti, Soumendu; Gupta, Vikas; Nandi, Sukdeb; Singh, Mithilesh; Badasara, Surendra Kumar; Sharma, Chhavi; Mittal, Mitesh; Dandapat, S; Gupta, V K

2016-12-01

Canine parvovirus-2 antigenic variants (CPV-2a, CPV-2b and CPV-2c) ubiquitously distributed worldwide in canine population causes severe fatal gastroenteritis. Antigenic typing of CPV-2 remains a prime focus of research groups worldwide in understanding the disease epidemiology and virus evolution. The present study was thus envisioned to provide a simple sequencing independent, rapid, robust, specific, user-friendly technique for detecting and typing of presently circulating CPV-2 antigenic variants. ARMS-PCR strategy was employed using specific primers for CPV-2a, CPV-2b and CPV-2c to differentiate these antigenic types. ARMS-PCR was initially optimized with reference positive controls in two steps; where first reaction was used to differentiate CPV-2a from CPV-2b/CPV-2c. The second reaction was carried out with CPV-2c specific primers to confirm the presence of CPV-2c. Initial validation of the ARMS-PCR was carried out with 24 sequenced samples and the results were matched with the sequencing results. ARMS-PCR technique was further used to screen and type 90 suspected clinical samples. Randomly selected 15 suspected clinical samples that were typed with this technique were sequenced. The results of ARMS-PCR and the sequencing matched exactly with each other. The developed technique has a potential to become a sequencing independent method for simultaneous detection and typing of CPV-2 antigenic variants in veterinary disease diagnostic laboratories globally. Copyright © 2016 Elsevier B.V. All rights reserved.

Mars chronology: Assessing techniques for quantifying surficial processes

USGS Publications Warehouse

Doran, P.T.; Clifford, S.M.; Forman, S.L.; Nyquist, Larry; Papanastassiou, D.A.; Stewart, B.W.; Sturchio, N.C.; Swindle, T.D.; Cerling, T.; Kargel, J.; McDonald, G.; Nishiizumi, K.; Poreda, R.; Rice, J.W.; Tanaka, K.

2004-01-01

Currently, the absolute chronology of Martian rocks, deposits and events is based mainly on crater counting and remains highly imprecise with epoch boundary uncertainties in excess of 2 billion years. Answers to key questions concerning the comparative origin and evolution of Mars and Earth will not be forthcoming without a rigid Martian chronology, enabling the construction of a time scale comparable to Earth's. Priorities for exploration include calibration of the cratering rate, dating major volcanic and fluvial events and establishing chronology of the polar layered deposits. If extinct and/or extant life is discovered, the chronology of the biosphere will be of paramount importance. Many radiometric and cosmogenic techniques applicable on Earth and the Moon will apply to Mars after certain baselines (e.g. composition of the atmosphere, trace species, chemical and physical characteristics of Martian dust) are established. The high radiation regime may pose a problem for dosimetry-based techniques (e.g. luminescence). The unique isotopic composition of nitrogen in the Martian atmosphere may permit a Mars-specific chronometer for tracing the time-evolution of the atmosphere and of lithic phases with trapped atmospheric gases. Other Mars-specific chronometers include measurement of gas fluxes and accumulation of platinum group elements (PGE) in the regolith. Putting collected samples into geologic context is deemed essential, as is using multiple techniques on multiple samples. If in situ measurements are restricted to a single technique it must be shown to give consistent results on multiple samples, but in all cases, using two or more techniques (e.g. on the same lander) will reduce error. While there is no question that returned samples will yield the best ages, in situ techniques have the potential to be flown on multiple missions providing a larger data set and broader context in which to place the more accurate dates. ?? 2004 Elsevier B.V. All rights reserved.

Improving EEG-Based Motor Imagery Classification for Real-Time Applications Using the QSA Method.

PubMed

Batres-Mendoza, Patricia; Ibarra-Manzano, Mario A; Guerra-Hernandez, Erick I; Almanza-Ojeda, Dora L; Montoro-Sanjose, Carlos R; Romero-Troncoso, Rene J; Rostro-Gonzalez, Horacio

2017-01-01

We present an improvement to the quaternion-based signal analysis (QSA) technique to extract electroencephalography (EEG) signal features with a view to developing real-time applications, particularly in motor imagery (IM) cognitive processes. The proposed methodology (iQSA, improved QSA) extracts features such as the average, variance, homogeneity, and contrast of EEG signals related to motor imagery in a more efficient manner (i.e., by reducing the number of samples needed to classify the signal and improving the classification percentage) compared to the original QSA technique. Specifically, we can sample the signal in variable time periods (from 0.5 s to 3 s, in half-a-second intervals) to determine the relationship between the number of samples and their effectiveness in classifying signals. In addition, to strengthen the classification process a number of boosting-technique-based decision trees were implemented. The results show an 82.30% accuracy rate for 0.5 s samples and 73.16% for 3 s samples. This is a significant improvement compared to the original QSA technique that offered results from 33.31% to 40.82% without sampling window and from 33.44% to 41.07% with sampling window, respectively. We can thus conclude that iQSA is better suited to develop real-time applications.

Improving EEG-Based Motor Imagery Classification for Real-Time Applications Using the QSA Method

PubMed Central

Batres-Mendoza, Patricia; Guerra-Hernandez, Erick I.; Almanza-Ojeda, Dora L.; Montoro-Sanjose, Carlos R.

2017-01-01

We present an improvement to the quaternion-based signal analysis (QSA) technique to extract electroencephalography (EEG) signal features with a view to developing real-time applications, particularly in motor imagery (IM) cognitive processes. The proposed methodology (iQSA, improved QSA) extracts features such as the average, variance, homogeneity, and contrast of EEG signals related to motor imagery in a more efficient manner (i.e., by reducing the number of samples needed to classify the signal and improving the classification percentage) compared to the original QSA technique. Specifically, we can sample the signal in variable time periods (from 0.5 s to 3 s, in half-a-second intervals) to determine the relationship between the number of samples and their effectiveness in classifying signals. In addition, to strengthen the classification process a number of boosting-technique-based decision trees were implemented. The results show an 82.30% accuracy rate for 0.5 s samples and 73.16% for 3 s samples. This is a significant improvement compared to the original QSA technique that offered results from 33.31% to 40.82% without sampling window and from 33.44% to 41.07% with sampling window, respectively. We can thus conclude that iQSA is better suited to develop real-time applications. PMID:29348744

Arthropod Surveillance Programs: Basic Components, Strategies, and Analysis.

PubMed

Cohnstaedt, Lee W; Rochon, Kateryn; Duehl, Adrian J; Anderson, John F; Barrera, Roberto; Su, Nan-Yao; Gerry, Alec C; Obenauer, Peter J; Campbell, James F; Lysyk, Tim J; Allan, Sandra A

2012-03-01

Effective entomological surveillance planning stresses a careful consideration of methodology, trapping technologies, and analysis techniques. Herein, the basic principles and technological components of arthropod surveillance plans are described, as promoted in the symposium "Advancements in arthropod monitoring technology, techniques, and analysis" presented at the 58th annual meeting of the Entomological Society of America in San Diego, CA. Interdisciplinary examples of arthropod monitoring for urban, medical, and veterinary applications are reviewed. Arthropod surveillance consists of the three components: 1) sampling method, 2) trap technology, and 3) analysis technique. A sampling method consists of selecting the best device or collection technique for a specific location and sampling at the proper spatial distribution, optimal duration, and frequency to achieve the surveillance objective. Optimized sampling methods are discussed for several mosquito species (Diptera: Culicidae) and ticks (Acari: Ixodidae). The advantages and limitations of novel terrestrial and aerial insect traps, artificial pheromones and kairomones are presented for the capture of red flour beetle (Coleoptera: Tenebrionidae), small hive beetle (Coleoptera: Nitidulidae), bed bugs (Hemiptera: Cimicidae), and Culicoides (Diptera: Ceratopogonidae) respectively. After sampling, extrapolating real world population numbers from trap capture data are possible with the appropriate analysis techniques. Examples of this extrapolation and action thresholds are given for termites (Isoptera: Rhinotermitidae) and red flour beetles.

Arthropod Surveillance Programs: Basic Components, Strategies, and Analysis

PubMed Central

Rochon, Kateryn; Duehl, Adrian J.; Anderson, John F.; Barrera, Roberto; Su, Nan-Yao; Gerry, Alec C.; Obenauer, Peter J.; Campbell, James F.; Lysyk, Tim J.; Allan, Sandra A.

2015-01-01

Effective entomological surveillance planning stresses a careful consideration of methodology, trapping technologies, and analysis techniques. Herein, the basic principles and technological components of arthropod surveillance plans are described, as promoted in the symposium “Advancements in arthropod monitoring technology, techniques, and analysis” presented at the 58th annual meeting of the Entomological Society of America in San Diego, CA. Interdisciplinary examples of arthropod monitoring for urban, medical, and veterinary applications are reviewed. Arthropod surveillance consists of the three components: 1) sampling method, 2) trap technology, and 3) analysis technique. A sampling method consists of selecting the best device or collection technique for a specific location and sampling at the proper spatial distribution, optimal duration, and frequency to achieve the surveillance objective. Optimized sampling methods are discussed for several mosquito species (Diptera: Culicidae) and ticks (Acari: Ixodidae). The advantages and limitations of novel terrestrial and aerial insect traps, artificial pheromones and kairomones are presented for the capture of red flour beetle (Coleoptera: Tenebrionidae), small hive beetle (Coleoptera: Nitidulidae), bed bugs (Hemiptera: Cimicidae), and Culicoides (Diptera: Ceratopogonidae) respectively. After sampling, extrapolating real world population numbers from trap capture data are possible with the appropriate analysis techniques. Examples of this extrapolation and action thresholds are given for termites (Isoptera: Rhinotermitidae) and red flour beetles. PMID:26543242

Approaches to Recruiting 'Hard-To-Reach' Populations into Re-search: A Review of the Literature.

PubMed

Shaghaghi, Abdolreza; Bhopal, Raj S; Sheikh, Aziz

2011-01-01

'Hard-to-reach' is a term used to describe those sub-groups of the population that may be difficult to reach or involve in research or public health programmes. Application of a single term to call these sub-sections of populations implies a homogeneity within distinct groups, which does not necessarily exist. Different sampling techniques were introduced so far to recruit hard-to-reach populations. In this article, we have reviewed a range of ap-proaches that have been used to widen participation in studies. We performed a Pubmed and Google search for relevant English language articles using the keywords and phrases: (hard-to-reach AND population* OR sampl*), (hidden AND population* OR sample*) and ("hard to reach" AND population* OR sample*) and a consul-tation of the retrieved articles' bibliographies to extract empirical evidence from publications that discussed or examined the use of sampling techniques to recruit hidden or hard-to-reach populations in health studies. Reviewing the literature has identified a range of techniques to recruit hard-to-reach populations, including snowball sampling, respondent-driven sampling (RDS), indigenous field worker sampling (IFWS), facility-based sampling (FBS), targeted sampling (TS), time-location (space) sampling (TLS), conventional cluster sampling (CCS) and capture re-capture sampling (CR). The degree of compliance with a study by a certain 'hard-to-reach' group de-pends on the characteristics of that group, recruitment technique used and the subject of inter-est. Irrespective of potential advantages or limitations of the recruitment techniques reviewed, their successful use depends mainly upon our knowledge about specific characteristics of the target populations. Thus in line with attempts to expand the current boundaries of our know-ledge about recruitment techniques in health studies and their applications in varying situa-tions, we should also focus on possibly all contributing factors which may have an impact on participation rate within a defined population group.

Isolating RNA from precursor and mature melanocytes from human vitiligo and normal skin using laser capture microdissection.

PubMed

Goldstein, Nathaniel B; Koster, Maranke I; Hoaglin, Laura G; Wright, Michael J; Robinson, Steven E; Robinson, William A; Roop, Dennis R; Norris, David A; Birlea, Stanca A

2016-10-01

To characterize the gene expression profile of regenerated melanocytes in the narrow band UVB (NBUVB)-treated vitiligo epidermis and their precursors in the hair follicle, we present here a strategy of RNA isolation from in situ melanocytes using human frozen skin. We developed a rapid immunostaining protocol using the NKI-beteb antibody, which labels differentiated and precursor melanocytes, followed by fluorescent laser capture microdissection. This technique enabled the direct isolation, from melanocyte and adjacent keratinocyte populations, of satisfactory quality RNA that was successfully amplified and analysed by qRT-PCR. The melanocyte-specific gene transcripts TYR, DCT, TYRP1 and PMEL were significantly upregulated in our NBUVB-treated melanocyte samples as compared with the keratinocyte samples, while keratinocyte-specific genes (KRT5 and KRT14) were expressed significantly higher in the keratinocyte samples as compared with the melanocyte samples. Furthermore, in both NBUVB-treated vitiligo skin and normal skin, when bulge melanocytes were compared with epidermal melanocytes, we found significantly lower expression of melanocyte-specific genes and significantly higher expression of three melanocytic stem cell genes (SOX9, WIF1 and SFRP1), while ALCAM and ALDH1A1 transcripts did not show significant variation. We found significantly higher expression of melanocyte-specific genes in the epidermis of NBUVB-treated vitiligo, as compared to the normal skin. When comparing bulge melanocyte samples from untreated vitiligo, NBUVB-treated vitiligo and normal skin, we did not find significant differences in the expression of melanocyte-specific genes or melanocytic stem cell genes. These techniques offer valuable opportunities to study melanocytes and their precursors in vitiligo and other pigmentation disorders. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Immunofluorescence Technique Using HeLa Cells Expressing Recombinant Nucleoprotein for Detection of Immunoglobulin G Antibodies to Crimean-Congo Hemorrhagic Fever Virus

PubMed Central

Saijo, Masayuki; Qing, Tang; Niikura, Masahiro; Maeda, Akihiko; Ikegami, Tetsuro; Sakai, Koji; Prehaud, Christophe; Kurane, Ichiro; Morikawa, Shigeru

2002-01-01

A HeLa cell line continuously expressing recombinant nucleoprotein (rNP) of the Crimean-Congo hemorrhagic fever virus (CCHFV) was established by transfection with an expression vector containing the cDNA of CCHFV NP (pKS336-CCHFV-NP). These cells were used as antigens for indirect immunofluorescence (IF) to detect immunoglobulin G antibodies to CCHFV. The sensitivity and specificity of this IF technique were examined by using serum samples and were compared to those of the IF technique using CCHFV-infected Vero E6 cells (authentic antigen). Staining of the CCHFV rNP expressed in HeLa cells showed a unique granular pattern similar to that of CCHFV-infected Vero E6 cells. Positive staining could easily be distinguished from a negative result. All 13 serum samples determined to be positive by using the authentic antigen were also determined to be positive by using CCHFV rNP-expressing HeLa cells (recombinant antigen). The 108 serum samples determined to be negative by using the authentic antigen were also determined to be negative by using the recombinant antigen. Thus, both the sensitivity and the specificity of this IF technique were 100% compared to the IF with authentic antigen. The novel IF technique using CCHFV rNP-expressing HeLa cells can be used not only for diagnosis of CCHF but also for epidemiological studies on CCHFV infections. PMID:11825944

Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

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

Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.

Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently,more » the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.« less

Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

DOE PAGES

Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.; ...

2016-05-04

Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently,more » the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.« less

Systematic comparison of static and dynamic headspace sampling techniques for gas chromatography.

PubMed

Kremser, Andreas; Jochmann, Maik A; Schmidt, Torsten C

2016-09-01

Six automated, headspace-based sample preparation techniques were used to extract volatile analytes from water with the goal of establishing a systematic comparison between commonly available instrumental alternatives. To that end, these six techniques were used in conjunction with the same gas chromatography instrument for analysis of a common set of volatile organic carbon (VOC) analytes. The methods were thereby divided into three classes: static sampling (by syringe or loop), static enrichment (SPME and PAL SPME Arrow), and dynamic enrichment (ITEX and trap sampling). For PAL SPME Arrow, different sorption phase materials were also included in the evaluation. To enable an effective comparison, method detection limits (MDLs), relative standard deviations (RSDs), and extraction yields were determined and are discussed for all techniques. While static sampling techniques exhibited sufficient extraction yields (approx. 10-20 %) to be reliably used down to approx. 100 ng L(-1), enrichment techniques displayed extraction yields of up to 80 %, resulting in MDLs down to the picogram per liter range. RSDs for all techniques were below 27 %. The choice on one of the different instrumental modes of operation (aforementioned classes) was thereby the most influential parameter in terms of extraction yields and MDLs. Individual methods inside each class showed smaller deviations, and the least influences were observed when evaluating different sorption phase materials for the individual enrichment techniques. The option of selecting specialized sorption phase materials may, however, be more important when analyzing analytes with different properties such as high polarity or the capability of specific molecular interactions. Graphical Abstract PAL SPME Arrow during the extraction of volatile analytes from the headspace of an aqueous sample.

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping.

PubMed

Roth, Gary A; Sosa Peña, Maria del Pilar; Neu-Baker, Nicole M; Tahiliani, Sahil; Brenner, Sara A

2015-12-08

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the identification, localization, and characterization of nanoscale materials in biological samples is on the rise. Currently available methods, such as electron microscopy, tend to be resource-intensive, making their use prohibitive for much of the research community. Enhanced darkfield microscopy complemented with a hyperspectral imaging system may provide a solution to this bottleneck by enabling rapid and less expensive characterization of nanoparticles in histological samples. This method allows for high-contrast nanoscale imaging as well as nanomaterial identification. For this technique, histological tissue samples are prepared as they would be for light-based microscopy. First, positive control samples are analyzed to generate the reference spectra that will enable the detection of a material of interest in the sample. Negative controls without the material of interest are also analyzed in order to improve specificity (reduce false positives). Samples can then be imaged and analyzed using methods and software for hyperspectral microscopy or matched against these reference spectra in order to provide maps of the location of materials of interest in a sample. The technique is particularly well-suited for materials with highly unique reflectance spectra, such as noble metals, but is also applicable to other materials, such as semi-metallic oxides. This technique provides information that is difficult to acquire from histological samples without the use of electron microscopy techniques, which may provide higher sensitivity and resolution, but are vastly more resource-intensive and time-consuming than light microscopy.

TOF-SIMS imaging technique with information entropy

NASA Astrophysics Data System (ADS)

Aoyagi, Satoka; Kawashima, Y.; Kudo, Masahiro

2005-05-01

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is capable of chemical imaging of proteins on insulated samples in principal. However, selection of specific peaks related to a particular protein, which are necessary for chemical imaging, out of numerous candidates had been difficult without an appropriate spectrum analysis technique. Therefore multivariate analysis techniques, such as principal component analysis (PCA), and analysis with mutual information defined by information theory, have been applied to interpret SIMS spectra of protein samples. In this study mutual information was applied to select specific peaks related to proteins in order to obtain chemical images. Proteins on insulated materials were measured with TOF-SIMS and then SIMS spectra were analyzed by means of the analysis method based on the comparison using mutual information. Chemical mapping of each protein was obtained using specific peaks related to each protein selected based on values of mutual information. The results of TOF-SIMS images of proteins on the materials provide some useful information on properties of protein adsorption, optimality of immobilization processes and reaction between proteins. Thus chemical images of proteins by TOF-SIMS contribute to understand interactions between material surfaces and proteins and to develop sophisticated biomaterials.

Synchronous Teaching Techniques from the Perspective and Observation of Virtual High School Teachers: An Investigative Study

ERIC Educational Resources Information Center

O'Brien, Amy; Fuller, Richard

2018-01-01

This article describes how there are specific tools and techniques used by teachers in synchronous-live-virtual classroom environments. Those items were investigated in this article. Data was collected from semi-structured, one-on-one interviews conducted with a purposeful sample of twelve identified "highly-qualified" teachers at a…

Development of a Tunnel Diode Resonator technique for magnetic measurements in Electrostatic Levitation chamber

NASA Astrophysics Data System (ADS)

Spyrison, N. S.; Prommapan, P.; Kim, H.; Maloney, J.; Rustan, G. E.; Kreyssig, A.; Goldman, A. I.; Prozorov, R.

2011-03-01

The incorporation of the Tunnel Diode Resonator (TDR) technique into an ElectroStatic Levitation (ESL) apparatus was explored. The TDR technique is known to operate and behave well at low temperatures with careful attention to coil-sample positioning in a dark, shielded environment. With these specifications a frequency resolution of 10-9 in a few seconds counting time can be achieved. Complications arise when this technique is applied in the ESL chamber where a sample of molten metal is levitating less then 10 mm from the coil in a large electrostatic field. We have tested a variety of coils unconventional to TDR; including Helmholtz pairs and Archimedean spiral coils. Work was supported by the Nation Science Foundation under grant DMR-08-17157

Combined X-ray CT and mass spectrometry for biomedical imaging applications

NASA Astrophysics Data System (ADS)

Schioppa, E., Jr.; Ellis, S.; Bruinen, A. L.; Visser, J.; Heeren, R. M. A.; Uher, J.; Koffeman, E.

2014-04-01

Imaging technologies play a key role in many branches of science, especially in biology and medicine. They provide an invaluable insight into both internal structure and processes within a broad range of samples. There are many techniques that allow one to obtain images of an object. Different techniques are based on the analysis of a particular sample property by means of a dedicated imaging system, and as such, each imaging modality provides the researcher with different information. The use of multimodal imaging (imaging with several different techniques) can provide additional and complementary information that is not possible when employing a single imaging technique alone. In this study, we present for the first time a multi-modal imaging technique where X-ray computerized tomography (CT) is combined with mass spectrometry imaging (MSI). While X-ray CT provides 3-dimensional information regarding the internal structure of the sample based on X-ray absorption coefficients, MSI of thin sections acquired from the same sample allows the spatial distribution of many elements/molecules, each distinguished by its unique mass-to-charge ratio (m/z), to be determined within a single measurement and with a spatial resolution as low as 1 μm or even less. The aim of the work is to demonstrate how molecular information from MSI can be spatially correlated with 3D structural information acquired from X-ray CT. In these experiments, frozen samples are imaged in an X-ray CT setup using Medipix based detectors equipped with a CO2 cooled sample holder. Single projections are pre-processed before tomographic reconstruction using a signal-to-thickness calibration. In the second step, the object is sliced into thin sections (circa 20 μm) that are then imaged using both matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and secondary ion (SIMS) mass spectrometry, where the spatial distribution of specific molecules within the sample is determined. The combination of two vastly different imaging approaches provides complementary information (i.e., anatomical and molecular distributions) that allows the correlation of distinct structural features with specific molecules distributions leading to unique insights in disease development.

Position-specific 13C distributions within propane from experiments and natural gas samples

NASA Astrophysics Data System (ADS)

Piasecki, Alison; Sessions, Alex; Lawson, Michael; Ferreira, A. A.; Santos Neto, E. V.; Ellis, Geoffrey S.; Lewan, Michael D.; Eiler, John M.

2018-01-01

Site-specific carbon isotope measurements of organic compounds potentially recover information that is lost in a conventional, 'bulk' isotopic analysis. Such measurements are useful because isotopically fractionating processes may have distinct effects at different molecular sites, and thermodynamically equilibrated populations of molecules tend to concentrate heavy isotopes in one molecular site versus another. Most recent studies of site-specific 13C in organics use specialized Nuclear Magnetic Resonance (NMR) techniques or complex chemical degradations prior to mass spectrometric measurements. Herein we present the first application of a new mass spectrometric technique that reconstructs the site-specific carbon isotope composition of propane based on measurements of the 13C/12C ratios of two or more fragment ions that sample different proportions of the terminal and central carbon sites. We apply this method to propane from laboratory experiments and natural gas samples to explore the relationships between site-specific carbon isotope composition, full-molecular δ13C, thermal maturity, and variation in organic matter precursors. Our goal is to advance the understanding of the sources and histories of short-chain alkanes within geologic systems. Our findings suggest that propane varies in its site-specific carbon isotope structure, which is correlated with increasing thermal maturity, first increasing in terminal position δ13C and then increasing in both center and terminal position δ13C. This pattern is observed in both experimental and natural samples, and is plausibly explained by a combination of site-specific, temperature-dependent isotope effects associated with conversion of different precursor molecules (kerogen, bitumen, and/or oil) to propane, differences in site-specific isotopic contents of those precursors, and possibly distillation of reactive components of those precursors with increasing maturity. We hypothesize that the largest changes in site-specific isotopic content of propane occur when bitumen and/or oil replace kerogen as the dominant precursors. If correct, this phenomenon could have significant utility for understanding gas generation in thermogenic petroleum systems.

Position-specific 13C distributions within propane from experiments and natural gas samples

USGS Publications Warehouse

Piasecki, Alison; Sessions, Alex L.; Lawson, Michael; Ferreira, A.A.; Santos Neto, E. V.; Ellis, Geoffrey S.; Lewan, Michael; Eilers, J.M.

2018-01-01

Site-specific carbon isotope measurements of organic compounds potentially recover information that is lost in a conventional, ‘bulk’ isotopic analysis. Such measurements are useful because isotopically fractionating processes may have distinct effects at different molecular sites, and thermodynamically equilibrated populations of molecules tend to concentrate heavy isotopes in one molecular site versus another. Most recent studies of site-specific 13C in organics use specialized Nuclear Magnetic Resonance (NMR) techniques or complex chemical degradations prior to mass spectrometric measurements. Herein we present the first application of a new mass spectrometric technique that reconstructs the site-specific carbon isotope composition of propane based on measurements of the 13C/12C ratios of two or more fragment ions that sample different proportions of the terminal and central carbon sites. We apply this method to propane from laboratory experiments and natural gas samples to explore the relationships between site-specific carbon isotope composition, full-molecular δ13C, thermal maturity, and variation in organic matter precursors. Our goal is to advance the understanding of the sources and histories of short-chain alkanes within geologic systems. Our findings suggest that propane varies in its site-specific carbon isotope structure, which is correlated with increasing thermal maturity, first increasing in terminal position δ13C and then increasing in both center and terminal position δ13C. This pattern is observed in both experimental and natural samples, and is plausibly explained by a combination of site-specific, temperature-dependent isotope effects associated with conversion of different precursor molecules (kerogen, bitumen, and/or oil) to propane, differences in site-specific isotopic contents of those precursors, and possibly distillation of reactive components of those precursors with increasing maturity. We hypothesize that the largest changes in site-specific isotopic content of propane occur when bitumen and/or oil replace kerogen as the dominant precursors. If correct, this phenomenon could have significant utility for understanding gas generation in thermogenic petroleum systems.

Comparison of techniques for the detection of helminth ova in drinking water and wastewater.

PubMed

Maya, C; Jimenez, B; Schwartzbrod, J

2006-02-01

Many countries use wastewater for irrigation. The World Health Organization established, as reuse guidelines, a maximum value of 1 helminth ovum/L for irrigation. Various techniques for enumerating helminth ova in water have been published. To determine the most adequate method for Mexico, four techniques were compared: the U.S. Environmental Protection Agency (U.S. EPA), membrane-filter, Leeds I, and Faust. Two types of water were used: drinking water and municipal wastewater effluent. Sensitivity, discrimination coefficients, precision, recovery efficiency, and cost were determined. In addition, several unseeded wastewater samples were analyzed. For drinking water, U.S. EPA and the membrane-filter techniques demonstrated comparable results; however, when wastewater was used, the membrane technique showed some deficiencies. Because the U.S. EPA technique may be used for samples with both high and low solids content, allows for the recovery of helminth ova with different specific gravities, and has the lowest total cost, it was selected as the best technique.

Connection between Topic-Specific Teacher Knowledge and Student Performance in Lower Secondary School Mathematics

ERIC Educational Resources Information Center

Tchoshanov, Mourat; Quinones, Maria Cruz; Shakirova, Kadriya B.; Ibragimova, Elena N.; Shakirova, Liliana R.

2017-01-01

The interpretive cross-case study focused on the examination of connections between teacher and student topic-specific knowledge of lower secondary mathematics. Two teachers were selected for the study using non-probability purposive sampling technique. Teachers completed the Teacher Content Knowledge Survey before teaching a topic on division of…

EVALUATE THE UTILITY OF ENTEROCOCCI AS INDICATORS OF THE SOURCES OF FECAL CONTAMINATION IN IMPAIRED SUBWATERSHEDS THROUGH DNA-BASED MOLECULAR TECHNIQUES

EPA Science Inventory

Microbial source tracking (MST) is based on the assumption that specific strains of bacteria are associated with specific host species. MST methods are attractive because their application on environmental samples could help define the nature of water quality problems in impaire...

Application of an oligonucleotide microarray-based nano-amplification technique for the detection of fungal pathogens.

PubMed

Lu, Weiping; Gu, Dayong; Chen, Xingyun; Xiong, Renping; Liu, Ping; Yang, Nan; Zhou, Yuanguo

2010-10-01

The traditional techniques for diagnosis of invasive fungal infections in the clinical microbiology laboratory need improvement. These techniques are prone to delay results due to their time-consuming process, or result in misidentification of the fungus due to low sensitivity or low specificity. The aim of this study was to develop a method for the rapid detection and identification of fungal pathogens. The internal transcribed spacer two fragments of fungal ribosomal DNA were amplified using a polymerase chain reaction for all samples. Next, the products were hybridized with the probes immobilized on the surface of a microarray. These species-specific probes were designed to detect nine different clinical pathogenic fungi including Candida albicans, Candida tropocalis, Candida glabrata, Candida parapsilosis, Candida krusei, Candida lusitaniae, Candida guilliermondii, Candida keyfr, and Cryptococcus neoformans. The hybridizing signals were enhanced with gold nanoparticles and silver deposition, and detected using a flatbed scanner or visually. Fifty-nine strains of fungal pathogens, including standard and clinically isolated strains, were correctly identified by this method. The sensitivity of the assay for Candida albicans was 10 cells/mL. Ten cultures from clinical specimens and 12 clinical samples spiked with fungi were also identified correctly. This technique offers a reliable alternative to conventional methods for the detection and identification of fungal pathogens. It has higher efficiency, specificity and sensitivity compared with other methods commonly used in the clinical laboratory.

Analysis of formalin-fixed, paraffin-embedded (FFPE) tissue via proteomic techniques and misconceptions of antigen retrieval.

PubMed

O'Rourke, Matthew B; Padula, Matthew P

2016-01-01

Since emerging in the late 19(th) century, formaldehyde fixation has become a standard method for preservation of tissues from clinical samples. The advantage of formaldehyde fixation is that fixed tissues can be stored at room temperature for decades without concern for degradation. This has led to the generation of huge tissue banks containing thousands of clinically significant samples. Here we review techniques for proteomic analysis of formalin-fixed, paraffin-embedded (FFPE) tissue samples with a specific focus on the methods used to extract and break formaldehyde crosslinks. We also discuss an error-of-interpretation associated with the technique known as "antigen retrieval." We have discovered that this term has been mistakenly applied to two disparate molecular techniques; therefore, we argue that a terminology change is needed to ensure accurate reporting of experimental results. Finally, we suggest that more investigation is required to fully understand the process of formaldehyde fixation and its subsequent reversal.

Image correlation and sampling study

NASA Technical Reports Server (NTRS)

Popp, D. J.; Mccormack, D. S.; Sedwick, J. L.

1972-01-01

The development of analytical approaches for solving image correlation and image sampling of multispectral data is discussed. Relevant multispectral image statistics which are applicable to image correlation and sampling are identified. The general image statistics include intensity mean, variance, amplitude histogram, power spectral density function, and autocorrelation function. The translation problem associated with digital image registration and the analytical means for comparing commonly used correlation techniques are considered. General expressions for determining the reconstruction error for specific image sampling strategies are developed.

Ambient Mass Spectrometry Imaging Using Direct Liquid Extraction Techniques

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

Laskin, Julia; Lanekoff, Ingela

2015-11-13

Mass spectrometry imaging (MSI) is a powerful analytical technique that enables label-free spatial localization and identification of molecules in complex samples.1-4 MSI applications range from forensics5 to clinical research6 and from understanding microbial communication7-8 to imaging biomolecules in tissues.1, 9-10 Recently, MSI protocols have been reviewed.11 Ambient ionization techniques enable direct analysis of complex samples under atmospheric pressure without special sample pretreatment.3, 12-16 In fact, in ambient ionization mass spectrometry, sample processing (e.g., extraction, dilution, preconcentration, or desorption) occurs during the analysis.17 This substantially speeds up analysis and eliminates any possible effects of sample preparation on the localization of moleculesmore » in the sample.3, 8, 12-14, 18-20 Venter and co-workers have classified ambient ionization techniques into three major categories based on the sample processing steps involved: 1) liquid extraction techniques, in which analyte molecules are removed from the sample and extracted into a solvent prior to ionization; 2) desorption techniques capable of generating free ions directly from substrates; and 3) desorption techniques that produce larger particles subsequently captured by an electrospray plume and ionized.17 This review focuses on localized analysis and ambient imaging of complex samples using a subset of ambient ionization methods broadly defined as “liquid extraction techniques” based on the classification introduced by Venter and co-workers.17 Specifically, we include techniques where analyte molecules are desorbed from solid or liquid samples using charged droplet bombardment, liquid extraction, physisorption, chemisorption, mechanical force, laser ablation, or laser capture microdissection. Analyte extraction is followed by soft ionization that generates ions corresponding to intact species. Some of the key advantages of liquid extraction techniques include the ease of operation, ability to analyze samples in their native environments, speed of analysis, and ability to tune the extraction solvent composition to a problem at hand. For example, solvent composition may be optimized for efficient extraction of different classes of analytes from the sample or for quantification or online derivatization through reactive analysis. In this review, we will: 1) introduce individual liquid extraction techniques capable of localized analysis and imaging, 2) describe approaches for quantitative MSI experiments free of matrix effects, 3) discuss advantages of reactive analysis for MSI experiments, and 4) highlight selected applications (published between 2012 and 2015) that focus on imaging and spatial profiling of molecules in complex biological and environmental samples.« less

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

PubMed

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

2000-01-01

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

[Identification of Vibrio cholerae O1 by flow cytometry].

PubMed

Alvarado-Alemán, F J; González-Bonilla, C; Wong-Arambula, C; Gutiérrez-Cogco, L; Sepúlveda-Amor, J; Kumate-Rodríguez, J

1994-01-01

A total of 72 peptonated water samples suspected of carrying Vibrio cholerae were assessed by laser flow cytometry (LFC) and compared with positive culture. We used a direct fluorescence technique using polyclonal (PolAb) and monoclonal antibodies (MoAb) conjugated to fluorescein. The PolAb were able to detect 33 positive samples. A clear difference among the 20 positive samples was found with only three V. cholerae O1 false negatives when MoAb were used whereas all 13 V. cholerae Non O1 samples were detected. The correlation index comparing control autofluorescence with peptonated water samples show a R = 0.69, versus 0.96 with pure V. cholerae O1 strains. Our data suggest that the LFC technique is able to recognize V. cholerae O1 from a mixture of microorganisms with high sensitivity and specificity in a few hours.

In vivo testing for gold nanoparticle toxicity.

PubMed

Simpson, Carrie A; Huffman, Brian J; Cliffel, David E

2013-01-01

A technique for measuring the toxicity of nanomaterials using a murine model is described. Blood samples are collected via submandibular bleeding while urine samples are collected on cellophane sheets. Both biosamples are then analyzed by inductively coupled plasma optical emission spectroscopy (ICP-OES) for nanotoxicity. Blood samples are further tested for immunological response using a standard Coulter counter. The major organs of interest for filtration are also digested and analyzed via ICP-OES, producing useful information regarding target specificity of the nanomaterial of interest. Collection of the biosamples and analysis afterward is detailed, and the operation of the technique is described and illustrated by analysis of the nanotoxicity of an injection of a modified tiopronin monolayer-protected cluster.

Influence of refrigeration and formalin on the floatability of Giardia duodenalis cysts.

PubMed

Moitinho, M d; Bertoli, M; Guedes, T A; Ferreira, C S

1999-01-01

Giardia duodenalis cysts obtained from fresh fecal samples, fecal samples kept under refrigeration and fecal samples treated with formalin were studied as to their floatability on sucrose solutions with the following specific gravities: 1,040 kg/m3; 1,050 kg/m3; 1, 060 kg/m3; 1,070 kg/m3; 1,080 kg/m3; 1,090 kg/m3; 1,100 kgm3; 1,150 kg/m3; 1,200 kg/m3; and 1,250 kg/m3, contained within counting-chambers 0.17 mm high. Cysts that floated on and those settled down as sediments were counted, and had their percentages estimated. Sucrose solutions of 1,200 kg/m3 specific gravity (the average specific gravity of diluting liquids employed in floatation techniques) caused to float 77.7%, 78.4% and 6.6% of the G. duodenalis cysts obtained, respectively, from fresh fecal samples, fecal samples kept under refrigeration, and fecal samples treated with formalin. Cysts obtained both from fresh fecal samples and fecal samples kept under refrigeration presented similar results concerning floatability. It was observed, however, that the treatment of feces with formalin diminished the cysts floatability under the various specific gravities studied. This results should influence, the recommendations for transport and storage of fecal samples used for parasitological coproscopy.

Classification of bladder cancer cell lines using Raman spectroscopy: a comparison of excitation wavelength, sample substrate and statistical algorithms

NASA Astrophysics Data System (ADS)

Kerr, Laura T.; Adams, Aine; O'Dea, Shirley; Domijan, Katarina; Cullen, Ivor; Hennelly, Bryan M.

2014-05-01

Raman microspectroscopy can be applied to the urinary bladder for highly accurate classification and diagnosis of bladder cancer. This technique can be applied in vitro to bladder epithelial cells obtained from urine cytology or in vivo as an optical biopsy" to provide results in real-time with higher sensitivity and specificity than current clinical methods. However, there exists a high degree of variability across experimental parameters which need to be standardised before this technique can be utilized in an everyday clinical environment. In this study, we investigate different laser wavelengths (473 nm and 532 nm), sample substrates (glass, fused silica and calcium fluoride) and multivariate statistical methods in order to gain insight into how these various experimental parameters impact on the sensitivity and specificity of Raman cytology.

Preparing Monodisperse Macromolecular Samples for Successful Biological Small-Angle X-ray and Neutron Scattering Experiments

PubMed Central

Jeffries, Cy M.; Graewert, Melissa A.; Blanchet, Clément E.; Langley, David B.; Whitten, Andrew E.; Svergun, Dmitri I

2017-01-01

Small-angle X-ray and neutron scattering (SAXS and SANS) are techniques used to extract structural parameters and determine the overall structures and shapes of biological macromolecules, complexes and assemblies in solution. The scattering intensities measured from a sample contain contributions from all atoms within the illuminated sample volume including the solvent and buffer components as well as the macromolecules of interest. In order to obtain structural information, it is essential to prepare an exactly matched solvent blank so that background scattering contributions can be accurately subtracted from the sample scattering to obtain the net scattering from the macromolecules in the sample. In addition, sample heterogeneity caused by contaminants, aggregates, mismatched solvents, radiation damage or other factors can severely influence and complicate data analysis so it is essential that the samples are pure and monodisperse for the duration of the experiment. This Protocol outlines the basic physics of SAXS and SANS and reveals how the underlying conceptual principles of the techniques ultimately ‘translate’ into practical laboratory guidance for the production of samples of sufficiently high quality for scattering experiments. The procedure describes how to prepare and characterize protein and nucleic acid samples for both SAXS and SANS using gel electrophoresis, size exclusion chromatography and light scattering. Also included are procedures specific to X-rays (in-line size exclusion chromatography SAXS) and neutrons, specifically preparing samples for contrast matching/variation experiments and deuterium labeling of proteins. PMID:27711050

Use of ProteinChip technology for identifying biomarkers of parasitic diseases: the example of porcine cysticercosis (Taenia solium).

PubMed

Deckers, N; Dorny, P; Kanobana, K; Vercruysse, J; Gonzalez, A E; Ward, B; Ndao, M

2008-12-01

Taenia solium cysticercosis is a significant public health problem in endemic countries. The current serodiagnostic techniques are not able to differentiate between infections with viable cysts and infections with degenerated cysts. The objectives of this study were to identify specific novel biomarkers of these different disease stages in the serum of experimentally infected pigs using ProteinChip technology (Bio-Rad) and to validate these biomarkers by analyzing serum samples from naturally infected pigs. In the experimental sample set 30 discriminating biomarkers (p<0.05) were found, 13 specific for the viable phenotype, 9 specific for the degenerated phenotype and 8 specific for the infected phenotype (either viable or degenerated cysts). Only 3 of these biomarkers were also significant in the field samples; however, the peak profiles were not consistent among the two sample sets. Five biomarkers discovered in the sera from experimentally infected pigs were identified as clusterin, lecithin-cholesterol acyltransferase, vitronectin, haptoglobin and apolipoprotein A-I.

StreamCat and LakeCat: An overview of algorithms, data, and models developed at the US EPA Western Ecology Division to facilitate and advance watershed prediction in the conterminous US.

EPA Science Inventory

Geospatial data and techniques have long been critical to advancing the analysis and management of freshwater ecosystems. However, these data and techniques have often been limited to specific sample sites or regional analyses because of the difficulty associated with generating ...

Accelerated spike resampling for accurate multiple testing controls.

PubMed

Harrison, Matthew T

2013-02-01

Controlling for multiple hypothesis tests using standard spike resampling techniques often requires prohibitive amounts of computation. Importance sampling techniques can be used to accelerate the computation. The general theory is presented, along with specific examples for testing differences across conditions using permutation tests and for testing pairwise synchrony and precise lagged-correlation between many simultaneously recorded spike trains using interval jitter.

Guidelines for collection and field analysis of ground-water samples for selected unstable constituents

USGS Publications Warehouse

Wood, Warren W.

1976-01-01

The unstable nature of many chemical and physical constituents in ground water requires special collection procedures and field analysis immediately after collection. This report describes the techniques and equipment commonly used m the collection and field analysis of samples for pH, temperature, carbonate, bicarbonate, specific conductance, Eh, and dissolved oxygen.

Using Sieving and Unknown Sand Samples for a Sedimentation-Stratigraphy Class Project with Linkage to Introductory Courses

ERIC Educational Resources Information Center

Videtich, Patricia E.; Neal, William J.

2012-01-01

Using sieving and sample "unknowns" for instructional grain-size analysis and interpretation of sands in undergraduate sedimentology courses has advantages over other techniques. Students (1) learn to calculate and use statistics; (2) visually observe differences in the grain-size fractions, thereby developing a sense of specific size…

A novel approach to enhance antibody sensitivity and specificity by peptide cross-linking

PubMed Central

Namiki, Takeshi; Valencia, Julio C.; Hall, Matthew D.; Hearing, Vincent J.

2008-01-01

Most current techniques employed to improve antigen-antibody signals in western blotting and in immunohistochemistry rely on sample processing prior to staining (e.g. microwaving) or using a more robust reporter (e.g. a secondary antibody with biotin-streptavidin). We have developed and optimized a new approach intended to stabilize the complexes formed between antigens and their respective primary antibodies by cupric ions at high pH. This technique improves the affinity and lowers cross-reactivity with non-specific bands of ∼20% of antibodies tested (5/25). Here we report that this method can enhance antigen-antibody specificity and can improve the utility of some poorly reactive primary antibodies. PMID:18801330

The use of multilevel sampling techniques for determining shallow aquifer nitrate profiles.

PubMed

Lasagna, Manuela; De Luca, Domenico Antonio

2016-10-01

Nitrate is a worldwide pollutant in aquifers. Shallow aquifer nitrate concentrations generally display vertical stratification, with a maximum concentration immediately below the water level. The concentration then gradually decreases with depth. Different techniques can be used to highlight this stratification. The paper aims at comparing the advantages and limitations of three open hole multilevel sampling techniques (packer system, dialysis membrane samplers and bailer), chosen on the base of a literary review, to highlight a nitrate vertical stratification under the assumption of (sub)horizontal flow in the aquifer. The sampling systems were employed at three different times of the year in a shallow aquifer piezometer in northern Italy. The optimal purge time, equilibration time and water volume losses during the time in the piezometer were evaluated. Multilevel techniques highlighted a similar vertical nitrate stratification, present throughout the year. Indeed, nitrate concentrations generally decreased with depth downwards, but with significantly different levels in the sampling campaigns. Moreover, the sampling techniques produced different degrees of accuracy. More specifically, the dialysis membrane samplers provided the most accurate hydrochemical profile of the shallow aquifer and they appear to be necessary when the objective is to detect the discontinuities in the nitrate profile. Bailer and packer system showed the same nitrate profile with little differences of concentration. However, the bailer resulted much more easier to use.

High-resolution X-ray diffraction with no sample preparation

PubMed Central

Turner, S. M. R.; Degryse, P.; Shortland, A. J.

2017-01-01

It is shown that energy-dispersive X-ray diffraction (EDXRD) implemented in a back-reflection geometry is extremely insensitive to sample morphology and positioning even in a high-resolution configuration. This technique allows high-quality X-ray diffraction analysis of samples that have not been prepared and is therefore completely non-destructive. The experimental technique was implemented on beamline B18 at the Diamond Light Source synchrotron in Oxfordshire, UK. The majority of the experiments in this study were performed with pre-characterized geological materials in order to elucidate the characteristics of this novel technique and to develop the analysis methods. Results are presented that demonstrate phase identification, the derivation of precise unit-cell parameters and extraction of microstructural information on unprepared rock samples and other sample types. A particular highlight was the identification of a specific polytype of a muscovite in an unprepared mica schist sample, avoiding the time-consuming and difficult preparation steps normally required to make this type of identification. The technique was also demonstrated in application to a small number of fossil and archaeological samples. Back-reflection EDXRD implemented in a high-resolution configuration shows great potential in the crystallographic analysis of cultural heritage artefacts for the purposes of scientific research such as provenancing, as well as contributing to the formulation of conservation strategies. Possibilities for moving the technique from the synchrotron into museums are discussed. The avoidance of the need to extract samples from high-value and rare objects is a highly significant advantage, applicable also in other potential research areas such as palaeontology, and the study of meteorites and planetary materials brought to Earth by sample-return missions. PMID:28660862

Reaching multi-nanosecond timescales in combined QM/MM molecular dynamics simulations through parallel horsetail sampling.

PubMed

Martins-Costa, Marilia T C; Ruiz-López, Manuel F

2017-04-15

We report an enhanced sampling technique that allows to reach the multi-nanosecond timescale in quantum mechanics/molecular mechanics molecular dynamics simulations. The proposed technique, called horsetail sampling, is a specific type of multiple molecular dynamics approach exhibiting high parallel efficiency. It couples a main simulation with a large number of shorter trajectories launched on independent processors at periodic time intervals. The technique is applied to study hydrogen peroxide at the water liquid-vapor interface, a system of considerable atmospheric relevance. A total simulation time of a little more than 6 ns has been attained for a total CPU time of 5.1 years representing only about 20 days of wall-clock time. The discussion of the results highlights the strong influence of the solvation effects at the interface on the structure and the electronic properties of the solute. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Rapid analysis of adulterations in Chinese lotus root powder (LRP) by near-infrared (NIR) spectroscopy coupled with chemometric class modeling techniques.

PubMed

Xu, Lu; Shi, Peng-Tao; Ye, Zi-Hong; Yan, Si-Min; Yu, Xiao-Ping

2013-12-01

This paper develops a rapid analysis method for adulteration identification of a popular traditional Chinese food, lotus root powder (LRP), by near-infrared spectroscopy and chemometrics. 85 pure LRP samples were collected from 7 main lotus producing areas of China to include most if not all of the significant variations likely to be encountered in unknown authentic materials. To evaluate the model specificity, 80 adulterated LRP samples prepared by blending pure LRP with different levels of four cheaper and commonly used starches were measured and predicted. For multivariate quality models, two class modeling methods, the traditional soft independent modeling of class analogy (SIMCA) and a recently proposed partial least squares class model (PLSCM) were used. Different data preprocessing techniques, including smoothing, taking derivative and standard normal variate (SNV) transformation were used to improve the classification performance. The results indicate that smoothing, taking second-order derivatives and SNV can improve the class models by enhancing signal-to-noise ratio, reducing baseline and background shifts. The most accurate and stable models were obtained with SNV spectra for both SIMCA (sensitivity 0.909 and specificity 0.938) and PLSCM (sensitivity 0.909 and specificity 0.925). Moreover, both SIMCA and PLSCM could detect LRP samples mixed with 5% (w/w) or more other cheaper starches, including cassava, sweet potato, potato and maize starches. Although it is difficult to perform an exhaustive collection of all pure LRP samples and possible adulterations, NIR spectrometry combined with class modeling techniques provides a reliable and effective method to detect most of the current LRP adulterations in Chinese market. Copyright © 2013 Elsevier Ltd. All rights reserved.

Specific Features of Social Competence Development in the Future Music Teachers Working at Universities

ERIC Educational Resources Information Center

Dzheksembekova, Menslu I.; Ibrayeva, Kamarsulu E.; Akhmetova, Aimkul K.; Urazalieva, Moldir A.; Sultangaliyeva, Elmira S.; Issametova, Klavdiya I.

2016-01-01

This paper aims at analyzing specific features of social competence of future music teachers and the development of specialized techniques in order to improve the quality of motivational and cognitive components of student social competence. The sample involved 660 undergraduate students. The authors used a number of research methods, such as…

EVALUATE THE UTILITY OF ENTEROCOCCI AND BACTEROIDES AS INDICATORS OF THE SOURCES OF FECAL CONTAMINATION IN IMPAIRED SUBWATERSHEDS THROUGH DNA-BASED MOLECULAR TECHNIQUES.

EPA Science Inventory

Microbial source tracking (MST) is based on the assumption that specific strains of bacteria are associated with specific host species. MST methods are attractive because their application on environmental samples could help define the nature of water quality problems in impaire...

Recent progress of task-specific ionic liquids in chiral resolution and extraction of biological samples and metal ions.

PubMed

Wu, Datong; Cai, Pengfei; Zhao, Xiaoyong; Kong, Yong; Pan, Yuanjiang

2018-01-01

Ionic liquids have been functionalized for modern applications. The functional ionic liquids are also called task-specific ionic liquids. Various task-specific ionic liquids with certain groups have been constructed and exploited widely in the field of separation. To take advantage of their properties in separation science, task-specific ionic liquids are generally used in techniques such as liquid-liquid extraction, solid-phase extraction, gas chromatography, high-performance liquid chromatography, and capillary electrophoresis. This review mainly covers original research papers published in the last five years, and we will focus on task-specific ionic liquids as the chiral selectors in chiral resolution and as extractant or sensor for biological samples and metal ion purification. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Assessing the spatial distribution of glyphosate-AMPA in an Argentinian farm field using a pedometric technique

NASA Astrophysics Data System (ADS)

Barbera, Agustin; Zamora, Martin; Domenech, Marisa; Vega-Becerra, Andres; Castro-Franco, Mauricio

2017-04-01

The cultivation of transgenic glyphosate-resistant crops has been the most rapidly adopted crop technology in Argentina since 1997. Thus, more than 180 million liters of the broad-spectrum herbicide glyphosate (N - phosphonomethylglicine) are applied every year. The intensive use of glyphosate combined with geomorphometrical characteristics of the Pampa region is a matter of environmental concern. An integral component of assessing the risk of soil contamination in farm fields is to describe the spatial distribution of the levels of contaminant agent. Application of pedometric techniques for this purpose has been scarcely demonstrated. These techniques could provide an estimate of the concentration at a given unsampled location, as well as the probability that concentration will exceed the critical threshold concentration. In this work, a pedometric technique for assessing the spatial distribution of glyphosate in farm fields was developed. A field located at INTA Barrow, Argentina (Lat: -38.322844, Lon: -60.25572) which has a great soil spatial variability, was divided by soil-specific zones using a pedometric technique. This was developed integrating INTA Soil Survey information and a digital elevation model (DEM) obtained from a DGPS. Firstly, 10 topographic indices derived from a DEM were computed in a Random Forest algorithm to obtain a classification model for soil map units (SMU). Secondly, a classification model was applied to those topographic indices but at a scale higher than 1:1000. Finally, a spatial principal component analysis and a clustering using Fuzzy K-means were used into each SMU. From this clustering, three soil-specific zones were determined which were also validated through apparent electrical conductivity (CEa) measurements. Three soil sample points were determined by zone. In each one, samples from 0-10, 10-20 and 20-40cm depth were taken. Glyphosate content and AMPA in each soil sample were analyzed using de UPLC-MS/MS ESI (+/-). Only AMPA at 10-20 cm depth had significant difference among soil-specific zones. However, marked trends for glyphosate content and AMPA were clearly shown among zones. These results suggest that (i) the presence of glyphosate and AMPA has spatial patterns distribution related to soil properties at field scale; and (ii) the proposed technique allowed to determine soil-specific zones related to the spatial distribution of glyphosate and AMPA fast, cost-effective and accurately. In further works, we would suggest adding new soil information sources to improve soil-specific zone delimitation.

Concentration and separation of biological organisms by ultrafiltration and dielectrophoresis

DOEpatents

Simmons, Blake A.; Hill, Vincent R.; Fintschenko, Yolanda; Cummings, Eric B.

2010-10-12

Disclosed is a method for monitoring sources of public water supply for a variety of pathogens by using a combination of ultrafiltration techniques together dielectrophoretic separation techniques. Because water-borne pathogens, whether present due to "natural" contamination or intentional introduction, would likely be present in drinking water at low concentrations when samples are collected for monitoring or outbreak investigations, an approach is needed to quickly and efficiently concentrate and separate particles such as viruses, bacteria, and parasites in large volumes of water (e.g., 100 L or more) while simultaneously reducing the sample volume to levels sufficient for detecting low concentrations of microbes (e.g., <10 mL). The technique is also designed to screen the separated microbes based on specific conductivity and size.

Method And Apparatus For Evaluatin Of High Temperature Superconductors

DOEpatents

Fishman, Ilya M.; Kino, Gordon S.

1996-11-12

A technique for evaluation of high-T.sub.c superconducting films and single crystals is based on measurement of temperature dependence of differential optical reflectivity of high-T.sub.c materials. In the claimed method, specific parameters of the superconducting transition such as the critical temperature, anisotropy of the differential optical reflectivity response, and the part of the optical losses related to sample quality are measured. The apparatus for performing this technique includes pump and probe sources, cooling means for sweeping sample temperature across the critical temperature and polarization controller for controlling a state of polarization of a probe light beam.

The signature-based radiation-scanning approach to standoff detection of improvised explosive devices.

PubMed

Brewer, R L; Dunn, W L; Heider, S; Matthew, C; Yang, X

2012-07-01

The signature-based radiation-scanning technique for detection of improvised explosive devices is described. The technique seeks to detect nitrogen-rich chemical explosives present in a target. The technology compares a set of "signatures" obtained from a test target to a collection of "templates", sets of signatures for a target that contain an explosive in a specific configuration. Interrogation of nitrogen-rich fertilizer samples, which serve as surrogates for explosives, is shown experimentally to be able to discriminate samples of 3.8L and larger. Copyright © 2011 Elsevier Ltd. All rights reserved.

Programmed LWR metrology by multi-techniques approach

NASA Astrophysics Data System (ADS)

Reche, Jérôme; Besacier, Maxime; Gergaud, Patrice; Blancquaert, Yoann; Freychet, Guillaume; Labbaye, Thibault

2018-03-01

Nowadays, roughness control presents a huge challenge for the lithography step. For advanced nodes, this morphological aspect reaches the same order of magnitude than the Critical Dimension. Hence, the control of roughness needs an adapted metrology. In this study, specific samples with designed roughness have been manufactured using e-beam lithography. These samples have been characterized with three different methodologies: CD-SEM, OCD and SAXS. The main goal of the project is to compare the capability of each of these techniques in terms of reliability, type of information obtained, time to obtain the measurements and level of maturity for the industry.

Sampling Methods in Cardiovascular Nursing Research: An Overview.

PubMed

Kandola, Damanpreet; Banner, Davina; O'Keefe-McCarthy, Sheila; Jassal, Debbie

2014-01-01

Cardiovascular nursing research covers a wide array of topics from health services to psychosocial patient experiences. The selection of specific participant samples is an important part of the research design and process. The sampling strategy employed is of utmost importance to ensure that a representative sample of participants is chosen. There are two main categories of sampling methods: probability and non-probability. Probability sampling is the random selection of elements from the population, where each element of the population has an equal and independent chance of being included in the sample. There are five main types of probability sampling including simple random sampling, systematic sampling, stratified sampling, cluster sampling, and multi-stage sampling. Non-probability sampling methods are those in which elements are chosen through non-random methods for inclusion into the research study and include convenience sampling, purposive sampling, and snowball sampling. Each approach offers distinct advantages and disadvantages and must be considered critically. In this research column, we provide an introduction to these key sampling techniques and draw on examples from the cardiovascular research. Understanding the differences in sampling techniques may aid nurses in effective appraisal of research literature and provide a reference pointfor nurses who engage in cardiovascular research.

Melting temperature and enthalpy variations of phase change materials (PCMs): a differential scanning calorimetry (DSC) analysis

NASA Astrophysics Data System (ADS)

Sun, Xiaoqin; Lee, Kyoung Ok; Medina, Mario A.; Chu, Youhong; Li, Chuanchang

2018-06-01

Differential scanning calorimetry (DSC) analysis is a standard thermal analysis technique used to determine the phase transition temperature, enthalpy, heat of fusion, specific heat and activation energy of phase change materials (PCMs). To determine the appropriate heating rate and sample mass, various DSC measurements were carried out using two kinds of PCMs, namely N-octadecane paraffin and calcium chloride hexahydrate. The variations in phase transition temperature, enthalpy, heat of fusion, specific heat and activation energy were observed within applicable heating rates and sample masses. It was found that the phase transition temperature range increased with increasing heating rate and sample mass; while the heat of fusion varied without any established pattern. The specific heat decreased with the increase of heating rate and sample mass. For accuracy purpose, it is recommended that for PCMs with high thermal conductivity (e.g. hydrated salt) the focus will be on heating rate rather than sample mass.

A grid matrix-based Raman spectroscopic method to characterize different cell milieu in biopsied axillary sentinel lymph nodes of breast cancer patients.

PubMed

Som, Dipasree; Tak, Megha; Setia, Mohit; Patil, Asawari; Sengupta, Amit; Chilakapati, C Murali Krishna; Srivastava, Anurag; Parmar, Vani; Nair, Nita; Sarin, Rajiv; Badwe, R

2016-01-01

Raman spectroscopy which is based upon inelastic scattering of photons has a potential to emerge as a noninvasive bedside in vivo or ex vivo molecular diagnostic tool. There is a need to improve the sensitivity and predictability of Raman spectroscopy. We developed a grid matrix-based tissue mapping protocol to acquire cellular-specific spectra that also involved digital microscopy for localizing malignant and lymphocytic cells in sentinel lymph node biopsy sample. Biosignals acquired from specific cellular milieu were subjected to an advanced supervised analytical method, i.e., cross-correlation and peak-to-peak ratio in addition to PCA and PC-LDA. We observed decreased spectral intensity as well as shift in the spectral peaks of amides and lipid bands in the completely metastatic (cancer cells) lymph nodes with high cellular density. Spectral library of normal lymphocytes and metastatic cancer cells created using the cellular specific mapping technique can be utilized to create an automated smart diagnostic tool for bench side screening of sampled lymph nodes. Spectral library of normal lymphocytes and metastatic cancer cells created using the cellular specific mapping technique can be utilized to develop an automated smart diagnostic tool for bench side screening of sampled lymph nodes supported by ongoing global research in developing better technology and signal and big data processing algorithms.

Approaches to Recruiting ‘Hard-To-Reach’ Populations into Re­search: A Review of the Literature

PubMed Central

Shaghaghi, Abdolreza; Bhopal, Raj S; Sheikh, Aziz

2011-01-01

Background: ‘Hard-to-reach’ is a term used to describe those sub-groups of the population that may be difficult to reach or involve in research or public health programmes. Application of a single term to call these sub-sections of populations implies a homogeneity within distinct groups, which does not necessarily exist. Different sampling techniques were introduced so far to recruit hard-to-reach populations. In this article, we have reviewed a range of ap­proaches that have been used to widen participation in studies. Methods: We performed a Pubmed and Google search for relevant English language articles using the keywords and phrases: (hard-to-reach AND population* OR sampl*), (hidden AND population* OR sample*) and (“hard to reach” AND population* OR sample*) and a consul­tation of the retrieved articles’ bibliographies to extract empirical evidence from publications that discussed or examined the use of sampling techniques to recruit hidden or hard-to-reach populations in health studies. Results: Reviewing the literature has identified a range of techniques to recruit hard-to-reach populations, including snowball sampling, respondent-driven sampling (RDS), indigenous field worker sampling (IFWS), facility-based sampling (FBS), targeted sampling (TS), time-location (space) sampling (TLS), conventional cluster sampling (CCS) and capture re-capture sampling (CR). Conclusion: The degree of compliance with a study by a certain ‘hard-to-reach’ group de­pends on the characteristics of that group, recruitment technique used and the subject of inter­est. Irrespective of potential advantages or limitations of the recruitment techniques reviewed, their successful use depends mainly upon our knowledge about specific characteristics of the target populations. Thus in line with attempts to expand the current boundaries of our know­ledge about recruitment techniques in health studies and their applications in varying situa­tions, we should also focus on possibly all contributing factors which may have an impact on participation rate within a defined population group. PMID:24688904

Promotional methods used by representatives of drug companies: A prospective survey in general practice

PubMed Central

Schramm, Jesper; Andersen, Morten; Vach, Kirstin; Kragstrup, Jakob; Peter Kampmann, Jens; Søndergaard, Jens

2007-01-01

Objective To examine the extent and composition of pharmaceutical industry representatives’ marketing techniques with a particular focus on drug sampling in relation to drug age. Design A group of 47 GPs prospectively collected data on drug promotional activities during a six-month period, and a sub-sample of 10 GPs furthermore recorded the representatives’ marketing techniques in detail. Setting Primary healthcare. Subjects General practitioners in the County of Funen, Denmark. Main outcome measures. Promotional visits and corresponding marketing techniques. Results The 47 GPs recorded 1050 visits corresponding to a median of 19 (range 3 to 63) per GP in the six months. The majority of drugs promoted (52%) were marketed more than five years ago. There was a statistically significant decline in the proportion of visits where drug samples were offered with drug age, but the decline was small OR 0.97 (95% CI 0.95;0.98) per year. Leaflets (68%), suggestions on how to improve therapy for a specific patient registered with the practice (53%), drug samples (48%), and gifts (36%) were the most frequently used marketing techniques. Conclusion Drug-industry representatives use a variety of promotional methods. The tendency to hand out drug samples was statistically significantly associated with drug age, but the decline was small. PMID:17497486

Development and potential applications of microarrays based on fluorescent nanocrystal-encoded beads for multiplexed cancer diagnostics

NASA Astrophysics Data System (ADS)

Brazhnik, Kristina; Grinevich, Regina; Efimov, Anton E.; Nabiev, Igor; Sukhanova, Alyona

2014-05-01

Advanced multiplexed assays have recently become an indispensable tool for clinical diagnostics. These techniques provide simultaneous quantitative determination of multiple biomolecules in a single sample quickly and accurately. The development of multiplex suspension arrays is currently of particular interest for clinical applications. Optical encoding of microparticles is the most available and easy-to-use technique. This technology uses fluorophores incorporated into microbeads to obtain individual optical codes. Fluorophore-encoded beads can be rapidly analyzed using classical flow cytometry or microfluidic techniques. We have developed a new generation of highly sensitive and specific diagnostic systems for detection of cancer antigens in human serum samples based on microbeads encoded with fluorescent quantum dots (QDs). The designed suspension microarray system was validated for quantitative detection of (1) free and total prostate specific antigen (PSA) in the serum of patients with prostate cancer and (2) carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA 15-3) in the serum of patients with breast cancer. The serum samples from healthy donors were used as a control. The antigen detection is based on the formation of an immune complex of a specific capture antibody (Ab), a target antigen (Ag), and a detector Ab on the surface of the encoded particles. The capture Ab is bound to the polymer shell of microbeads via an adapter molecule, for example, protein A. Protein A binds a monoclonal Ab in a highly oriented manner due to specific interaction with the Fc-region of the Ab molecule. Each antigen can be recognized and detected due to a specific microbead population carrying the unique fluorescent code. 100 and 231 serum samples from patients with different stages of prostate cancer and breast cancer, respectively, and those from healthy donors were examined using the designed suspension system. The data were validated by comparing with the results of the "gold standard" enzyme-linked immunosorbent assay (ELISA). They have shown that our approach is a good alternative to the diagnostics of cancer markers using conventional assays, especially in early diagnostic applications.

Measuring molecular biomarkers in epidemiologic studies: laboratory techniques and biospecimen considerations.

PubMed

Erickson, Heidi S

2012-09-28

The future of personalized medicine depends on the ability to efficiently and rapidly elucidate a reliable set of disease-specific molecular biomarkers. High-throughput molecular biomarker analysis methods have been developed to identify disease risk, diagnostic, prognostic, and therapeutic targets in human clinical samples. Currently, high throughput screening allows us to analyze thousands of markers from one sample or one marker from thousands of samples and will eventually allow us to analyze thousands of markers from thousands of samples. Unfortunately, the inherent nature of current high throughput methodologies, clinical specimens, and cost of analysis is often prohibitive for extensive high throughput biomarker analysis. This review summarizes the current state of high throughput biomarker screening of clinical specimens applicable to genetic epidemiology and longitudinal population-based studies with a focus on considerations related to biospecimens, laboratory techniques, and sample pooling. Copyright © 2012 John Wiley & Sons, Ltd.

Simulating and assessing boson sampling experiments with phase-space representations

NASA Astrophysics Data System (ADS)

Opanchuk, Bogdan; Rosales-Zárate, Laura; Reid, Margaret D.; Drummond, Peter D.

2018-04-01

The search for new, application-specific quantum computers designed to outperform any classical computer is driven by the ending of Moore's law and the quantum advantages potentially obtainable. Photonic networks are promising examples, with experimental demonstrations and potential for obtaining a quantum computer to solve problems believed classically impossible. This introduces a challenge: how does one design or understand such photonic networks? One must be able to calculate observables using general methods capable of treating arbitrary inputs, dissipation, and noise. We develop complex phase-space software for simulating these photonic networks, and apply this to boson sampling experiments. Our techniques give sampling errors orders of magnitude lower than experimental correlation measurements for the same number of samples. We show that these techniques remove systematic errors in previous algorithms for estimating correlations, with large improvements in errors in some cases. In addition, we obtain a scalable channel-combination strategy for assessment of boson sampling devices.

Facile synthesis of Li2S-P2S5 glass-ceramics electrolyte with micron range particles for all-solid-state batteries via a low-temperature solution technique (LTST)

NASA Astrophysics Data System (ADS)

Choi, Sunho; Lee, Sewook; Park, Jongyeop; Nichols, William T.; Shin, Dongwook

2018-06-01

A lithium ion conductive 75Li2Sṡ25P2S5 glass-ceramics electrolyte is, for the first time, successfully synthesized via a new low-temperature solution technique (LTST) and compared to the conventional mechanical-milling technique. Both samples are composed of the highly lithium ion conductive thio-LISICON III analog phase. Due to the uniform dispersion of reactants in an organic liquid, the use of LTST produced significantly smaller and more uniform particle sizes (2.2 ± 1.68 μm) resulting in a 6.5 times higher specific surface area compared to the mechanically-milled sample. A pronounced enhancement of both the rate capability and cyclability is demonstrated for the LTST solid electrolyte sample due to the more intimate contact with the LiCoO2 active material. Furthermore, the LTST sample shows excellent electrochemical stability throughout the potential range of -1 to 5 V. These results suggest that the proposed technique using the optimized LTST process is promising for the preparation of 75Li2Sṡ25P2S5 solid electrolytes for use in advanced Li-ion batteries.

Applications of surface analytical techniques in Earth Sciences

NASA Astrophysics Data System (ADS)

Qian, Gujie; Li, Yubiao; Gerson, Andrea R.

2015-03-01

This review covers a wide range of surface analytical techniques: X-ray photoelectron spectroscopy (XPS), scanning photoelectron microscopy (SPEM), photoemission electron microscopy (PEEM), dynamic and static secondary ion mass spectroscopy (SIMS), electron backscatter diffraction (EBSD), atomic force microscopy (AFM). Others that are relatively less widely used but are also important to the Earth Sciences are also included: Auger electron spectroscopy (AES), low energy electron diffraction (LEED) and scanning tunnelling microscopy (STM). All these techniques probe only the very top sample surface layers (sub-nm to several tens of nm). In addition, we also present several other techniques i.e. Raman microspectroscopy, reflection infrared (IR) microspectroscopy and quantitative evaluation of minerals by scanning electron microscopy (QEMSCAN) that penetrate deeper into the sample, up to several μm, as all of them are fundamental analytical tools for the Earth Sciences. Grazing incidence synchrotron techniques, sensitive to surface measurements, are also briefly introduced at the end of this review. (Scanning) transmission electron microscopy (TEM/STEM) is a special case that can be applied to characterisation of mineralogical and geological sample surfaces. Since TEM/STEM is such an important technique for Earth Scientists, we have also included it to draw attention to the capability of TEM/STEM applied as a surface-equivalent tool. While this review presents most of the important techniques for the Earth Sciences, it is not an all-inclusive bibliography of those analytical techniques. Instead, for each technique that is discussed, we first give a very brief introduction about its principle and background, followed by a short section on approaches to sample preparation that are important for researchers to appreciate prior to the actual sample analysis. We then use examples from publications (and also some of our known unpublished results) within the Earth Sciences to show how each technique is applied and used to obtain specific information and to resolve real problems, which forms the central theme of this review. Although this review focuses on applications of these techniques to study mineralogical and geological samples, we also anticipate that researchers from other research areas such as Material and Environmental Sciences may benefit from this review.

Isotope ratio analysis by Orbitrap mass spectrometry

NASA Astrophysics Data System (ADS)

Eiler, J. M.; Chimiak, L. M.; Dallas, B.; Griep-Raming, J.; Juchelka, D.; Makarov, A.; Schwieters, J. B.

2016-12-01

Several technologies are being developed to examine the intramolecular isotopic structures of molecules (i.e., site-specific and multiple substitution), but various limitations in sample size and type or (for IRMS) resolution have so far prevented the creation of a truly general technique. We will discuss the initial findings of a technique based on Fourier transform mass spectrometry, using the Thermo Scientific Q Exactive GC — an instrument that contains an Orbitrap mass analyzer. Fourier transform mass spectrometry is marked by exceptionally high mass resolutions (the Orbitrap reaches M/ΔM in the range 250,000-1M in the mass range of greatest interest, 50-200 amu). This allows for resolution of a large range of nearly isobaric interferences for isotopologues of volatile and semi-volatile compounds (i.e., involving isotopes of H, C, N, O and S). It also provides potential to solve very challenging mass resolution problems for isotopic analysis of other, heavier elements. Both internal and external experimental reproducibilities of isotope ratio analyses using the Orbitrap typically conform to shot-noise limits down to levels of 0.2 ‰ (1SE), and routinely in the range 0.5-1.0 ‰, with similar accuracy when standardized to concurrently run reference materials. Such measurements can be made without modifications to the ion optics of the Q Exactive GC, but do require specially designed sample introduction devices to permit sample/standard comparison and long integration times. The sensitivity of the Q Exactive GC permits analysis of sub-nanomolar samples and quantification of multiply-substituted species. The site-specific capability of this instrument arises from the fact that mass spectra of molecular analytes commonly contain diverse fragment ion species, each of which samples a specific sub-set of molecular sites. We will present applications of this technique to the biological and abiological chemistry of amino acids, forensic identification of hydrocarbon environmental pollutants, and study of the origins of isotope anomalies in meteoritic organics.

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.

Application of the angle measure technique as image texture analysis method for the identification of uranium ore concentrate samples: New perspective in nuclear forensics.

PubMed

Fongaro, Lorenzo; Ho, Doris Mer Lin; Kvaal, Knut; Mayer, Klaus; Rondinella, Vincenzo V

2016-05-15

The identification of interdicted nuclear or radioactive materials requires the application of dedicated techniques. In this work, a new approach for characterizing powder of uranium ore concentrates (UOCs) is presented. It is based on image texture analysis and multivariate data modelling. 26 different UOCs samples were evaluated applying the Angle Measure Technique (AMT) algorithm to extract textural features on samples images acquired at 250× and 1000× magnification by Scanning Electron Microscope (SEM). At both magnifications, this method proved effective to classify the different types of UOC powder based on the surface characteristics that depend on particle size, homogeneity, and graininess and are related to the composition and processes used in the production facilities. Using the outcome data from the application of the AMT algorithm, the total explained variance was higher than 90% with Principal Component Analysis (PCA), while partial least square discriminant analysis (PLS-DA) applied only on the 14 black colour UOCs powder samples, allowed their classification only on the basis of their surface texture features (sensitivity>0.6; specificity>0.6). This preliminary study shows that this method was able to distinguish samples with similar composition, but obtained from different facilities. The mean angle spectral data obtained by the image texture analysis using the AMT algorithm can be considered as a specific fingerprint or signature of UOCs and could be used for nuclear forensic investigation. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

GC/MS ANALYSIS OF PCB CONGENERS IN BLOOD OF THE HARBOR SEAL PHOCA VITULINA FROM SAN FRANCISCO BAY

EPA Science Inventory

Here we report a validated technique for quantifying up to 20 specific PCB congeners in 1-2 g samples of whole blood with a detection limit below 1 ng/g (ppb) wet weight. Specimens were analyzed from 14 harbor seals sampled in south San Francisco Bay, California during 1991-1992....

Semiautomated confocal imaging of fungal pathogenesis on plants: Microscopic analysis of macroscopic specimens.

PubMed

Minker, Katharine R; Biedrzycki, Meredith L; Kolagunda, Abhishek; Rhein, Stephen; Perina, Fabiano J; Jacobs, Samuel S; Moore, Michael; Jamann, Tiffany M; Yang, Qin; Nelson, Rebecca; Balint-Kurti, Peter; Kambhamettu, Chandra; Wisser, Randall J; Caplan, Jeffrey L

2018-02-01

The study of phenotypic variation in plant pathogenesis provides fundamental information about the nature of disease resistance. Cellular mechanisms that alter pathogenesis can be elucidated with confocal microscopy; however, systematic phenotyping platforms-from sample processing to image analysis-to investigate this do not exist. We have developed a platform for 3D phenotyping of cellular features underlying variation in disease development by fluorescence-specific resolution of host and pathogen interactions across time (4D). A confocal microscopy phenotyping platform compatible with different maize-fungal pathosystems (fungi: Setosphaeria turcica, Cochliobolus heterostrophus, and Cercospora zeae-maydis) was developed. Protocols and techniques were standardized for sample fixation, optical clearing, species-specific combinatorial fluorescence staining, multisample imaging, and image processing for investigation at the macroscale. The sample preparation methods presented here overcome challenges to fluorescence imaging such as specimen thickness and topography as well as physiological characteristics of the samples such as tissue autofluorescence and presence of cuticle. The resulting imaging techniques provide interesting qualitative and quantitative information not possible with conventional light or electron 2D imaging. Microsc. Res. Tech., 81:141-152, 2018. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Microbial composition of the Korean traditional food "kochujang" analyzed by a massive sequencing technique.

PubMed

Nam, Young-Do; Park, So-lim; Lim, Seong-Il

2012-04-01

Kochujang is a traditional Korean fermented food that is made with red pepper, glutinous rice, salt, and soybean. Kochujang is fermented by naturally occurring microorganisms through which it obtains various health-promoting properties. In this study, the bacterial diversities of 9 local and 2 commercial brands of kochujang were analyzed with a barcoded pyrosequencing technique targeting the hyper-variable regions V1/V2 of the 16S rRNA gene. Through the analysis of 13524 bacterial pyrosequences, 223 bacterial species were identified, most of which converged on the phylum Firmicutes (average 93.1%). All of the kochujang samples were largely populated (>90.9% of abundance) by 12 bacterial families, and Bacillaceae showed the highest abundance in all but one sample. Bacillus subtilis and B. licheniformis were the most dominant bacterial species and were broadly distributed among the kochujang samples. Each sample contained a high abundance of region-specific bacterial species, such as B. sonorensis, B. pumilus, Weissella salipiscis, and diverse unidentified Bacillus species. Phylotype- and phylogeny-based community comparison analysis showed that the microbial communities of the two commercial brands were different from those of the local brands. Moreover, each local brand kochujang sample had region-specific microbial community reflecting the manufacturing environment. © 2012 Institute of Food Technologists®

Effect of different nickel precursors on capacitive behavior of electrodeposited NiO thin films

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

Kore, R. M.; Ghadge, T. S.; Ambare, R. C.

2016-04-13

In the present study, the effect of nickel precursors containing different anions like nitrate, chloride and sulphate on the morphology and pseudocapacitance behavior of NiO is investigated. The NiO samples were prepared by using a potentiondynamic electrodeposition technique in the three electrode cell. Cyclic voltammetry technique was exploited for potentiodynamic deposition of the films. The obtained samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), etc. The XRD reveals the cubic crystal structure for all samples. The SEM micrograph shows nanoflakelike, up grown nanoflakes and honeycomb like nanostructured morphologies for nitrate, chloride and sulphate precursors respectively. The capacitivemore » behavior of these samples was recorded using cyclic voltammetry (CV), charge-discharge and electrochemical impedance spectroscopy (EIS) in 1 M KOH electrolyte. The specific capacitance values of NiO samples obtained using CV for nitrate, chloride and sulphate precursors were 136, 214 and 893 Fg{sup −1} respectively, at the scan rate of 5 mVs{sup −1}. The charge discharge study shows high specific energy for the sample obtained from sulphate (23.98 Whkg{sup −1}) as compared to chloride (9.67 Whkg{sup −1}) and nitrate (4.9 Whkg{sup −1}), whereas samples of cholride (13.9 kWkg{sup −1} and nitrate (10.5 kWkg{sup −1}) shows comparatively more specific power than samples obtained from sulphate (7.6 kWkg{sup −1}). The equivalent series resistance of NiO samples observed from EIS study are 1.34, 1.29 and 1.27 Ω respectively for nitrate, chloride and sulphate precursors. These results emphasizes that the samples obtained from sulphate precursors provides very low impedance through honeycomb like nanostructured morphology which supports good capacitive behavior of NiO.« less

[Value of positive auto controls in the gel centrifugation method].

PubMed

Eichler, H; Kretschmer, V

1994-01-01

We studied 97 samples of patients being positive in the autocontrol of the indirect antiglobulin test (IAT) in the gel system (DiaMed). In 83.2%, retesting with monospecific anti-IgG serum gave also positive results, due to a specific phenomenon caused, for example, by drug-specific antibodies (AB), warm auto-AB or allo-AB. In contrast, only 52.9% of the samples retested by the standard tube technique with polyspecific antiglobulin serum reacted positive. Only in 6 patients slightly increased cold agglutinins could be detected. None of the investigated patients showed any clinical or laboratory signs of hemolysis except one with pernicious anemia. We conclude that positive results of the autocontrol in the gel IAT should be confirmed by an additional DAT in the tube technique. If this second test shows a negative result, transfusions can take place without any restrictions.

Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue.

PubMed

Miles, Timothy D; Martin, Frank N; Coffey, Michael D

2015-02-01

Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing, thereby reducing the time necessary for accurate diagnostics and making management decisions.

An Analysis of Nondestructive Evaluation Techniques for Polymer Matrix Composite Sandwich Materials

NASA Technical Reports Server (NTRS)

Cosgriff, Laura M.; Roberts, Gary D.; Binienda, Wieslaw K.; Zheng, Diahua; Averbeck, Timothy; Roth, Donald J.; Jeanneau, Philippe

2006-01-01

Structural sandwich materials composed of triaxially braided polymer matrix composite material face sheets sandwiching a foam core are being utilized for applications including aerospace components and recreational equipment. Since full scale components are being made from these sandwich materials, it is necessary to develop proper inspection practices for their manufacture and in-field use. Specifically, nondestructive evaluation (NDE) techniques need to be investigated for analysis of components made from these materials. Hockey blades made from sandwich materials and a flat sandwich sample were examined with multiple NDE techniques including thermographic, radiographic, and shearographic methods to investigate damage induced in the blades and flat panel components. Hockey blades used during actual play and a flat polymer matrix composite sandwich sample with damage inserted into the foam core were investigated with each technique. NDE images from the samples were presented and discussed. Structural elements within each blade were observed with radiographic imaging. Damaged regions and some structural elements of the hockey blades were identified with thermographic imaging. Structural elements, damaged regions, and other material variations were detected in the hockey blades with shearography. Each technique s advantages and disadvantages were considered in making recommendations for inspection of components made from these types of materials.

Screening for trace explosives by AccuTOF™-DART®: an in-depth validation study.

PubMed

Sisco, Edward; Dake, Jeffrey; Bridge, Candice

2013-10-10

Ambient ionization mass spectrometry is finding increasing utility as a rapid analysis technique in a number of fields. In forensic science specifically, analysis of many types of samples, including drugs, explosives, inks, bank dye, and lotions, has been shown to be possible using these techniques [1]. This paper focuses on one type of ambient ionization mass spectrometry, Direct Analysis in Real Time Mass Spectrometry (DART-MS or DART), and its viability as a screening tool for trace explosives analysis. In order to assess viability, a validation study was completed which focused on the analysis of trace amounts of nitro and peroxide based explosives. Topics which were studied, and are discussed, include method optimization, reproducibility, sensitivity, development of a search library, discrimination of mixtures, and blind sampling. Advantages and disadvantages of this technique over other similar screening techniques are also discussed. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Cryptosporidium Oocyst Detection in Water Samples: Floatation Technique Enhanced with Immunofluorescence Is as Effective as Immunomagnetic Separation Method

PubMed Central

Koompapong, Khuanchai; Sutthikornchai, Chantira

2009-01-01

Cryptosporidium can cause gastrointestinal diseases worldwide, consequently posing public health problems and economic burden. Effective techniques for detecting contaminated oocysts in water are important to prevent and control the contamination. Immunomagnetic separation (IMS) method has been widely employed recently due to its efficiency, but, it is costly. Sucrose floatation technique is generally used for separating organisms by using their different specific gravity. It is effective and cheap but time consuming as well as requiring highly skilled personnel. Water turbidity and parasite load in water sample are additional factors affecting to the recovery rate of those 2 methods. We compared the efficiency of IMS and sucrose floatation methods to recover the spiked Cryptosporidium oocysts in various turbidity water samples. Cryptosporidium oocysts concentration at 1, 101, 102, and 103 per 10 µl were spiked into 3 sets of 10 ml-water turbidity (5, 50, and 500 NTU). The recovery rate of the 2 methods was not different. Oocyst load at the concentration < 102 per 10 ml yielded unreliable results. Water turbidity at 500 NTU decreased the recovery rate of both techniques. The combination of sucrose floatation and immunofluorescense assay techniques (SF-FA) showed higher recovery rate than IMS and immunofluorescense assay (IMS-FA). We used this SF-FA to detect Cryptosporidium and Giardia from the river water samples and found 9 and 19 out of 30 (30% and 63.3%) positive, respectively. Our results favored sucrose floatation technique enhanced with immunofluorescense assay for detecting contaminated protozoa in water samples in general laboratories and in the real practical setting. PMID:19967082

Cryptosporidium oocyst detection in water samples: floatation technique enhanced with immunofluorescence is as effective as immunomagnetic separation method.

PubMed

Koompapong, Khuanchai; Sutthikornchai, Chantira; Sukthana, Yowalark

2009-12-01

Cryptosporidium can cause gastrointestinal diseases worldwide, consequently posing public health problems and economic burden. Effective techniques for detecting contaminated oocysts in water are important to prevent and control the contamination. Immunomagnetic separation (IMS) method has been widely employed recently due to its efficiency, but, it is costly. Sucrose floatation technique is generally used for separating organisms by using their different specific gravity. It is effective and cheap but time consuming as well as requiring highly skilled personnel. Water turbidity and parasite load in water sample are additional factors affecting to the recovery rate of those 2 methods. We compared the efficiency of IMS and sucrose floatation methods to recover the spiked Cryptosporidium oocysts in various turbidity water samples. Cryptosporidium oocysts concentration at 1, 10(1), 10(2), and 10(3) per 10 microl were spiked into 3 sets of 10 ml-water turbidity (5, 50, and 500 NTU). The recovery rate of the 2 methods was not different. Oocyst load at the concentration < 10(2) per 10 ml yielded unreliable results. Water turbidity at 500 NTU decreased the recovery rate of both techniques. The combination of sucrose floatation and immunofluorescense assay techniques (SF-FA) showed higher recovery rate than IMS and immunofluorescense assay (IMS-FA). We used this SF-FA to detect Cryptosporidium and Giardia from the river water samples and found 9 and 19 out of 30 (30% and 63.3%) positive, respectively. Our results favored sucrose floatation technique enhanced with immunofluorescense assay for detecting contaminated protozoa in water samples in general laboratories and in the real practical setting.

Task-specific singing dystonia: vocal instability that technique cannot fix.

PubMed

Halstead, Lucinda A; McBroom, Deanna M; Bonilha, Heather Shaw

2015-01-01

Singer's dystonia is a rare variation of focal laryngeal dystonia presenting only during specific tasks in the singing voice. It is underdiagnosed since it is commonly attributed to technique problems including increased muscle tension, register transition, or wobble. Singer's dystonia differs from technique-related issues in that it is task- and/or pitch-specific, reproducible and occurs independently from the previously mentioned technical issues.This case series compares and contrasts profiles of four patients with singer's dystonia to increase our knowledge of this disorder. This retrospective case series includes a detailed case history, results of singing evaluations from individual voice teachers, review of singing voice samples by a singing voice specialist, evaluation by a laryngologist with endoscopy and laryngeal electromyography (LEMG), and spectral analysis of the voice samples by a speech-language pathologist. Results demonstrate the similarities and unique differences of individuals with singer's dystonia. Response to treatment and singing status varied from nearly complete relief of symptoms with botulinum toxin injections to minor relief of symptoms and discontinuation of singing. The following are the conclusions from this case series: (1) singer's dystonia exists as a separate entity from technique issues, (2) singer's dystonia is consistent with other focal task-specific dystonias found in musicians, (3) correctly diagnosing singer's dystonia allows singer's access to medical treatment of dystonia and an opportunity to modify their singing repertoire to continue singing with the voice they have, and (4) diagnosis of singer's dystonia requires careful sequential multidisciplinary evaluation to isolate the instability and confirm dystonia by LEMG and spectral voice analysis. Copyright © 2015 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Comparative analysis of prostate-specific antigen by two-dimensional gel electrophoresis and capillary electrophoresis.

PubMed

Barrabés, Sílvia; Farina-Gomez, Noemi; Llop, Esther; Puerta, Angel; Diez-Masa, Jose Carlos; Perry, Antoinette; de Llorens, Rafael; de Frutos, Mercedes; Peracaula, Rosa

2017-02-01

Serum levels of Prostate-Specific Antigen (PSA) are not fully specific for prostate cancer (PCa) diagnosis and several efforts are focused on searching to improve PCa markers through the study of PSA subforms that could be cancer associated. We have previously reported by 2DE a decrease in the sialic acid content of PSA from PCa compared to benign prostatic hyperplasia patients based on the different proportion of the PSA spots. However, faster and more quantitative techniques, easier to automate than 2DE, are desirable. In this study, we examined the potential of CE for resolving PSA subforms in different samples and compared the results with those obtained by 2DE. We first fractionated by OFFGEL the subforms of PSA from seminal plasma according to their pIs and analyzed each separated fraction by 2DE and CE. We also analyzed PSA and high pI PSA, both from seminal plasma, and PSA from urine of a PCa patient. These samples with different PSA spots proportions by 2DE, due to different posttranslational modifications, also presented different CE profiles. This study shows that CE is a useful and complementary technique to 2DE for analyzing samples with different PSA subforms, which is of high clinical interest. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The Preliminary Examination of Organics in the Returned Stardust Samples from Comet Wild 2

NASA Technical Reports Server (NTRS)

Sandford, S. A.; Aleon, J.; Alexander, C.; Butterworth, A.; Clemett, S. J.; Cody, G.; Cooper, G.; Dworkin, J. P.; Flynn, G. J.; Gilles, M. K.

2006-01-01

The primary objective of STARDUST is to collect coma samples from comet 8lP/Wild 2. These samples were collected by impact onto aerogel tiles on Jan 2, 2004 when the spacecraft flew through the comet's coma at a relative velocity of about 6.1 km/sec. Measurements of dust impacts on the front of the spacecraft suggest that the aerogel particle collector was impacted by 2800 +/- 500 particles larger than 15 micron in diameter. Following recovery of the Sample Return Capsule (SRC) on Jan 15, 2006, the aerogel collector trays will be removed in a clean room at JSC. After documentation of the collection, selected aerogel tiles will be removed and aerogel and cometary samples will be extracted for study. A number of different extraction techniques will be used, each optimized for the analytical technique that is to be used. The STARDUST Mission will carry out a 6 month preliminary examination (PE) of a small portion of the returned samples. The examination of the samples will be made by a number of subteams that will concentrate on specific aspects of the samples. One of these is the Organics PE Team (see the author list above for team members). These team members will use a number of analytical techniques to produce a preliminary characterization of the abundance and nature of the organics (if any) in the returned samples.

ANALYSIS OF SAMPLING TECHNIQUES FOR IMBALANCED DATA: AN N=648 ADNI STUDY

PubMed Central

Dubey, Rashmi; Zhou, Jiayu; Wang, Yalin; Thompson, Paul M.; Ye, Jieping

2013-01-01

Many neuroimaging applications deal with imbalanced imaging data. For example, in Alzheimer’s Disease Neuroimaging Initiative (ADNI) dataset, the mild cognitive impairment (MCI) cases eligible for the study are nearly two times the Alzheimer’s disease (AD) patients for structural magnetic resonance imaging (MRI) modality and six times the control cases for proteomics modality. Constructing an accurate classifier from imbalanced data is a challenging task. Traditional classifiers that aim to maximize the overall prediction accuracy tend to classify all data into the majority class. In this paper, we study an ensemble system of feature selection and data sampling for the class imbalance problem. We systematically analyze various sampling techniques by examining the efficacy of different rates and types of undersampling, oversampling, and a combination of over and under sampling approaches. We thoroughly examine six widely used feature selection algorithms to identify significant biomarkers and thereby reduce the complexity of the data. The efficacy of the ensemble techniques is evaluated using two different classifiers including Random Forest and Support Vector Machines based on classification accuracy, area under the receiver operating characteristic curve (AUC), sensitivity, and specificity measures. Our extensive experimental results show that for various problem settings in ADNI, (1). a balanced training set obtained with K-Medoids technique based undersampling gives the best overall performance among different data sampling techniques and no sampling approach; and (2). sparse logistic regression with stability selection achieves competitive performance among various feature selection algorithms. Comprehensive experiments with various settings show that our proposed ensemble model of multiple undersampled datasets yields stable and promising results. PMID:24176869

Comparison of polymerase chain reaction and Warthin-Starry techniques to detect Leptospira spp. in kidneys of slaughtered cattle.

PubMed

Azizi, Shahrzad; Kheirandish, Reza; Rahimi, Elham

2014-11-12

Leptospirosis is a worldwide zoonotic disease that is caused by Gram-negative spirochaetes, Leptospira species. Affected animals excrete the organism in the urine into the environment and act as a source of infection. Cattle are maintenance hosts for some serovars of leptospirosis and are important in the transmission of the infection to humans. At post mortem examination, affected cattle show white spots in their kidneys but these are not specific for leptospirosis. Sometimes it is necessary that leptospirosis be diagnosed in the carcass. Different direct methods, including polymerase chain reaction (PCR), Warthin-Starry silver stain (WS), immunofluorescence (IF) and immunohistochemistry (IHC) can be used in order to diagnose leptospirosis in the affected tissues, such as kidney. The main advantage of the WS technique is direct visualisation of the bacteria in the tissue samples. Silver staining is useful for retrospective studies on formalin-fixed and paraffin-embedded samples but little information is available on the sensitivity and specificity of the technique. The present study aimed to find a simple and inexpensive method that can be used in any laboratory and that also, if clinical samples are not available, can detect Leptospira in tissue samples post mortem. This study was performed on 19 paraffin-embedded kidneys of slaughtered cows that grossly had focal to multifocal white spots. Leptospirosis was confirmed in these samples with PCR based on the LipL32 gene. Out of 19 PCR positive kidneys, Leptospira was identified in 13 stained samples by WS. The kidneys revealed different grades of interstitial nephritis. No relationship was found between severity of lesions and presence of leptospires in the kidneys. The PCR results on the urine and blood were consistent with matching WS stained kidneys. Out of 13 kidneys that were positive with silver staining, 7 matching blood and 10 matching urine samples were confirmed positive for leptospirosis with PCR. In this study, the WS technique provided fewer positive results than PCR. This may be as a result of a low burden of Leptospira in the kidney, but the sensitivity of WS staining needs more investigation.

Use of Tc-99m-galactosyl-neoglycoalbumin (Tc-NGA) to determine hepatic blood flow

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

Stadalnik, R.C.; Vera, D.R.; Woodle, E.S.

1984-01-01

Tc-NGA is a new liver radiopharmaceutical which binds to a hepatocyte-specific membrane receptor. Three characteristics of Tc-NGA can be exploited in the measurement of hepatic blood flow (HBF): 1) ability to alter the affinity of Tc-NGA for its receptor by changing the galactose: albumin ratio; 2) ability to achieve a high specific activity with Tc-99m labeling; and 3) ability to administer a high molar dose of Tc-NGA without physiologic side effects. In addition, kinetic modeling of Tc-NGA dynamic data can provide estimates of hepatic receptor concentration. In experimental studies in young pigs, HBF was determined using two techniques: 1) kineticmore » modeling of dynamic data using moderate affinity, low specific activity Tc-NGA (Group A, n=12); and 2) clearance (CL) technique using high affinity, high specific activity Tc-NGA (Group B, n=4). In both groups, HBF was determined simultaneously by continuous infusion of indocyanine green (CI-ICG) with hepatic vein sampling. Regression analysis of HBF measurements obtained with the Tc-NGA kinetic modeling technique and the CI-ICG technique (Group A) revealed good correlation between the two techniques (r=0.802, p=0.02). Similarly, HBF determination by the clearance technique (Group B) provided highly accurate measurements when compared to the CI-ICG technique. Hepatic blood flow measurements by the clearance technique (CL-NGA) fell within one standard deviation of the error associated with each CI-ICG HBF measurement (all CI-ICG standard deviations were less than 10%).« less

Method for concentration and separation of biological organisms by ultrafiltration and dielectrophoresis

DOEpatents

Simmons, Blake A.; Hill, Vincent R.; Fintschenko, Yolanda; Cummings, Eric B.

2012-09-04

Disclosed is a method for monitoring sources of public water supply for a variety of pathogens by using a combination of ultrafiltration techniques together dielectrophoretic separation techniques. Because water-borne pathogens, whether present due to "natural" contamination or intentional introduction, would likely be present in drinking water at low concentrations when samples are collected for monitoring or outbreak investigations, an approach is needed to quickly and efficiently concentrate and separate particles such as viruses, bacteria, and parasites in large volumes of water (e.g., 100 L or more) while simultaneously reducing the sample volume to levels sufficient for detecting low concentrations of microbes (e.g., <10 mL). The technique is also designed to screen the separated microbes based on specific conductivity and size.

The Microbial Detection Array Combined with Random Phi29-Amplification Used as a Diagnostic Tool for Virus Detection in Clinical Samples

PubMed Central

Erlandsson, Lena; Rosenstierne, Maiken W.; McLoughlin, Kevin; Jaing, Crystal; Fomsgaard, Anders

2011-01-01

A common technique used for sensitive and specific diagnostic virus detection in clinical samples is PCR that can identify one or several viruses in one assay. However, a diagnostic microarray containing probes for all human pathogens could replace hundreds of individual PCR-reactions and remove the need for a clear clinical hypothesis regarding a suspected pathogen. We have established such a diagnostic platform for random amplification and subsequent microarray identification of viral pathogens in clinical samples. We show that Phi29 polymerase-amplification of a diverse set of clinical samples generates enough viral material for successful identification by the Microbial Detection Array, demonstrating the potential of the microarray technique for broad-spectrum pathogen detection. We conclude that this method detects both DNA and RNA virus, present in the same sample, as well as differentiates between different virus subtypes. We propose this assay for diagnostic analysis of viruses in clinical samples. PMID:21853040

Comparison of Quantifiler(®) Trio and InnoQuant™ human DNA quantification kits for detection of DNA degradation in developed and aged fingerprints.

PubMed

Goecker, Zachary C; Swiontek, Stephen E; Lakhtakia, Akhlesh; Roy, Reena

2016-06-01

The development techniques employed to visualize fingerprints collected from crime scenes as well as post-development ageing may result in the degradation of the DNA present in low quantities in such evidence samples. Amplification of the DNA samples with short tandem repeat (STR) amplification kits may result in partial DNA profiles. A comparative study of two commercially available quantification kits, Quantifiler(®) Trio and InnoQuant™, was performed on latent fingerprint samples that were either (i) developed using one of three different techniques and then aged in ambient conditions or (ii) undeveloped and then aged in ambient conditions. The three fingerprint development techniques used were: cyanoacrylate fuming, dusting with black powder, and the columnar-thin-film (CTF) technique. In order to determine the differences between the expected quantities and actual quantities of DNA, manually degraded samples generated by controlled exposure of DNA standards to ultraviolet radiation were also analyzed. A total of 144 fingerprint and 42 manually degraded DNA samples were processed in this study. The results indicate that the InnoQuant™ kit is capable of producing higher degradation ratios compared to the Quantifiler(®) Trio kit. This was an expected result since the degradation ratio is a relative value specific for a kit based on the length and extent of amplification of the two amplicons that vary from one kit to the other. Additionally, samples with lower concentrations of DNA yielded non-linear relationships of degradation ratio with the duration of aging, whereas samples with higher concentrations of DNA yielded quasi-linear relationships. None of the three development techniques produced a noticeably different degradation pattern when compared to undeveloped fingerprints, and therefore do not impede downstream DNA analysis. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Accelerometer Data Analysis and Presentation Techniques

NASA Technical Reports Server (NTRS)

Rogers, Melissa J. B.; Hrovat, Kenneth; McPherson, Kevin; Moskowitz, Milton E.; Reckart, Timothy

1997-01-01

The NASA Lewis Research Center's Principal Investigator Microgravity Services project analyzes Orbital Acceleration Research Experiment and Space Acceleration Measurement System data for principal investigators of microgravity experiments. Principal investigators need a thorough understanding of data analysis techniques so that they can request appropriate analyses to best interpret accelerometer data. Accelerometer data sampling and filtering is introduced along with the related topics of resolution and aliasing. Specific information about the Orbital Acceleration Research Experiment and Space Acceleration Measurement System data sampling and filtering is given. Time domain data analysis techniques are discussed and example environment interpretations are made using plots of acceleration versus time, interval average acceleration versus time, interval root-mean-square acceleration versus time, trimmean acceleration versus time, quasi-steady three dimensional histograms, and prediction of quasi-steady levels at different locations. An introduction to Fourier transform theory and windowing is provided along with specific analysis techniques and data interpretations. The frequency domain analyses discussed are power spectral density versus frequency, cumulative root-mean-square acceleration versus frequency, root-mean-square acceleration versus frequency, one-third octave band root-mean-square acceleration versus frequency, and power spectral density versus frequency versus time (spectrogram). Instructions for accessing NASA Lewis Research Center accelerometer data and related information using the internet are provided.

Informed decision-making before changing to RDT: a comparison of microscopy, rapid diagnostic test and molecular techniques for the diagnosis and identification of malaria parasites in Kassala, eastern Sudan.

PubMed

Osman, Mamoun M M; Nour, Bakri Y M; Sedig, Mohamed F; De Bes, Laura; Babikir, Adil M; Mohamedani, Ahmed A; Mens, Petra F

2010-12-01

Rapid diagnostic tests (RDTs) are promoted for the diagnosis of malaria in many countries. The question arises whether laboratories where the current method of diagnosis is microscopy should also switch to RDT. This problem was studied in Kassala, Sudan where the issue of switching to RDT is under discussion. Two hundred and three blood samples were collected from febrile patients suspected of having malaria. These were subsequently analysed with microscopy, RDT (SD Bioline P.f/P.v) and PCR for the detection and identification of Plasmodium parasites. Malaria parasites were detected in 36 blood samples when examined microscopically, 54 (26.6%) samples were found positive for malaria parasites by RDT, and 44 samples were positive by PCR. Further analysis showed that the RDT used in our study resulted in a relatively high number of false positive samples. When microscopy was compared with PCR, an agreement of 96.1% and k = 0.88 (sensitivity 85.7% and specificity 100%) was found. However, when RDT was compared with PCR, an agreement of only 81.2 and k = 0.48 (sensitivity 69% and specificity 84%) was found. PCR has proven to be one of the most specific and sensitive diagnostic methods, particularly for malaria cases with low parasitaemia. However, this technique has limitations in its routine use under resource-limited conditions, such as our study location. At present, based on these results, microscopy remains the best option for routine diagnosis of malaria in Kassala, eastern Sudan. © 2010 Blackwell Publishing Ltd.

Applications of QCL mid-IR imaging to the advancement of pathology

NASA Astrophysics Data System (ADS)

Sreedhar, Hari; Varma, Vishal K.; Bird, Benjamin; Guzman, Grace; Walsh, Michael J.

2017-03-01

Quantum Cascade Laser (QCL) spectroscopic imaging is a novel technique with many potential applications to histopathology. Like traditional Fourier Transform Infrared (FT-IR) imaging, QCL spectroscopic imaging derives biochemical data coupled to the spatial information of a tissue sample, and can be used to improve the diagnostic and prognostic value of assessment of a tissue biopsy. This technique also offers advantages over traditional FT-IR imaging, specifically the capacity for discrete frequency and real-time imaging. In this work we present applications of QCL spectroscopic imaging to tissue samples, including discrete frequency imaging, to compare with FT-IR and its potential value to pathology.

Evaluation of Targeted Next-Generation Sequencing for Detection of Bovine Pathogens in Clinical Samples.

PubMed

Anis, Eman; Hawkins, Ian K; Ilha, Marcia R S; Woldemeskel, Moges W; Saliki, Jeremiah T; Wilkes, Rebecca P

2018-07-01

The laboratory diagnosis of infectious diseases, especially those caused by mixed infections, is challenging. Routinely, it requires submission of multiple samples to separate laboratories. Advances in next-generation sequencing (NGS) have provided the opportunity for development of a comprehensive method to identify infectious agents. This study describes the use of target-specific primers for PCR-mediated amplification with the NGS technology in which pathogen genomic regions of interest are enriched and selectively sequenced from clinical samples. In the study, 198 primers were designed to target 43 common bovine and small-ruminant bacterial, fungal, viral, and parasitic pathogens, and a bioinformatics tool was specifically constructed for the detection of targeted pathogens. The primers were confirmed to detect the intended pathogens by testing reference strains and isolates. The method was then validated using 60 clinical samples (including tissues, feces, and milk) that were also tested with other routine diagnostic techniques. The detection limits of the targeted NGS method were evaluated using 10 representative pathogens that were also tested by quantitative PCR (qPCR), and the NGS method was able to detect the organisms from samples with qPCR threshold cycle ( C T ) values in the 30s. The method was successful for the detection of multiple pathogens in the clinical samples, including some additional pathogens missed by the routine techniques because the specific tests needed for the particular organisms were not performed. The results demonstrate the feasibility of the approach and indicate that it is possible to incorporate NGS as a diagnostic tool in a cost-effective manner into a veterinary diagnostic laboratory. Copyright © 2018 Anis et al.

Detection of Hepatic Fibrosis in Ex Vivo Liver Samples Using an Open-Photoacoustic-Cell Method: Feasibility Study

NASA Astrophysics Data System (ADS)

Stolik, S.; Fabila, D. A.; de la Rosa, J. M.; Escobedo, G.; Suárez-Álvarez, K.; Tomás, S. A.

2015-09-01

Design of non-invasive and accurate novel methods for liver fibrosis diagnosis has gained growing interest. Different stages of liver fibrosis were induced in Wistar rats by intraperitoneally administering different doses of carbon tetrachloride. The liver fibrosis degree was conventionally determined by means of histological examination. An open-photoacoustic-cell (OPC) technique for the assessment of liver fibrosis was developed and is reported here. The OPC technique is based on the fact that the thermal diffusivity can be accurately measured by photoacoustics taking into consideration the photoacoustic signal amplitude versus the modulation frequency. This technique measures directly the heat generated in a sample, due to non-radiative de-excitation processes, following the absorption of light. The thermal diffusivity was measured with a home-made open-photoacoustic-cell system that was specially designed to perform the measurement from ex vivo liver samples. The human liver tissue showed a significant increase in the thermal diffusivity depending on the fibrosis stage. Specifically, liver samples from rats exhibiting hepatic fibrosis showed a significantly higher value of the thermal diffusivity than for control animals.

Orthogonality-breaking sensing model based on the instantaneous Stokes vector and the Mueller calculus

NASA Astrophysics Data System (ADS)

Ortega-Quijano, Noé; Fade, Julien; Roche, Muriel; Parnet, François; Alouini, Mehdi

2016-04-01

Polarimetric sensing by orthogonality breaking has been recently proposed as an alternative technique for performing direct and fast polarimetric measurements using a specific dual-frequency dual-polarization (DFDP) source. Based on the instantaneous Stokes-Mueller formalism to describe the high-frequency evolution of the DFDP beam intensity, we thoroughly analyze the interaction of such a beam with birefringent, dichroic and depolarizing samples. This allows us to confirm that orthogonality breaking is produced by the sample diattenuation, whereas this technique is immune to both birefringence and diagonal depolarization. We further analyze the robustness of this technique when polarimetric sensing is performed through a birefringent waveguide, and the optimal DFDP source configuration for fiber-based endoscopic measurements is subsequently identified. Finally, we consider a stochastic depolarization model based on an ensemble of random linear diattenuators, which makes it possible to understand the progressive vanishing of the detected orthogonality breaking signal as the spatial heterogeneity of the sample increases, thus confirming the insensitivity of this method to diagonal depolarization. The fact that the orthogonality breaking signal is exclusively due to the sample dichroism is an advantageous feature for the precise decoupled characterization of such an anisotropic parameter in samples showing several simultaneous effects.

Module Degradation Mechanisms Studied by a Multi-Scale Approach

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

Johnston, Steve; Al-Jassim, Mowafak; Hacke, Peter

2016-11-21

A key pathway to meeting the Department of Energy SunShot 2020 goals is to reduce financing costs by improving investor confidence through improved photovoltaic (PV) module reliability. A comprehensive approach to further understand and improve PV reliability includes characterization techniques and modeling from module to atomic scale. Imaging techniques, which include photoluminescence, electroluminescence, and lock-in thermography, are used to locate localized defects responsible for module degradation. Small area samples containing such defects are prepared using coring techniques and are then suitable and available for microscopic study and specific defect modeling and analysis.

Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs.

PubMed

Borecka, A; Gawor, J

2008-06-01

A modification of gDNA extraction was developed for the polymerase chain reaction (PCR) technique, intended for the detection and differentiation of Toxocara spp. eggs in soil or sediments. Sand samples from sandpits confirmed as being contaminated with Toxocara spp. eggs by the flotation technique were analysed by PCR. The use of proteinase K made it possible to obtain genomic DNA from the sample without needing to isolate eggs using flotation or to inactivate PCR inhibitors present in the sand. Specific primers in the PCR reaction allowed discrimination between T. canis and T. cati eggs. The modification simplified the procedure, thanks to eliminating the step of gDNA isolation from eggs, which is both laborious and difficult.

DETECTION OF PATHOGENS IN DRINKING WATER (SEER 2)

EPA Science Inventory

Project investigators developed a polymerase chain reaction (PCR)-based technique to detect E. coli 0157:H7 cells in environmental samples using previously reported PCR primers for the specific detection of genes involved in biosynthesis of 0157 polysacchari...

Application of Faecalibacterium 16S rDNA genetic marker for accurate identification of duck faeces.

PubMed

Sun, Da; Duan, Chuanren; Shang, Yaning; Ma, Yunxia; Tan, Lili; Zhai, Jun; Gao, Xu; Guo, Jingsong; Wang, Guixue

2016-04-01

The aim of this study was to judge the legal duty of pollution liabilities by assessing a duck faeces-specific marker, which can exclude distractions of residual bacteria from earlier contamination accidents. With the gene sequencing technology and bioinformatics method, we completed the comparative analysis of Faecalibacterium sequences, which were associated with ducks and other animal species, and found the sequences unique to duck faeces. Polymerase chain reaction (PCR) and agarose gel electrophoresis techniques were used to verify the reliability of both human and duck faeces-specific primers. The duck faeces-specific primers generated an amplicon of 141 bp from 43.3 % of duck faecal samples, 0 % of control samples and 100 % of sewage wastewater samples that contained duck faeces. We present here the initial evidence of Faecalibacterium-based applicability as human faeces-specificity in China. Meanwhile, this study represents the initial report of a Faecalibacterium marker for duck faeces and suggests an independent or supplementary environmental biotechnology of microbial source tracking (MST).

High-throughput detection of human papillomavirus-18 L1 gene methylation, a candidate biomarker for the progression of cervical neoplasia.

PubMed

Turan, Tolga; Kalantari, Mina; Cuschieri, Kate; Cubie, Heather A; Skomedal, Hanne; Bernard, Hans-Ulrich

2007-04-25

The L1 gene of human papillomavirus-18 (HPV-18) is consistently hypermethylated in cervical carcinomas, but frequently hypo- or unmethylated in exfoliated cells from asymptomatic patients. In precancerous lesions, L1 is sporadically hypermethylated, correlating with the severity of the neoplasia. In order to explore the potential of using L1 methylation as a workable biomarker for carcinogenic progression of HPV-18 infections in routinely taken samples, our aim was to develop methylation-detection techniques that were sensitive and rapid without being overly complex technically. Therein, we developed a methylation-specific PCR (MSP) through the design of primer sets that specifically amplify either methylated or unmethylated HPV-18 L1 DNA within bisulfite-modified sample DNA. Amplification of unmethylated and in vitro methylated HPV-18 DNA by MSP resulted in 2500 copies of either of the two L1 DNA species being detected, a satisfactory sensitivity considering that bisulfite treatment leads to the fragmentation of about 99% of sample DNA. The primers proved specific and did not generate false positive results at concentrations exceeding the lowest limit of detection by a factor of 400. DNA from carcinomas yielded PCR signals only with the methylation-specific primers, and not with primers specific for unmethylated L1 genes. The inverse result was obtained with DNA from precursor lesions that contained only hypomethylated DNA. High-grade precursor lesions and carcinomas that contained hyper- as well as hypomethylated L1 DNA yielded PCR signals with both primers. By developing a fluorescence based real-time PCR, we quantitatively analyzed samples with in vitro methylated and unmethylated L1 DNA, and could distinguish clinical samples with hyper- and hypomethylated DNA or mixtures of both DNAs. The methylation-specific and real-time PCR techniques permitted efficient HPV-18 L1 methylation analyses and open the door for larger-scale clinical studies where the utility of methylation status to predict the progression of HPV-18 infection and HPV-18 associated lesions is assessed.

Biomechanical advantages of robot-assisted pedicle screw fixation in posterior lumbar interbody fusion compared with freehand technique in a prospective randomized controlled trial-perspective for patient-specific finite element analysis.

PubMed

Kim, Ho-Joong; Kang, Kyoung-Tak; Park, Sung-Cheol; Kwon, Oh-Hyo; Son, Juhyun; Chang, Bong-Soon; Lee, Choon-Ki; Yeom, Jin S; Lenke, Lawrence G

2017-05-01

There have been conflicting results on the surgical outcome of lumbar fusion surgery using two different techniques: robot-assisted pedicle screw fixation and conventional freehand technique. In addition, there have been no studies about the biomechanical issues between both techniques. This study aimed to investigate the biomechanical properties in terms of stress at adjacent segments using robot-assisted pedicle screw insertion technique (robot-assisted, minimally invasive posterior lumbar interbody fusion, Rom-PLIF) and freehand technique (conventional, freehand, open approach, posterior lumbar interbody fusion, Cop-PLIF) for instrumented lumbar fusion surgery. This is an additional post-hoc analysis for patient-specific finite element (FE) model. The sample is composed of patients with degenerative lumbar disease. Intradiscal pressure and facet contact force are the outcome measures. Patients were randomly assigned to undergo an instrumented PLIF procedure using a Rom-PLIF (37 patients) or a Cop-PLIF (41), respectively. Five patients in each group were selected using a simple random sampling method after operation, and 10 preoperative and postoperative lumbar spines were modeled from preoperative high-resolution computed tomography of 10 patients using the same method for a validated lumbar spine model. Under four pure moments of 7.5 Nm, the changes in intradiscal pressure and facet joint contact force at the proximal adjacent segment following fusion surgery were analyzed and compared with preoperative states. The representativeness of random samples was verified. Both groups showed significant increases in postoperative intradiscal pressure at the proximal adjacent segment under four moments, compared with the preoperative state. The Cop-PLIF models demonstrated significantly higher percent increments of intradiscal pressure at proximal adjacent segments under extension, lateral bending, and torsion moments than the Rom-PLIF models (p=.032, p=.008, and p=.016, respectively). Furthermore, the percent increment of facet contact force was significantly higher in the Cop-PLIF models under extension and torsion moments than in the Rom-PLIF models (p=.016 under both extension and torsion moments). The present study showed the clinical application of subject-specific FE analysis in the spine. Even though there was biomechanical superiority of the robot-assisted insertions in terms of alleviation of stress increments at adjacent segments after fusion, cautious interpretation is needed because of the small sample size. Copyright © 2016 Elsevier Inc. All rights reserved.

Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

PubMed Central

Ryu, Hodon; Henson, Michael; Elk, Michael; Toledo-Hernandez, Carlos; Griffith, John; Blackwood, Denene; Noble, Rachel; Gourmelon, Michèle; Glassmeyer, Susan

2013-01-01

The detection of environmental enterococci has been determined primarily by using culture-based techniques that might exclude some enterococcal species as well as those that are nonculturable. To address this, the relative abundances of enterococci were examined by challenging fecal and water samples against a currently available genus-specific assay (Entero1). To determine the diversity of enterococcal species, 16S rRNA gene-based group-specific quantitative PCR (qPCR) assays were developed and evaluated against eight of the most common environmental enterococcal species. Partial 16S rRNA gene sequences of 439 presumptive environmental enterococcal strains were analyzed to study further the diversity of enterococci and to confirm the specificities of group-specific assays. The group-specific qPCR assays showed relatively high amplification rates with targeted species (>98%), although some assays cross-amplified with nontargeted species (1.3 to 6.5%). The results with the group-specific assays also showed that different enterococcal species co-occurred in most fecal samples. The most abundant enterococci in water and fecal samples were Enterococcus faecalis and Enterococcus faecium, although we identified more water isolates as Enterococcus casseliflavus than as any of the other species. The prevalence of the Entero1 marker was in agreement with the combined number of positive signals determined by the group-specific assays in most fecal samples, except in gull feces. On the other hand, the number of group-specific assay signals was lower in all water samples tested, suggesting that other enterococcal species are present in these samples. While the results highlight the value of genus- and group-specific assays for detecting the major enterococcal groups in environmental water samples, additional studies are needed to determine further the diversity, distributions, and relative abundances of all enterococcal species found in water. PMID:23087032

[Molecular identification of Hibiscus syriacus and its adulterants using ITS2 barcode].

PubMed

Liu, Yi-Mei; Jin, Li-Na; Xiong, Yong-Xin; Wu, Lan; Chen, Ke-Li

2014-03-01

To identify Hibiscus syriacus and its adulterants using DNA barcoding technique. Nine samples of five species were PCR amplified and sequenced, and twelve samples were downloaded from the GenBank. The intra-specific and interspecific K2P distances were calculated, and neighbor-joining( NJ) tree was constructed by MEGA 5.0. The results showed the intra-specific genetic distances of Hibiscus syriacus were ranged from 0.009 to 0.056, which were far lower than inter-specific genetic distances between Hibiscus syriacus and its adulterants (0.236 - 0.301). Variable sites within Hibiscus syriacus ranged from 2 to 9 which were far less than the adulterants (45 - 52); Different samples of Hibiscus syriacus were gathered together and could be distinguished from its adulterants by NJ tree. ITS2 can discriminate Hibiscus syriacus from its adulterants correctly. The ITS2 region is an efficient barcode for authentication of Hibiscus syriacus and its adulterants.

Application of Synchrotron Microprobe Methods to Solid-Phase Speciation of Metals and Metalloids in House Dust

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

S Walker; H Jamieson; P Rasmussen

2011-12-31

Determination of the source and form of metals in house dust is important to those working to understand human and particularly childhood exposure to metals in residential environments. We report the development of a synchrotron microprobe technique for characterization of multiple metal hosts in house dust. We have applied X-ray fluorescence for chemical characterization and X-ray diffraction for crystal structure identification using microfocused synchrotron X-rays at a less than 10 {micro}m spot size. The technique has been evaluated by application to archived house dust samples containing elevated concentrations of Pb, Zn, and Ba in bedroom dust, and Pb and Asmore » in living room dust. The technique was also applied to a sample of soil from the corresponding garden to identify linkages between indoor and outdoor sources of metals. Paint pigments including white lead (hydrocerussite) and lithopone (wurtzite and barite) are the primary source of Pb, Zn, and Ba in bedroom dust, probably related to renovation activity in the home at the time of sampling. The much lower Pb content in the living room dust shows a relationship to the exterior soil and no specific evidence of Pb and Zn from the bedroom paint pigments. The technique was also successful at confirming the presence of chromated copper arsenate treated wood as a source of As in the living room dust. The results of the study have confirmed the utility of this approach in identifying specific metal forms within the dust.« less

Review of Fluorescence-Based Velocimetry Techniques to Study High-Speed Compressible Flows

NASA Technical Reports Server (NTRS)

Bathel, Brett F.; Johansen, Criag; Inman, Jennifer A.; Jones, Stephen B.; Danehy, Paul M.

2013-01-01

This paper reviews five laser-induced fluorescence-based velocimetry techniques that have been used to study high-speed compressible flows at NASA Langley Research Center. The techniques discussed in this paper include nitric oxide (NO) molecular tagging velocimetry (MTV), nitrogen dioxide photodissociation (NO2-to-NO) MTV, and NO and atomic oxygen (O-atom) Doppler-shift-based velocimetry. Measurements of both single-component and two-component velocity have been performed using these techniques. This paper details the specific application and experiment for which each technique has been used, the facility in which the experiment was performed, the experimental setup, sample results, and a discussion of the lessons learned from each experiment.

SIMSISH Technique Does Not Alter the Apparent Isotopic Composition of Bacterial Cells

PubMed Central

Chapleur, Olivier; Wu, Ting-Di; Guerquin-Kern, Jean-Luc; Mazéas, Laurent; Bouchez, Théodore

2013-01-01

In order to identify the function of uncultured microorganisms in their environment, the SIMSISH method, combining in situ hybridization (ISH) and nanoscale secondary ion mass spectrometry (nanoSIMS) imaging, has been proposed to determine the quantitative uptake of specific labelled substrates by uncultured microbes at the single cell level. This technique requires the hybridization of rRNA targeted halogenated DNA probes on fixed and permeabilized microorganisms. Exogenous atoms are introduced into cells and endogenous atoms removed during the experimental procedures. Consequently differences between the original and the apparent isotopic composition of cells may occur. In the present study, the influence of the experimental procedures of SIMSISH on the isotopic composition of carbon in E. coli cells was evaluated with nanoSIMS and compared to elemental analyser-isotopic ratio mass spectrometer (EA-IRMS) measurements. Our results show that fixation and hybridization have a very limited, reproducible and homogeneous influence on the isotopic composition of cells. Thereby, the SIMSISH procedure minimizes the contamination of the sample by exogenous atoms, thus providing a means to detect the phylogenetic identity and to measure precisely the carbon isotopic composition at the single cell level. This technique was successfully applied to a complex sample with double bromine – iodine labelling targeting a large group of bacteria and a specific archaea to evaluate their specific 13C uptake during labelled methanol anaerobic degradation. PMID:24204855

Molecular diagnosis of rhino-orbito-cerebral mucormycosis from fresh tissue samples.

PubMed

Zaman, Kamran; Rudramurthy, Shivaprakash Mandya; Das, Ashim; Panda, Naresh; Honnavar, Prasanna; Kaur, Harsimran; Chakrabarti, Arunaloke

2017-08-01

We aimed to evaluate a PCR-based technique for the diagnosis of mucormycosis and the identification of fungi from fresh tissue specimens in patients with rhino-orbito-cerebral-mucormycosis (ROCM). Fifty cases of ROCM were included in the study. Conventional identification was performed using microscopy and culture. Molecular diagnosis was performed by amplifying the ribosomal DNA using pan-fungal ITS primers and semi-nested Mucorales-specific primers of the 18S region. The amplified products were sequenced to identify the agents. The utility of PCR-RFLP of the 18S region of rDNA was evaluated to identify the Mucorales. The ROCM cases were diagnosed by the demonstration of aseptate ribbon-like hyphae in biopsy specimens collected from the patients. Isolation was possible in 24 (48 %) samples. The ITS2 PCR confirmed mucormycosis in 27 cases (54 %; CI 59.4-68.2). By comparison, Mucorales-specific PCR was able to amplify DNA and the sequence enabled the identification of Mucorales speciesin all the patients. PCR-RFLP of the 18S region of rDNA could only identify the agent to genus level. The molecular technique was able to identify Mucorales species in 26 (42 %) cases that were negative by culture. Mucorales-specific semi-nested PCR targeting the 18S region is a better technique than ITS2 PCR for diagnosis. PCR-RFLP of the 18S region helps in identification to genus level.

A geochemical sampling technique for use in areas of active alpine glaciation: an application from the central Alaska Range

USGS Publications Warehouse

Stephens, G.C.; Evenson, E.B.; Detra, D.E.

1990-01-01

In mountainous regions containing extensive glacier systems there is a lack of suitable material for conventional geochemical sampling. As a result, in most geochemical sampling programs a few stream-sediment samples collected at, or near, the terminus of valley glaciers are used to evaluate the mineral potential of the glaciated area. We have developed and tested a technique which utilizes the medial moraines of valley glaciers for systematic geochemical exploration of the glacial catchment area. Moraine sampling provides geochemical information that is site-specific in that geochemical anomalies can be traced directly up-ice to bedrock sources. Traverses were made across the Trident and Susitna glaciers in the central Alaska Range where fine-grained (clay to sand size) samples were collected from each medial moraine. These samples were prepared and chemically analyzed to determine the concentration of specific elements. Fifty pebbles were collected at each moraine for archival purposes and for subsequent lithologic identification. Additionally, fifty cobbles and fifty boulders were examined and described at each sample site to determine the nature and abundance of lithologies present in the catchment area, the extent and nature of visible mineralization, the presence and intensity of hydrothermal alteration and the existence of veins, dikes and other minor structural features. Results from the central Alaska Range have delineated four distinct multi-element anomalies which are a response to potential mineralization up-ice from the medial moraine traverse. By integrating the lithologic, mineralogical and geochemical data the probable geological setting of the geochemical anomalies is determined. ?? 1990.

Detection, characterization and quantification of inorganic engineered nanomaterials: A review of techniques and methodological approaches for the analysis of complex samples.

PubMed

Laborda, Francisco; Bolea, Eduardo; Cepriá, Gemma; Gómez, María T; Jiménez, María S; Pérez-Arantegui, Josefina; Castillo, Juan R

2016-01-21

The increasing demand of analytical information related to inorganic engineered nanomaterials requires the adaptation of existing techniques and methods, or the development of new ones. The challenge for the analytical sciences has been to consider the nanoparticles as a new sort of analytes, involving both chemical (composition, mass and number concentration) and physical information (e.g. size, shape, aggregation). Moreover, information about the species derived from the nanoparticles themselves and their transformations must also be supplied. Whereas techniques commonly used for nanoparticle characterization, such as light scattering techniques, show serious limitations when applied to complex samples, other well-established techniques, like electron microscopy and atomic spectrometry, can provide useful information in most cases. Furthermore, separation techniques, including flow field flow fractionation, capillary electrophoresis and hydrodynamic chromatography, are moving to the nano domain, mostly hyphenated to inductively coupled plasma mass spectrometry as element specific detector. Emerging techniques based on the detection of single nanoparticles by using ICP-MS, but also coulometry, are in their way to gain a position. Chemical sensors selective to nanoparticles are in their early stages, but they are very promising considering their portability and simplicity. Although the field is in continuous evolution, at this moment it is moving from proofs-of-concept in simple matrices to methods dealing with matrices of higher complexity and relevant analyte concentrations. To achieve this goal, sample preparation methods are essential to manage such complex situations. Apart from size fractionation methods, matrix digestion, extraction and concentration methods capable of preserving the nature of the nanoparticles are being developed. This review presents and discusses the state-of-the-art analytical techniques and sample preparation methods suitable for dealing with complex samples. Single- and multi-method approaches applied to solve the nanometrological challenges posed by a variety of stakeholders are also presented. Copyright © 2015 Elsevier B.V. All rights reserved.

Diagnostic value of JC/BK virus antibody immunohistochemistry staining in urine samples from posttransplant immunosuppressed patients in relation to polyomavirus reactivation.

PubMed

Yuste, Rosario Sanchez; Frías, Carolina; López, Ana; Vallejo, Carlos; Martín, Paloma; Bellas, Carmen

2008-01-01

To compare the diagnostic value of cytology and immunohistochemistry staining (IHS) of urine samples for polyomavirus reactivation diagnosis. Sixty-eight urine samples collected from 18 immunosuppressed patients were analyzed by Papanicolaou and IHS with a JC/BK virus-specific monoclonal antibody. Overall, polyomavirus BK (BKV) was positive in 11 of 18 patients (61.1%) (3 of whom developed hemorrhagic cystitis) and in 23 of 68 urine samples (28%). Of 23 samples, 4 (17%) were positive by 1 of the 2 techniques, only. Of 23 samples, 19 (83%) were positive by both methods. In matching urine samples from the same patient, the number of BKV-infected positive cells detected by IHS in urine slides was higher than those detected by Papanicolaou staining (71.3%). The main advantage of LHS is that it allowed confirmation of BKV infection diagnosis in urine samples. IHS detected more BKV-infected cells in samples with few positive urothelial cells, which would have gone undetected if only Papanicolaou staining had been used as the BKV screening method. Urine samples testing for BKV by both techniques will improve diagnosis in asymptomatic patients, allowing early therapeutic intervention and a better clinical outcome.

Experimental technique for studying high-temperature phase equilibria in reactive molten metal based systems

NASA Astrophysics Data System (ADS)

Ermoline, Alexandre

The general objective of this work is to develop an experimental technique for studying the high-temperature phase compositions and phase equilibria in molten metal-based binary and ternary systems, such as Zr-O-N, B-N-O, Al-O, and others. A specific material system of Zr-O-N was selected for studying and testing this technique. The information about the high-temperature phase equilibria in reactive metal-based systems is scarce and their studying is difficult because of chemical reactions occurring between samples and essentially any container materials, and causing contamination of the system. Containerless microgravity experiments for studying equilibria in molten metal-gas systems were designed to be conducted onboard of a NASA KC-135 aircraft flying parabolic trajectories. A uniaxial apparatus suitable for acoustic levitation, laser heating, and splat quenching of small samples was developed and equipped with computer-based controller and optical diagnostics. Normal-gravity tests were conducted to determine the most suitable operating parameters of the levitator by direct observations of the levitated samples, as opposed to more traditional pressure mapping of the acoustic field. The size range of samples that could be reliably heated and quenched in this setup was determined to be on the order of 1--3 mm. In microgravity experiments, small spherical specimens (1--2 mm diameter), prepared as pressed, premixed solid components, ZrO2, ZrN, and Zr powders, were acoustically levitated inside an argon-filled chamber at one atmosphere and heated by a CO2 laser. The levitating samples could be continuously laser heated for about 1 sec, resulting in local sample melting. The sample stability in the vertical direction was undisturbed by simultaneous laser heating. Oscillations of the levitating sample in the horizontal direction increased while it was heated, which eventually resulted in the movement of the sample away from its stable levitation position and the laser beam. The follow-up on-ground experiments were conducted to study phase relations in the Zr-O-N system at high-temperatures. Samples with specific compositions were laser-heated above the melt formation and naturally cooled. Recovered samples were characterized using electron microscopy, energy-dispersive spectroscopy, and x-ray diffraction. Results of these analyses combined with the interpretations of the binary Zr-O and Zr-N phase diagrams enabled us to outline the liquidus and the subsolidus equilibria for the ternary Zr-ZrO2-ZrN phase diagrams. Further research is suggested to develop the microgravity techniques for detailed characterization of high-temperature relations in the reactive, metal based systems.

Immunodiagnosis of fascioliasis using sandwich enzyme-linked immunosorbent assay for detection of Fasciola gigantica paramyosin antigen

PubMed Central

Abou-Elhakam, Hany Mohamed Adel; Bauomy, Ibraheem Rabia; El Deeb, Somaya Osman; El Amir, Azza Mohamed

2013-01-01

Background: Many immunological techniques have been developed over years using the different Fasciola antigens for diagnosis of parasitic infection and to replace the parasitological techniques, which are time consuming and usually lack sensitivity and reproducibility. Materials and Methods: In this study, Fasciola gigantica paramyosin (Pmy) antigen was early detected in cattle sera using sandwich enzyme-linked immunosorbent assay (ELISA), to evaluate the Pmy antigen performance in diagnosis. This work was conducted on 135 cattle blood samples, which were classified according to parasitological investigation into, healthy control (30), fascioliasis (75), and other parasites (30) groups. Results: The sensitivity of Sandwich ELISA was 97.33%, and the specificity was 95%, in comparison with parasitological examination, which recorded 66.66% sensitivity and 100% specificity, respectively. Conclusions: It was clear that the native F. gigantica Pmy is considered as a powerful antigen in early immunodiagnosis of fascioliasis, using a highly sensitive and specific sandwich ELISA technique. PMID:23961441

Regulated bioluminescence as a tool for bioremediation process monitoring and control of bacterial cultures

NASA Technical Reports Server (NTRS)

Burlage, Robert S.; Heitzer, Armin; Digrazia, Philip M.

1991-01-01

An effective on-line monitoring technique for toxic waste bioremediation using bioluminescent microorganisms has shown great potential for the description and optimization of biological processes. The lux genes of the bacterium Vibrio fischeri are used by this species to produce visible light. The lux genes can be genetically fused to the control region of a catabolic gene, with the result that bioluminescence is produced whenever the catabolic gene is induced. Thus the detection of light from a sample indicates that genetic expression from a specific gene is occurring. This technique was used to monitor biodegradation of specific contaminants from waste sites. For these studies, fusions between the lux genes and the operons for naphthalene and toluene/xylene degradation were constructed. Strains carrying one of these fusions respond sensitively and specifically to target substrates. Bioluminescence from these cultures can be rapidly measured in a nondestructive and noninvasive manner. The potential for this technique in this and other biological systems is discussed.

Composite technique for regional neurochemical studies: measurement of energy and neurotransmitter metabolites in single tissue sample.

PubMed

Djuricic, B M; Ueki, Y; Spatz, M

1985-06-01

A combined method is described for the determination of various metabolites from a single tissue sample of the brain. It comprises a quick inactivation of cerebral enzymes by microwave irradiation, easy separation of the desired brain regions, and perchloric acid extraction of tissue substances, which are assayed either by specific enzymatic techniques or by HPLC with electrochemical detection. The obtained values of most energy and neurotransmitter metabolites in the brain are in agreement with those reported using other methods. However, this technique, in contrast to the brain freezing in vitro or freeze-blowing, provides a more efficient procedure for rapid arrest of cerebral metabolism even in the deep brain structures and is therefore suitable for detection of early changes particularly those occurring in experimental pathological conditions such as ischemia.

Continuous gradient temperature Raman spectroscopy of unsaturated fatty acids

USDA-ARS?s Scientific Manuscript database

A new innovative technique gradient temperature, Raman spectroscopy (GTRS), identifies Raman frequency shifts in solid or liquid samples, and correlates them with specific temperature ranges within which flexible structures absorb heat. GTRS can easily detect changes that occur within one celcius te...

Utilizing the Zero-One Linear Programming Constraints to Draw Multiple Sets of Matched Samples from a Non-Treatment Population as Control Groups for the Quasi-Experimental Design

ERIC Educational Resources Information Center

Li, Yuan H.; Yang, Yu N.; Tompkins, Leroy J.; Modarresi, Shahpar

2005-01-01

The statistical technique, "Zero-One Linear Programming," that has successfully been used to create multiple tests with similar characteristics (e.g., item difficulties, test information and test specifications) in the area of educational measurement, was deemed to be a suitable method for creating multiple sets of matched samples to be…

Bladder cancer diagnosis during cystoscopy using Raman spectroscopy

NASA Astrophysics Data System (ADS)

Grimbergen, M. C. M.; van Swol, C. F. P.; Draga, R. O. P.; van Diest, P.; Verdaasdonk, R. M.; Stone, N.; Bosch, J. H. L. R.

2009-02-01

Raman spectroscopy is an optical technique that can be used to obtain specific molecular information of biological tissues. It has been used successfully to differentiate normal and pre-malignant tissue in many organs. The goal of this study is to determine the possibility to distinguish normal tissue from bladder cancer using this system. The endoscopic Raman system consists of a 6 Fr endoscopic probe connected to a 785nm diode laser and a spectral recording system. A total of 107 tissue samples were obtained from 54 patients with known bladder cancer during transurethral tumor resection. Immediately after surgical removal the samples were placed under the Raman probe and spectra were collected and stored for further analysis. The collected spectra were analyzed using multivariate statistical methods. In total 2949 Raman spectra were recorded ex vivo from cold cup biopsy samples with 2 seconds integration time. A multivariate algorithm allowed differentiation of normal and malignant tissue with a sensitivity and specificity of 78,5% and 78,9% respectively. The results show the possibility of discerning normal from malignant bladder tissue by means of Raman spectroscopy using a small fiber based system. Despite the low number of samples the results indicate that it might be possible to use this technique to grade identified bladder wall lesions during endoscopy.

Effect of polyaniline on MWCNTs supercapacitor properties prepared by electrophoretic deposition

NASA Astrophysics Data System (ADS)

Razak, Rozelia Azila Abd; Eleas, Nor Hamizah; Mohammad, Nurul Nazwa; Yusof, Azmi Mohamed; Zaine, Intan Syaffinazzilla

2017-08-01

Multi-walled carbon nanotubes (MWCNTs) is widely used as supercapacitor electrode material. However, the specific capacitance of MWCNTs cannot achieve optimum value to facilitate required demand. Conducting polymers have been introduced to achieve optimum energy density and power density of supercapacitor electrode material. Previous work had demonstrated the effects of adding conducting polymer into carbon base material to get pseudocapacitance effect. Nevertheless the effects specifically of polyaniline (PANi) to MWCNTs were significantly low. This work describes the effect of PANi adding on MWCNTs film prepared by electrophoretic deposition (EPD) technique in order to increase the specific capacitance of MWCNTs. The commercial MWCNTs is dispersed in deionized water by using crystal violet. The admixtures without PANi (sample A), 5wt.% of PANi (sample B) and 10wt.% of PANi (sample C) have been prepared by ex-situ polymerization. The voltage supplied for film deposition is 8 V for 5 minutes. The morphology, functional group and electrochemical properties of MWCNTs due to the presence of PANi had been studied. From FESEM analysis, the presence of PANi can be clearly observed for sample B and sample C while FTIR analysis, proves PANi structure on MWCNTs with its functional group presence in sample B and sample C through the absorbtion band which obviously shifted to higher value compare to sample A. Cyclic voltammogram (CV) analysis shown redox activity occurred in sample B and sample C with identical anodic and cathodic peaks. Sample B hold the higher specific capacitance and higher energy density compared than sample A and sample B. From galvanostatic charge-discharge (CD) measurement, the charge and discharge process for sample B is longer than sample A and sample C which consequently lower its power density. The presence of PANi at 5wt.% is able to increase specific capacitance as well as energy density to optimum value.

Development and application of mass spectrometric techniques for ultra-trace determination of 236U in environmental samples-A review.

PubMed

Bu, Wenting; Zheng, Jian; Ketterer, Michael E; Hu, Sheng; Uchida, Shigeo; Wang, Xiaolin

2017-12-01

Measurements of the long-lived radionuclide 236 U are an important endeavor, not only in nuclear safeguards work, but also in terms of using this emerging nuclide as a tracer in chemical oceanography, hydrology, and actinide sourcing. Depending on the properties of a sample and its neutron irradiation history, 236 U/ 238 U ratios from different sources vary significantly. Therefore, this ratio can be treated as an important fingerprint for radioactive source identification, and in particular, affords a definitive means of discriminating between naturally occurring U and specific types of anthropogenic U. The development of mass spectrometric techniques makes it possible to determine ultra-trace levels of 236 U in environmental samples. In this paper, we review the current status of mass spectrometric approaches for determination of 236 U in environmental samples. Various sample preparation methods are summarized and compared. The mass spectrometric techniques emphasized herein are thermal ionization mass spectrometry (TIMS), inductively coupled plasma mass spectrometry (ICP-MS) and accelerator mass spectrometry (AMS). The strategies or principles used by each technique for the analysis of 236 U are described. The performances of these techniques in terms of abundance sensitivity and detection limit are discussed in detail. To date, AMS exhibits the best capability for ultra-trace determinations of 236 U. The levels and behaviors of 236 U in various environmental media are summarized and discussed as well. Results suggest that 236 U has an important, emerging role as a tracer for geochemical studies. Copyright © 2017 Elsevier B.V. All rights reserved.

Analysis of sampling techniques for imbalanced data: An n = 648 ADNI study.

PubMed

Dubey, Rashmi; Zhou, Jiayu; Wang, Yalin; Thompson, Paul M; Ye, Jieping

2014-02-15

Many neuroimaging applications deal with imbalanced imaging data. For example, in Alzheimer's Disease Neuroimaging Initiative (ADNI) dataset, the mild cognitive impairment (MCI) cases eligible for the study are nearly two times the Alzheimer's disease (AD) patients for structural magnetic resonance imaging (MRI) modality and six times the control cases for proteomics modality. Constructing an accurate classifier from imbalanced data is a challenging task. Traditional classifiers that aim to maximize the overall prediction accuracy tend to classify all data into the majority class. In this paper, we study an ensemble system of feature selection and data sampling for the class imbalance problem. We systematically analyze various sampling techniques by examining the efficacy of different rates and types of undersampling, oversampling, and a combination of over and undersampling approaches. We thoroughly examine six widely used feature selection algorithms to identify significant biomarkers and thereby reduce the complexity of the data. The efficacy of the ensemble techniques is evaluated using two different classifiers including Random Forest and Support Vector Machines based on classification accuracy, area under the receiver operating characteristic curve (AUC), sensitivity, and specificity measures. Our extensive experimental results show that for various problem settings in ADNI, (1) a balanced training set obtained with K-Medoids technique based undersampling gives the best overall performance among different data sampling techniques and no sampling approach; and (2) sparse logistic regression with stability selection achieves competitive performance among various feature selection algorithms. Comprehensive experiments with various settings show that our proposed ensemble model of multiple undersampled datasets yields stable and promising results. © 2013 Elsevier Inc. All rights reserved.

FT-Raman and NIR spectroscopy data fusion strategy for multivariate qualitative analysis of food fraud.

PubMed

Márquez, Cristina; López, M Isabel; Ruisánchez, Itziar; Callao, M Pilar

2016-12-01

Two data fusion strategies (high- and mid-level) combined with a multivariate classification approach (Soft Independent Modelling of Class Analogy, SIMCA) have been applied to take advantage of the synergistic effect of the information obtained from two spectroscopic techniques: FT-Raman and NIR. Mid-level data fusion consists of merging some of the previous selected variables from the spectra obtained from each spectroscopic technique and then applying the classification technique. High-level data fusion combines the SIMCA classification results obtained individually from each spectroscopic technique. Of the possible ways to make the necessary combinations, we decided to use fuzzy aggregation connective operators. As a case study, we considered the possible adulteration of hazelnut paste with almond. Using the two-class SIMCA approach, class 1 consisted of unadulterated hazelnut samples and class 2 of samples adulterated with almond. Models performance was also studied with samples adulterated with chickpea. The results show that data fusion is an effective strategy since the performance parameters are better than the individual ones: sensitivity and specificity values between 75% and 100% for the individual techniques and between 96-100% and 88-100% for the mid- and high-level data fusion strategies, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

Evaluation of telomere length in human cardiac tissues using cardiac quantitative FISH.

PubMed

Sharifi-Sanjani, Maryam; Meeker, Alan K; Mourkioti, Foteini

2017-09-01

Telomere length has been correlated with various diseases, including cardiovascular disease and cancer. The use of currently available telomere-length measurement techniques is often restricted by the requirement of a large amount of cells (Southern-based techniques) or the lack of information on individual cells or telomeres (PCR-based methods). Although several methods have been used to measure telomere length in tissues as a whole, the assessment of cell-type-specific telomere length provides valuable information on individual cell types. The development of fluorescence in situ hybridization (FISH) technologies enables the quantification of telomeres in individual chromosomes, but the use of these methods is dependent on the availability of isolated cells, which prevents their use with fixed archival samples. Here we describe an optimized quantitative FISH (Q-FISH) protocol for measuring telomere length that bypasses the previous limitations by avoiding contributions from undesired cell types. We have used this protocol on small paraffin-embedded cardiac-tissue samples. This protocol describes step-by-step procedures for tissue preparation, permeabilization, cardiac-tissue pretreatment and hybridization with a Cy3-labeled telomeric repeat complementing (CCCTAA) 3 peptide nucleic acid (PNA) probe coupled with cardiac-specific antibody staining. We also describe how to quantify telomere length by means of the fluorescence intensity and area of each telomere within individual nuclei. This protocol provides comparative cell-type-specific telomere-length measurements in relatively small human cardiac samples and offers an attractive technique to test hypotheses implicating telomere length in various cardiac pathologies. The current protocol (from tissue collection to image procurement) takes ∼28 h along with three overnight incubations. We anticipate that the protocol could be easily adapted for use on different tissue types.

Specific detection of viable Listeria monocytogenes in Spanish wastewater treatment plants by Fluorescent In Situ Hybridization and PCR.

PubMed

Moreno, Yolanda; Ballesteros, Lorena; García-Hernández, Jorge; Santiago, Paula; González, Ana; Ferrús, M Antonia

2011-10-01

Listeria monocytogenes detection in wastewater can be difficult because of the large amount of background microbiota and the presence of viable but non-culturable forms in this environment. The aim of this study was to evaluate a Fluorescent In Situ Hybridization (FISH) assay combined with Direct Viable Count (DVC) method for detecting viable L. monocytogenes in wastewater samples, as an alternative to conventional culture methods. 16S rRNA sequence data were used to design a specific oligonucleotide probe. In order to assess the suitability of the method, the assays were performed on naturally (n=87) and artificially (n=14) contaminated samples and results were compared to those obtained with the isolation of cells on selective media and with a PCR method. The detection limit of FISH and PCR assays was 10(4) cells/mL without enrichment and 10 cells/mL after enrichment. A total of 47 samples, including 3 samples from effluent sites, yielded FISH positive results for L. monocytogenes. Using DVC-FISH technique, the presence of viable L. monocytogenes cells was detected in 23 out of these 47 FISH positive wastewater samples. PCR and culture methods yielded 27 and 23 positive results, respectively. According to these results, FISH technique has the potential to be used as a sensitive method for the detection and enumeration of L. monocytogenes in environmental wastewater samples. Copyright © 2011 Elsevier Ltd. All rights reserved.

Signal detection theory and vestibular perception: III. Estimating unbiased fit parameters for psychometric functions.

PubMed

Chaudhuri, Shomesh E; Merfeld, Daniel M

2013-03-01

Psychophysics generally relies on estimating a subject's ability to perform a specific task as a function of an observed stimulus. For threshold studies, the fitted functions are called psychometric functions. While fitting psychometric functions to data acquired using adaptive sampling procedures (e.g., "staircase" procedures), investigators have encountered a bias in the spread ("slope" or "threshold") parameter that has been attributed to the serial dependency of the adaptive data. Using simulations, we confirm this bias for cumulative Gaussian parametric maximum likelihood fits on data collected via adaptive sampling procedures, and then present a bias-reduced maximum likelihood fit that substantially reduces the bias without reducing the precision of the spread parameter estimate and without reducing the accuracy or precision of the other fit parameters. As a separate topic, we explain how to implement this bias reduction technique using generalized linear model fits as well as other numeric maximum likelihood techniques such as the Nelder-Mead simplex. We then provide a comparison of the iterative bootstrap and observed information matrix techniques for estimating parameter fit variance from adaptive sampling procedure data sets. The iterative bootstrap technique is shown to be slightly more accurate; however, the observed information technique executes in a small fraction (0.005 %) of the time required by the iterative bootstrap technique, which is an advantage when a real-time estimate of parameter fit variance is required.

First International External Quality Assessment Study on Molecular and Serological Methods for Yellow Fever Diagnosis

PubMed Central

Domingo, Cristina; Escadafal, Camille; Rumer, Leonid; Méndez, Jairo A.; García, Paquita; Sall, Amadou A.; Teichmann, Anette; Donoso-Mantke, Oliver; Niedrig, Matthias

2012-01-01

Objective We describe an external quality assurance (EQA) study designed to assess the efficiency and accurateness of molecular and serological methods used by expert laboratories performing YF diagnosis. Study Design For molecular diagnosis evaluation, a panel was prepared of 14 human plasma samples containing specific RNA of different YFV strains (YFV-17D, YFV South American strain [Brazil], YFV IvoryC1999 strain), and specificity samples containing other flaviviruses and negative controls. For the serological panel, 13 human plasma samples with anti-YFV-specific antibodies against different strains of YFV (YFV-17D strain, YFV IvoryC1999 strain, and YFV Brazilian strain), as well as specificity and negative controls, were included. Results Thirty-six laboratories from Europe, the Americas, Middle East, and Africa participated in these EQA activities. Only 16% of the analyses reported met all evaluation criteria with optimal performance. Serial dilutions of YFV-17D showed that in general the methodologies reported provided a suitable sensitivity. Failures were mainly due to the inability to detect wild-type strains or the presence of false positives. Performance in the serological diagnosis varied, mainly depending on the methodology used. Anti-YFV IgM detection was not performed in 16% of the reports using IIF or ELISA techniques, although it is preferable for the diagnosis of YFV acute infections. A good sensitivity profile was achieved in general; however, in the detection of IgM antibodies a lack of sensitivity of anti-YFV antibodies against the vaccine strain 17D was observed, and of the anti-YFV IgG antibodies against a West African strain. Neutralization assays showed a very good performance; however, the unexpected presence of false positives underlined the need of improving the running protocols. Conclusion This EQA provides information on each laboratory's efficacy of RT-PCR and serological YFV diagnosis techniques. The results indicate the need for improving serological and molecular diagnosis techniques and provide a follow-up of the diagnostic profiles. PMID:22570700

First international external quality assessment study on molecular and serological methods for yellow fever diagnosis.

PubMed

Domingo, Cristina; Escadafal, Camille; Rumer, Leonid; Méndez, Jairo A; García, Paquita; Sall, Amadou A; Teichmann, Anette; Donoso-Mantke, Oliver; Niedrig, Matthias

2012-01-01

We describe an external quality assurance (EQA) study designed to assess the efficiency and accurateness of molecular and serological methods used by expert laboratories performing YF diagnosis. For molecular diagnosis evaluation, a panel was prepared of 14 human plasma samples containing specific RNA of different YFV strains (YFV-17D, YFV South American strain [Brazil], YFV IvoryC1999 strain), and specificity samples containing other flaviviruses and negative controls. For the serological panel, 13 human plasma samples with anti-YFV-specific antibodies against different strains of YFV (YFV-17D strain, YFV IvoryC1999 strain, and YFV Brazilian strain), as well as specificity and negative controls, were included. Thirty-six laboratories from Europe, the Americas, Middle East, and Africa participated in these EQA activities. Only 16% of the analyses reported met all evaluation criteria with optimal performance. Serial dilutions of YFV-17D showed that in general the methodologies reported provided a suitable sensitivity. Failures were mainly due to the inability to detect wild-type strains or the presence of false positives. Performance in the serological diagnosis varied, mainly depending on the methodology used. Anti-YFV IgM detection was not performed in 16% of the reports using IIF or ELISA techniques, although it is preferable for the diagnosis of YFV acute infections. A good sensitivity profile was achieved in general; however, in the detection of IgM antibodies a lack of sensitivity of anti-YFV antibodies against the vaccine strain 17D was observed, and of the anti-YFV IgG antibodies against a West African strain. Neutralization assays showed a very good performance; however, the unexpected presence of false positives underlined the need of improving the running protocols. This EQA provides information on each laboratory's efficacy of RT-PCR and serological YFV diagnosis techniques. The results indicate the need for improving serological and molecular diagnosis techniques and provide a follow-up of the diagnostic profiles.

Design and implementation of coded aperture coherent scatter spectral imaging of cancerous and healthy breast tissue samples

PubMed Central

Lakshmanan, Manu N.; Greenberg, Joel A.; Samei, Ehsan; Kapadia, Anuj J.

2016-01-01

Abstract. A scatter imaging technique for the differentiation of cancerous and healthy breast tissue in a heterogeneous sample is introduced in this work. Such a technique has potential utility in intraoperative margin assessment during lumpectomy procedures. In this work, we investigate the feasibility of the imaging method for tumor classification using Monte Carlo simulations and physical experiments. The coded aperture coherent scatter spectral imaging technique was used to reconstruct three-dimensional (3-D) images of breast tissue samples acquired through a single-position snapshot acquisition, without rotation as is required in coherent scatter computed tomography. We perform a quantitative assessment of the accuracy of the cancerous voxel classification using Monte Carlo simulations of the imaging system; describe our experimental implementation of coded aperture scatter imaging; show the reconstructed images of the breast tissue samples; and present segmentations of the 3-D images in order to identify the cancerous and healthy tissue in the samples. From the Monte Carlo simulations, we find that coded aperture scatter imaging is able to reconstruct images of the samples and identify the distribution of cancerous and healthy tissues (i.e., fibroglandular, adipose, or a mix of the two) inside them with a cancerous voxel identification sensitivity, specificity, and accuracy of 92.4%, 91.9%, and 92.0%, respectively. From the experimental results, we find that the technique is able to identify cancerous and healthy tissue samples and reconstruct differential coherent scatter cross sections that are highly correlated with those measured by other groups using x-ray diffraction. Coded aperture scatter imaging has the potential to provide scatter images that automatically differentiate cancerous and healthy tissue inside samples within a time on the order of a minute per slice. PMID:26962543

Design and implementation of coded aperture coherent scatter spectral imaging of cancerous and healthy breast tissue samples.

PubMed

Lakshmanan, Manu N; Greenberg, Joel A; Samei, Ehsan; Kapadia, Anuj J

2016-01-01

A scatter imaging technique for the differentiation of cancerous and healthy breast tissue in a heterogeneous sample is introduced in this work. Such a technique has potential utility in intraoperative margin assessment during lumpectomy procedures. In this work, we investigate the feasibility of the imaging method for tumor classification using Monte Carlo simulations and physical experiments. The coded aperture coherent scatter spectral imaging technique was used to reconstruct three-dimensional (3-D) images of breast tissue samples acquired through a single-position snapshot acquisition, without rotation as is required in coherent scatter computed tomography. We perform a quantitative assessment of the accuracy of the cancerous voxel classification using Monte Carlo simulations of the imaging system; describe our experimental implementation of coded aperture scatter imaging; show the reconstructed images of the breast tissue samples; and present segmentations of the 3-D images in order to identify the cancerous and healthy tissue in the samples. From the Monte Carlo simulations, we find that coded aperture scatter imaging is able to reconstruct images of the samples and identify the distribution of cancerous and healthy tissues (i.e., fibroglandular, adipose, or a mix of the two) inside them with a cancerous voxel identification sensitivity, specificity, and accuracy of 92.4%, 91.9%, and 92.0%, respectively. From the experimental results, we find that the technique is able to identify cancerous and healthy tissue samples and reconstruct differential coherent scatter cross sections that are highly correlated with those measured by other groups using x-ray diffraction. Coded aperture scatter imaging has the potential to provide scatter images that automatically differentiate cancerous and healthy tissue inside samples within a time on the order of a minute per slice.

Current trends and challenges in sample preparation for metallic nanoparticles analysis in daily products and environmental samples: A review

NASA Astrophysics Data System (ADS)

De la Calle, Inmaculada; Menta, Mathieu; Séby, Fabienne

2016-11-01

Due to the increasing use of nanoparticles (NPs) in consumer products, it becomes necessary to develop different strategies for their detection, identification, characterization and quantification in a wide variety of samples. Since the analysis of NPs in consumer products and environmental samples is particularly troublesome, a detailed description of challenges and limitations is given here. This review mainly focuses on sample preparation procedures applied for the mostly used techniques for metallic and metal oxide NPs characterization in consumer products and most outstanding publications of biological and environmental samples (from 2006 to 2015). We summarize the procedures applied for total metal content, extraction/separation and/or preconcentration of NPs from the matrix, separation of metallic NPs from their ions or from larger particles and NPs' size fractionation. Sample preparation procedures specifically for microscopy are also described. Selected applications in cosmetics, food, other consumer products, biological tissues and environmental samples are presented. Advantages and inconveniences of those procedures are considered. Moreover, selected simplified schemes for NPs sample preparation, as well as usual techniques applied are included. Finally, promising directions for further investigations are discussed.

Accelerator mass spectrometry of small biological samples.

PubMed

Salehpour, Mehran; Forsgard, Niklas; Possnert, Göran

2008-12-01

Accelerator mass spectrometry (AMS) is an ultra-sensitive technique for isotopic ratio measurements. In the biomedical field, AMS can be used to measure femtomolar concentrations of labeled drugs in body fluids, with direct applications in early drug development such as Microdosing. Likewise, the regenerative properties of cells which are of fundamental significance in stem-cell research can be determined with an accuracy of a few years by AMS analysis of human DNA. However, AMS nominally requires about 1 mg of carbon per sample which is not always available when dealing with specific body substances such as localized, organ-specific DNA samples. Consequently, it is of analytical interest to develop methods for the routine analysis of small samples in the range of a few tens of microg. We have used a 5 MV Pelletron tandem accelerator to study small biological samples using AMS. Different methods are presented and compared. A (12)C-carrier sample preparation method is described which is potentially more sensitive and less susceptible to contamination than the standard procedures.

Preanalytical considerations in detection of colorectal cancer in blood serum using Raman molecular imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Treado, Patrick J.; Stewart, Shona D.; Smith, Aaron; Kirschner, Heather; Post, Christopher; Overholt, Bergein F.

2016-03-01

Colorectal cancer (CRC) is the third most common cancer in men and women in the United States. Raman Molecular Imaging (RMI) is an effective technique to evaluate human tissue, cells and bodily fluids, including blood serum for disease diagnosis. ChemImage Corporation, in collaboration with clinicians, has been engaged in development of an in vitro diagnostic Raman assay focused on CRC detection. The Raman Assay for Colorectal Cancer (RACC) exploits the high specificity of Raman imaging to distinguish diseased from normal dried blood serum droplets without additional reagents. Pilot Study results from testing of hundreds of biobank patient samples have demonstrated that RACC detects CRC with high sensitivity and specificity. However, expanded clinical trials, which are ongoing, are revealing a host of important preanalytical considerations associated with sample collection, sample storage and stability, sample shipping, sample preparation and sample interferents, which impact detection performance. Results from recent clinical studies will be presented.

Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping.

PubMed

Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A; Debelius, Justine; Morton, James T; Hyde, Embriette; Robbins-Pianka, Adam; Knight, Rob

2017-01-01

The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria , grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., "bloomed") at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called "Deblur" to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has been shown not to obscure physiologically relevant microbiome differences between comparison groups, there are still changes in specific bacterial taxa, notably, in members of the class Gammaproteobacteria , that can make microbiome profiles difficult to interpret. Here we identify the most problematic taxa and show that removing sequences from just a few fast-growing taxa is sufficient to correct microbiome profiles.

Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping

PubMed Central

Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A.; Debelius, Justine; Morton, James T.; Hyde, Embriette; Robbins-Pianka, Adam

2017-01-01

ABSTRACT The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria, grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., “bloomed”) at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called “Deblur” to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has been shown not to obscure physiologically relevant microbiome differences between comparison groups, there are still changes in specific bacterial taxa, notably, in members of the class Gammaproteobacteria, that can make microbiome profiles difficult to interpret. Here we identify the most problematic taxa and show that removing sequences from just a few fast-growing taxa is sufficient to correct microbiome profiles. PMID:28289733

A feasibility study for compressed sensing combined phase contrast MR angiography reconstruction

NASA Astrophysics Data System (ADS)

Lee, Dong-Hoon; Hong, Cheol-Pyo; Lee, Man-Woo; Han, Bong-Soo

2012-02-01

Phase contrast magnetic resonance angiography (PC MRA) is a technique for flow velocity measurement and vessels visualization, simultaneously. The PC MRA takes long scan time because each flow encoding gradients which are composed bipolar gradient type need to reconstruct the angiography image. Moreover, it takes more image acquisition time when we use the PC MRA at the low-tesla MRI system. In this study, we studied and evaluation of feasibility for CS MRI reconstruction combined PC MRA which data acquired by low-tesla MRI system. We used non-linear reconstruction algorithm which named Bregman iteration for CS image reconstruction and validate the usefulness of CS combined PC MRA reconstruction technique. The results of CS reconstructed PC MRA images provide similar level of image quality between fully sampled reconstruction data and sparse sampled reconstruction using CS technique. Although our results used half of sampling ratio and do not used specification hardware device or performance which are improving the temporal resolution of MR image acquisition such as parallel imaging reconstruction using phased array coil or non-cartesian trajectory, we think that CS combined PC MRA technique will be helpful to increase the temporal resolution and at low-tesla MRI system.

MUTAGENIC PROPERTIES OF RIVER WATERS FLOWING THROUGH BIG CITY AREAS IN NORTH AMERICA

EPA Science Inventory

The hanging technique using Blue rayon, which specifically adsorbs mutagens with multicyclic planer structures, has advantages over the conventional method of bringing-large volumes of water back to the laboratory for extraction. Therefore, the analysis of many samples from remot...

Effective Personnel Management. The 1980 Regional Meeting NACS Traveling Troupe Presentation.

ERIC Educational Resources Information Center

Arnett, Wayne; Williams, James L.

1981-01-01

Specific suggestions are given for more effective personnel management: careful screening of applicants by one person, a well-designed application form (a sample is provided), good interview techniques, job descriptions, promotions from within, employee manuals, employee orientation, and continuing interpersonal communication. (MSE)

Guidelines for collection and field analysis of water-quality samples from streams in Texas

USGS Publications Warehouse

Wells, F.C.; Gibbons, W.J.; Dorsey, M.E.

1990-01-01

Analyses for unstable constituents or properties are by necessity performed in the field. This manual addresses analytical techniques and quality assurance for: (1) Water temperature; (2) specific conductance; (3) pH; (4) alkalinity; (5) dissolved oxygen; and (6) bacteria.

Light chain typing of immunoglobulins in small samples of biological material

PubMed Central

Rádl, J.

1970-01-01

A method is described for the typing of the light chains of immunoglobulins in small samples of sera or external secretions and without their previous isolation. It consists of immunoelectrophoresis in agar plates which contain specific antisera against one of the light chain types. All immunoglobulins of this type are thus selected by precipitation in the central area during the electrophoretic phase. Immunoglobulins of the opposite light chain type diffuse through the agar and react with the class specific antisera from the troughs. This results in the precipitin lines as in conventional immunoelectrophoresis. This technique has proved most useful for typing heterogenous or homogeneous immunoglobulins in normal and low concentration. The antisera used for incorporation in the agar should fulfil special requirements. They should contain a high level of antibodies against common surface determinants of the immunoglobulin light chains. The further possibilities of this immunoselection technique for typing different protein mixtures is discussed. ImagesFIG. 1FIG. 2FIG. 3FIG. 4FIG. 5FIG. 6 PMID:4098592

Curve fitting air sample filter decay curves to estimate transuranic content.

PubMed

Hayes, Robert B; Chiou, Hung Cheng

2004-01-01

By testing industry standard techniques for radon progeny evaluation on air sample filters, a new technique is developed to evaluate transuranic activity on air filters by curve fitting the decay curves. The industry method modified here is simply the use of filter activity measurements at different times to estimate the air concentrations of radon progeny. The primary modification was to not look for specific radon progeny values but rather transuranic activity. By using a method that will provide reasonably conservative estimates of the transuranic activity present on a filter, some credit for the decay curve shape can then be taken. By carrying out rigorous statistical analysis of the curve fits to over 65 samples having no transuranic activity taken over a 10-mo period, an optimization of the fitting function and quality tests for this purpose was attained.

Diagnostic use of 3 techniques for identification of microsporidian spores among AIDS patients in Portugal.

PubMed

Matos, Olga; Lobo, Maria L; Gonçalves, Luzia; Antunes, Francisco

2002-01-01

The calcofluor stain (CF), the monoclonal antibody (MAb) 3B6 indirect immunofluorescence assay (IFA) and the modified trichrome blue stain (MT) were compared in terms of their reproducibility in a routine laboratory and in order to evaluate the percentage of cases of microsporidiosis in Portuguese HIV patients. A total of 166 faeces samples, 71 pulmonary specimens and 43 urine samples were studied using the 3 techniques. CF had a high sensitivity and a moderate specificity when applied to faeces samples. The sensitivity was lower with pulmonary specimens. The method is easy and quick to perform but readings take a long time to obtain. The MAb 3B6 IFA had a good to excellent sensitivity when applied to faeces and urine samples, but moderate sensitivity in pulmonary specimens. Readings were quick and easy to obtain, but the assay took longer to perform than the other 2 techniques. There was a greater correlation between the results obtained with the MT and MAb 3B6 IFA techniques than between those obtained with the MT and CF techniques. In conclusion, the MT performed better than the MAb 3B6 IFA and CF and continues to have an important place in a routine laboratory for the diagnosis of microsporidiosis. This work also confirms the existence of a relatively high proportion (30%) of cases of infection with Microsporidia, especially intestinal microsporidiosis, in HIV patients in Portugal.

Detection of circulating tumor cells from cryopreserved human sarcoma peripheral blood mononuclear cells.

PubMed

Li, Heming; Meng, Qing H; Noh, Hyangsoon; Batth, Izhar Singh; Somaiah, Neeta; Torres, Keila E; Xia, Xueqing; Wang, Ruoyu; Li, Shulin

2017-09-10

Circulating tumor cells (CTCs) enter the vasculature or lymphatic system after shedding from the primary tumor. CTCs may serve as "seed" cells for tumor metastasis. The utility of CTCs in clinical applications for sarcoma is not fully investigated, partly owing to the necessity for fresh blood samples and the lack of a CTC-specific antibody. To overcome these drawbacks, we developed a technique for sarcoma CTCs capture and detection using cryopreserved peripheral blood mononuclear cells (PBMCs) and our proprietary cell-surface vimentin (CSV) antibody 84-1, which is specific to tumor cells. This technique was validated by sarcoma cell spiking assay, matched CTCs comparison between fresh and cryopreserved PBMCs, and independent tumor markers in multiple types of sarcoma patient blood samples. The reproducibility was maximized when cryopreserved PBMCs were prepared from fresh blood samples within 2 h of the blood draw. In summary, as far as we are aware, ours is the first report to capture and detect CTCs from cryopreserved PBMCs. Further validation in other types of tumor may help boost the feasibility and utility of CTC-based diagnosis in a centralized laboratory. Copyright © 2017 Elsevier B.V. All rights reserved.

Validation of different techniques for the diagnosis of ventilator-associated pneumonia. Comparison with immediate postmortem pulmonary biopsy.

PubMed

Torres, A; el-Ebiary, M; Padró, L; Gonzalez, J; de la Bellacasa, J P; Ramirez, J; Xaubet, A; Ferrer, M; Rodriguez-Roisin, R

1994-02-01

To assess the accuracy of clinical parameters for the diagnosis of ventilator-associated (VA) pneumonia, as well as the diagnostic value of several invasive techniques, such as protected specimen brush (PSB), bronchoalveolar lavage (BAL), fiberoptic bronchial aspirates (FBAS), and percutaneous lung needle aspiration (PLNA), we compared the results of these techniques with the histopathology of immediate postmortem pulmonary biopsies, considered the "gold standard" reference test. We studied 30 mechanically ventilated patients (age 52 +/- 21 yr; mechanical ventilation period 9 +/- 7 days) who died in an intensive care unit. All patients received prior antibiotic treatment. The following procedures were performed immediately after death: bilateral PSB, BAL, FBAS, and PLNA, as well as bilateral minithoracotomies to obtain pulmonary biopsies as close as possible to the area sampled with the other techniques. According to the histopathology 18 patients had pneumonia and 12 did not. The presence of fever (sensitivity 55%, specificity 58%), purulent secretions (sensitivity 83%, specificity 33%), and chest radiograph infiltrates (sensitivity 78%, specificity 42%) could not differentiate in all instances presence from absence of pneumonia. Quantitative bacterial cultures of lung biopsies using 10(3) cfu/g as a cutoff point had low sensitivity (40%) and low specificity (45%) and could not differentiate the histologic absence or presence of pneumonia. Considering the histopathology of pulmonary biopsies as a gold standard, we found the following sensitivities for PSB, BAL, FBAS, and PLNA: 36, 50, 44, and 25%. The specificities were 50, 45, 48, and 79%, respectively. The sensitivities and specificities of different invasive techniques are much lower than those reported in clinical studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of the FilmArray blood culture identification panel for direct molecular diagnosis of infectious diseases from samples other than blood.

PubMed

Micó, Miquel; Navarro, Ferran; de Miniac, Daniela; González, Yésica; Brell, Albert; López, Cristina; Sánchez-Reus, Ferran; Mirelis, Beatriz; Coll, Pere

2015-12-01

Molecular-based techniques reduce the delay in diagnosing infectious diseases and therefore contribute to better patient outcomes. We assessed the FilmArray blood culture identification (BCID) panel (Biofire Diagnostics/bioMérieux) directly on clinical specimens other than blood: cerebrospinal, joint, pleural and ascitic fluids, bronchoscopy samples and abscesses. We compared the results from 88 samples obtained by culture-based techniques. The percentage of agreement between the two methods was 75 % with a Cohen κ value of 0.51. Global sensitivity and specificity using the FilmArray BCID panel were 71 and 97 %, respectively. Sensitivity was poorer in samples with a low bacterial load, such as ascitic and pleural fluids (25 %), whereas the sensitivity for abscess samples was high (89 %). These findings suggest that the FilmArray BCID panel could be useful to perform microbiological diagnosis directly from samples other than positive blood cultures, as it offers acceptable sensitivity and moderate agreement with conventional microbiological methods. Nevertheless, cost-benefit studies should be performed before introducing this method into algorithms for microbiological diagnostics.

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

PubMed

Ötles, Semih; Kartal, Canan

2016-01-01

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

Artist Material BRDF Database for Computer Graphics Rendering

NASA Astrophysics Data System (ADS)

Ashbaugh, Justin C.

The primary goal of this thesis was to create a physical library of artist material samples. This collection provides necessary data for the development of a gonio-imaging system for use in museums to more accurately document their collections. A sample set was produced consisting of 25 panels and containing nearly 600 unique samples. Selected materials are representative of those commonly used by artists both past and present. These take into account the variability in visual appearance resulting from the materials and application techniques used. Five attributes of variability were identified including medium, color, substrate, application technique and overcoat. Combinations of these attributes were selected based on those commonly observed in museum collections and suggested by surveying experts in the field. For each sample material, image data is collected and used to measure an average bi-directional reflectance distribution function (BRDF). The results are available as a public-domain image and optical database of artist materials at art-si.org. Additionally, the database includes specifications for each sample along with other information useful for computer graphics rendering such as the rectified sample images and normal maps.

Comparison of techniques for detecting antigens of Giardia lamblia and Cryptosporidium parvum in faeces.

PubMed Central

Tee, G H; Moody, A H; Cooke, A H; Chiodini, P L

1993-01-01

AIM--To compare the use of commercial monoclonal antibody test systems--the Giardia CEL IF test and the Crypto CEL IF test--for the detection of Giardia lamblia and Cryptosporidium parvum antigens in faeces with conventional techniques. METHODS--Sensitivity and specificity were evaluated using preparations of cysts of G lamblia and purified oocysts of C parvum. Evaluation of 59 random faecal samples passing through the Department of Clinical Parasitology, Hospital for Tropical Diseases, London, was carried out for both organisms. RESULTS--The fluorescence staining techniques proved more sensitive than other tests routinely used for diagnosis. PMID:8331181

A PCR method based on 18S rRNA gene for detection of malaria parasite in Balochistan.

PubMed

Shahwani, Zubeda; Aleem, Abdul; Ahmed, Nazeer; Mushtaq, Muhammad; Afridi, Sarwat

2016-12-01

To establish a polymerase chain reaction method based on 18S ribosomal ribonucleic acid gene for the detection of plasmodium deoxyribonucleic acid in patients suffering from malaria symptoms. This cross-sectional study was conducted from September 2013 to October 2014 in district Quetta of Pakistan's Balochistan province. Blood samples were collected from patients suffering from general symptoms of malaria. A polymerase chain reaction-based technique was applied for the diagnosis of malaria and detection of responsible species in the patients who were suspected to carry the parasite. Performance of this polymerase chain reaction method was compared against the microscopy results. Parasite number was also calculated for microscopy positive samples.All samples after the genomic deoxyribonucleic acid isolation were subjected to polymerase chain reaction amplification and agarose gel electrophoresis. Of the 200 samples, 114(57%) were confirmed as positive and 86(43%) as negative for malaria by microscopy. Polymerase chain reaction identified 124(62%) samples as positive and 76(38%) as negative for malaria. The comparative analysis of both diagnostic methods confirmed 109(54.5%) samples as positive by both techniques. Besides, 5(6.58%) samples were identified as false positive and 15(12.1%) samples as false negative by polymerase chain reaction. Sensitivity, specificity and positive predictive values for polymerase chain reaction in comparison to microscopy were 87.98%, 93.42% and 96%, respectively. Polymerase chain reaction-based methods in malaria diagnosis and species identification were found to be more effective than other techniques.

[Detection of Staphylococcus aureus resistant to methicillin (MRSA) by molecular biology (Cepheid GeneXpert IL, GeneOhm BD, Roche LightCycler, Hyplex Evigene I2A) versus screening by culture: Economic and practical strategy for the laboratory].

PubMed

Laudat, P; Demondion, E; Jouannet, C; Charron, J; Chillou, C; Salaun, V; Mankikian, B

2012-06-01

Patients admitted in cardiac surgery and cardiac ICU at the Clinic Saint-Gatien (Tours) are screened for MRSA at the entrance by nasal swab and culture on blood agar and selective chromogenic medium made by addition of cefoxitin: BBL CHROMagar MRSA-II BD (result obtained at Day +1). We wanted to assess the molecular biology techniques available to obtain a result at day 0 for the majority of patients and to define an economic and practical strategy for the laboratory. We studied four molecular biology techniques: Cepheid GeneXpert (Cepheid) GeneOhm (BD), LightCycler (Roche) and Hyplex (I2A). Upon reception, nasal swabs were treated by culture, considered as reference, and one of the techniques of molecular biology, according to the manufacturer's notice. We conducted four studies between April 2008 and February 2009 to obtain a significant sample for each of them. By screening we mean a method that allows us to exclude MRSA carriage for patients waiting for surgery, and not to change patient management: for example, lack of isolation measures specific to entrance, no modification of antibiotic prophylaxis during surgery and no isolation measures in the immediate postoperative period. The criteria we considered for this evaluation were: (1) technician time: time to perform one or a series of sample(s) n=10 or more (about 2h for all techniques except GeneXpert 75min), level of skilled competences (no specific training for GeneXpert); (2) results: turnaround time (all molecular biology techniques), ease of reading and results interpretations (no specialized training required for GeneXpert), failure or not (12% of failure of internal controls for GeneOhm); (3) economic: cost for one or a series of sample(s) (n=10 or more), if we considered X as the reference culture cost (10 X Hyplex and LightCycler, 20 X and 40 X for GeneXpert GeneOhm); (4) NPV: 100% for GeneXpert and LightCycler. At same sensitivity, no technique, including culture, can solve alone our problem, which is: (1) get results at day 0 for batch of samples (n<10): all molecular biology techniques; (2) beyond 10 samples: LightCycler (Roche) automated or Hyplex (I2A) manual; (3) when the result at day 1 is sufficient, the use of chromogenic agar with a reading of less than 18h as BBL CHROMagar MRSA II (BD) remains the most economical; (4) to be sure that a patient admitted at Day 0, even at night's emergency, is not carrier of MRSA: only Cepheid GeneXpert technology (IL). Furthermore, Cepheid GeneXpert (IL) allows performing several tests in parallel. The rapidity of this system can help control the transmission and make better use of antibiotics. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

COMPENDIUM OF METHODS FOR THE DETERMINATION ...

EPA Pesticide Factsheets

This Second Edition of the Compendium has been prepared to provide regional, state and local environmental regulatory agencies with step-by-step sampling and analysis procedures for the determination of selected toxic organic pollutants in ambient air. It is designed to assist those persons responsible for sampling and analysis of toxic organic pollutants in complying with the requirements of Title III of the Clean Air Act. This revised Compendium presents a set of 17 methods in a standardized format with a variety of applicable sampling methods, as well as several analytical techniques, for specific classes of organic pollutants, as appropriate to the specific pollutant compound, its level, and potential interferences. Consequently, this treatment allows the user flexibility in selecting alternatives to complement his or her background and laboratory capability. Information

A Novel Technique for the Connection of Ceramic and Titanium Implant Components Using Glass Solder Bonding

PubMed Central

Mick, Enrico; Tinschert, Joachim; Mitrovic, Aurica; Bader, Rainer

2015-01-01

Both titanium and ceramic materials provide specific advantages in dental implant technology. However, some problems, like hypersensitivity reactions, corrosion and mechanical failure, have been reported. Therefore, the combining of both materials to take advantage of their pros, while eliminating their respective cons, would be desirable. Hence, we introduced a new technique to bond titanium and ceramic materials by means of a silica-based glass ceramic solder. Cylindrical compound samples (Ø10 mm × 56 mm) made of alumina toughened zirconia (ATZ), as well as titanium grade 5, were bonded by glass solder on their end faces. As a control, a two-component adhesive glue was utilized. The samples were investigated without further treatment, after 30 and 90 days of storage in distilled water at room temperature, and after aging. All samples were subjected to quasi-static four-point-bending tests. We found that the glass solder bonding provided significantly higher bending strength than adhesive glue bonding. In contrast to the glued samples, the bending strength of the soldered samples remained unaltered by the storage and aging treatments. Scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analyses confirmed the presence of a stable solder-ceramic interface. Therefore, the glass solder technique represents a promising method for optimizing dental and orthopedic implant bondings. PMID:28793440

Fluorescence-based Western blotting for quantitation of protein biomarkers in clinical samples.

PubMed

Zellner, Maria; Babeluk, Rita; Diestinger, Michael; Pirchegger, Petra; Skeledzic, Senada; Oehler, Rudolf

2008-09-01

Since most high throughput techniques used in biomarker discovery are very time and cost intensive, highly specific and quantitative analytical alternative application methods are needed for the routine analysis. Conventional Western blotting allows detection of specific proteins to the level of single isotypes while its quantitative accuracy is rather limited. We report a novel and improved quantitative Western blotting method. The use of fluorescently labelled secondary antibodies strongly extends the dynamic range of the quantitation and improves the correlation with the protein amount (r=0.997). By an additional fluorescent staining of all proteins immediately after their transfer to the blot membrane, it is possible to visualise simultaneously the antibody binding and the total protein profile. This allows for an accurate correction for protein load. Applying this normalisation it could be demonstrated that fluorescence-based Western blotting is able to reproduce a quantitative analysis of two specific proteins in blood platelet samples from 44 subjects with different diseases as initially conducted by 2D-DIGE. These results show that the proposed fluorescence-based Western blotting is an adequate application technique for biomarker quantitation and suggest possibilities of employment that go far beyond.

Detection of Helicobacter pylori in drinking water treatment plants in Bogotá, Colombia, using cultural and molecular techniques.

PubMed

Vesga, Fidson-Juarismy; Moreno, Yolanda; Ferrús, María Antonia; Campos, Claudia; Trespalacios, Alba Alicia

2018-05-01

Helicobacter pylori is one of the most common causes of chronic bacterial infection in humans, and a predisposing factor for peptic ulcer and gastric cancer. The infection has been consistently associated with lack of access to clean water and proper sanitation. H. pylori has been detected in surface water, wastewater and drinking water. However, its ability to survive in an infectious state in the environment is hindered because it rapidly loses its cultivability. The aim of this study was to determine the presence of cultivable and therefore viable H. pylori in influent and effluent water from drinking water treatment plants (DWTP). A total of 310 influent and effluent water samples were collected from three drinking water treatment plants located at Bogotá city, Colombia. Specific detection of H. pylori was achieved by culture, qPCR and FISH techniques. Fifty-six positive H. pylori cultures were obtained from the water samples. Characteristic colonies were covered by the growth of a large number of other bacteria present in the water samples, making isolation difficult to perform. Thus, the mixed cultures were submitted to Fluorescent in situ Hybridization (FISH) and qPCR analysis, followed by sequencing of the amplicons for confirmation. By qPCR, 77 water samples, both from the influent and the effluent, were positive for the presence of H. pylori. The results of our study demonstrate that viable H. pylori cells were present in both, influent and effluent water samples obtained from drinking water treatment plants in Bogotá and provide further evidence that contaminated water may act as a transmission vehicle for H. pylori. Moreover, FISH and qPCR methods result rapid and specific techniques to identify H. pylori from complex environmental samples such as influent water. Copyright © 2018 Elsevier GmbH. All rights reserved.

Wetlands Research Program. Evaluation of Methods for Sampling Vegetation and Delineating Wetlands Transition Zones in Southern Louisiana, January 1979-May 1981.

DTIC Science & Technology

1983-10-01

sites should be collected and correlated to vegetational response through ordination or factor analysis techniques. 3. If site-specific methods are...specified number of samples is collected . If the 40 ."l ’ - - - .. -’-. - : -. .~ -, .4 .- . -.- , - ...... rb. . o* . -.% o - .- * .-.- v- . J...only, although the data collected were sufficient for determination of such parameters as density, cover, frequency, basal area, and importance value

New approaches to wipe sampling methods for antineoplastic and other hazardous drugs in healthcare settings.

PubMed

Connor, Thomas H; Smith, Jerome P

2016-09-01

At the present time, the method of choice to determine surface contamination of the workplace with antineoplastic and other hazardous drugs is surface wipe sampling and subsequent sample analysis with a variety of analytical techniques. The purpose of this article is to review current methodology for determining the level of surface contamination with hazardous drugs in healthcare settings and to discuss recent advances in this area. In addition it will provide some guidance for conducting surface wipe sampling and sample analysis for these drugs in healthcare settings. Published studies on the use of wipe sampling to measure hazardous drugs on surfaces in healthcare settings drugs were reviewed. These studies include the use of well-documented chromatographic techniques for sample analysis in addition to newly evolving technology that provides rapid analysis of specific antineoplastic. Methodology for the analysis of surface wipe samples for hazardous drugs are reviewed, including the purposes, technical factors, sampling strategy, materials required, and limitations. The use of lateral flow immunoassay (LFIA) and fluorescence covalent microbead immunosorbent assay (FCMIA) for surface wipe sample evaluation is also discussed. Current recommendations are that all healthc a re settings where antineoplastic and other hazardous drugs are handled include surface wipe sampling as part of a comprehensive hazardous drug-safe handling program. Surface wipe sampling may be used as a method to characterize potential occupational dermal exposure risk and to evaluate the effectiveness of implemented controls and the overall safety program. New technology, although currently limited in scope, may make wipe sampling for hazardous drugs more routine, less costly, and provide a shorter response time than classical analytical techniques now in use.

Autoantibodies against Leydig cells in patients after spermatic cord torsion.

PubMed Central

Zanchetta, R; Mastrogiacomo, I; Graziotti, P; Foresta, C; Betterle, C

1984-01-01

This study is aimed at searching for the presence of circulating antibodies against frozen sections of human testis, ovary and trophoblast in patients that had spermatic cord torsion. Sixty-eight sera samples were studied. Nine patients (13.2%) were positive for organ specific anti-testis autoantibodies. Six patients were positive for antibodies against Leydig cells: five were positive only with the indirect immunofluorescence technique of complement fixing (ITT/CF), the sixth patient was positive only with the indirect immunofluorescence technique (ITT). The other three patients were positive for antibodies against germ line cells: two patients were positive with both techniques, the third was positive only with indirect immunofluorescence technique. Eight of these patients were negative for antibodies against adrenal cortex while only one case was positive with indirect immunofluorescence technique both on adrenal cortex and Leydig cells. Human lyophilized testis absorbed the reactive antibodies against Leydig cells and germ line cells, while adrenal cortex and lyophilized testosterone were ineffective. This study shows the identification of a specific antibody against Leydig cells and germ line cells in patients after spermatic cord torsion. PMID:6362937

Detection and Serotyping of Dengue Virus in Serum Samples by Multiplex Reverse Transcriptase PCR-Ligase Detection Reaction Assay▿ †

PubMed Central

Das, S.; Pingle, M. R.; Muñoz-Jordán, J.; Rundell, M. S.; Rondini, S.; Granger, K.; Chang, G.-J. J.; Kelly, E.; Spier, E. G.; Larone, D.; Spitzer, E.; Barany, F.; Golightly, L. M.

2008-01-01

The detection and successful typing of dengue virus (DENV) from patients with suspected dengue fever is important both for the diagnosis of the disease and for the implementation of epidemiologic control measures. A technique for the multiplex detection and typing of DENV serotypes 1 to 4 (DENV-1 to DENV-4) from clinical samples by PCR-ligase detection reaction (LDR) has been developed. A serotype-specific PCR amplifies the regions of genes C and E simultaneously. The two amplicons are targeted in a multiplex LDR, and the resultant fluorescently labeled ligation products are detected on a universal array. The assay was optimized using 38 DENV strains and was evaluated with 350 archived acute-phase serum samples. The sensitivity of the assay was 98.7%, and its specificity was 98.4%, relative to the results of real-time PCR. The detection threshold was 0.017 PFU for DENV-1, 0.004 PFU for DENV-2, 0.8 PFU for DENV-3, and 0.7 PFU for DENV-4. The assay is specific; it does not cross-react with the other flaviviruses tested (West Nile virus, St. Louis encephalitis virus, Japanese encephalitis virus, Kunjin virus, Murray Valley virus, Powassan virus, and yellow fever virus). All but 1 of 26 genotypic variants of DENV serotypes in a global DENV panel from different geographic regions were successfully identified. The PCR-LDR assay is a rapid, sensitive, specific, and high-throughput technique for the simultaneous detection of all four serotypes of DENV. PMID:18685000

High-pressure sapphire cell for phase equilibria measurements of CO2/organic/water systems.

PubMed

Pollet, Pamela; Ethier, Amy L; Senter, James C; Eckert, Charles A; Liotta, Charles L

2014-01-24

The high pressure sapphire cell apparatus was constructed to visually determine the composition of multiphase systems without physical sampling. Specifically, the sapphire cell enables visual data collection from multiple loadings to solve a set of material balances to precisely determine phase composition. Ternary phase diagrams can then be established to determine the proportion of each component in each phase at a given condition. In principle, any ternary system can be studied although ternary systems (gas-liquid-liquid) are the specific examples discussed herein. For instance, the ternary THF-Water-CO2 system was studied at 25 and 40 °C and is described herein. Of key importance, this technique does not require sampling. Circumventing the possible disturbance of the system equilibrium upon sampling, inherent measurement errors, and technical difficulties of physically sampling under pressure is a significant benefit of this technique. Perhaps as important, the sapphire cell also enables the direct visual observation of the phase behavior. In fact, as the CO2 pressure is increased, the homogeneous THF-Water solution phase splits at about 2 MPa. With this technique, it was possible to easily and clearly observe the cloud point and determine the composition of the newly formed phases as a function of pressure. The data acquired with the sapphire cell technique can be used for many applications. In our case, we measured swelling and composition for tunable solvents, like gas-expanded liquids, gas-expanded ionic liquids and Organic Aqueous Tunable Systems (OATS)(1-4). For the latest system, OATS, the high-pressure sapphire cell enabled the study of (1) phase behavior as a function of pressure and temperature, (2) composition of each phase (gas-liquid-liquid) as a function of pressure and temperature and (3) catalyst partitioning in the two liquid phases as a function of pressure and composition. Finally, the sapphire cell is an especially effective tool to gather accurate and reproducible measurements in a timely fashion.

[Amanitine determination as an example of peptide analysis in the biological samples with HPLC-MS technique].

PubMed

Janus, Tomasz; Jasionowicz, Ewa; Potocka-Banaś, Barbara; Borowiak, Krzysztof

Routine toxicological analysis is mostly focused on the identification of non-organic and organic, chemically different compounds, but generally with low mass, usually not greater than 500–600 Da. Peptide compounds with atomic mass higher than 900 Da are a specific analytical group. Several dozen of them are highly-toxic substances well known in toxicological practice, for example mushroom toxin and animal venoms. In the paper the authors present an example of alpha-amanitin to explain the analytical problems and different original solutions in identifying peptides in urine samples with the use of the universal LC MS/MS procedure. The analyzed material was urine samples collected from patients with potential mushroom intoxication, routinely diagnosed for amanitin determination. Ultra filtration with centrifuge filter tubes (limited mass cutoff 3 kDa) was used. Filtrate fluid was directly injected on the chromatographic column and analyzed with a mass detector (MS/MS). The separation of peptides as organic, amphoteric compounds from biological material with the use of the SPE technique is well known but requires dedicated, specific columns. The presented paper proved that with the fast and simple ultra filtration technique amanitin can be effectively isolated from urine, and the procedure offers satisfactory sensitivity of detection and eliminates the influence of the biological matrix on analytical results. Another problem which had to be solved was the non-characteristic fragmentation of peptides in the MS/MS procedure providing non-selective chromatograms. It is possible to use higher collision energies in the analytical procedure, which results in more characteristic mass spectres, although it offers lower sensitivity. The ultra filtration technique as a procedure of sample preparation is effective for the isolation of amanitin from the biological matrix. The monitoring of selected mass corresponding to transition with the loss of water molecule offers satisfactory sensitivity of determination.

Western blotting using chemiluminescent substrates.

PubMed

Alegria-Schaffer, Alice

2014-01-01

Western blotting is a powerful and commonly used tool to identify and quantify a specific protein in a complex mixture (Towbin et al., 1979). The technique enables indirect detection of protein samples immobilized on a nitrocellulose or polyvinylidene fluoride (PVDF) membrane. Copyright © 2014 Elsevier Inc. All rights reserved.

Occurrence, Monitoring and Treatment of Cyanobacterial Toxins in Drinking Water

EPA Science Inventory

In the summer of 2014 a number of drinking water treatment plants (DWTPs) on Lake Erie supplied water samples on a monthly basis for analysis. Chlorophyll-a measurements, LC/MS/MS and ELISA techniques specific to microcystins were employed to measure potential harmful algal bloom...

An Object-Based Requirements Modeling Method.

ERIC Educational Resources Information Center

Cordes, David W.; Carver, Doris L.

1992-01-01

Discusses system modeling and specification as it relates to object-based information systems development and software development. An automated system model based on the objects in the initial requirements document is described, the requirements document translator is explained, and a sample application of the technique is provided. (12…

The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) identification versus biochemical tests: a study with enterobacteria from a dairy cattle environment.

PubMed

Rodrigues, Naiara Miranda Bento; Bronzato, Greiciane França; Santiago, Gabrielli Stefaninni; Botelho, Larissa Alvarenga Batista; Moreira, Beatriz Meurer; Coelho, Irene da Silva; Souza, Miliane Moreira Soares de; Coelho, Shana de Mattos de Oliveira

Mastitis adversely affects milk production and in general cows do not regain their full production levels post recovery, leading to considerable economic losses. Moreover the percentage decrease in milk production depends on the specific pathogen that caused the infection and enterobacteria are responsible for this greater reduction. Phenotypic tests are among the currently available methods used worldwide to identify enterobacteria; however they tend to misdiagnose the species despite the multiple tests carried out. On the other hand The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) technique has been attracting attention for its precise identification of several microorganisms at species level. In the current study, 183 enterobacteria were detected in milk (n=47) and fecal samples (n=94) from cows, and samples from water (n=23) and milk lines (n=19). All these samples were collected from a farm in Rio de Janeiro with the specific purpose of presenting the MALDI-TOF MS technique as an efficient methodology to identify Enterobacteriaceae from bovine environments. The MALDI-TOF MS technique results matched the biochemical test results in 92.9% (170/183) of the enterobacteria species and the gyrB sequencing confirmed 100% of the proteomic technique results. The amino acid decarboxylation test made the most misidentifications and Enterobacter spp. was the most misidentified genus (76.9%, 10/13). These results aim to clarify the current biochemical errors in enterobacteria identification, considering isolates from a bovine environment, and show the importance for more careful readings of phenotypic tests which are often used in veterinary microbiology laboratories. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

The Effect of Different Mixing Methods on Working Time, Setting Time, Dimensional Changes and Film Thickness of Mineral Trioxide Aggregate and Calcium-Enriched Mixture.

PubMed

Shahi, Shahriar; Ghasemi, Negin; Rahimi, Saeed; Yavari, Hamidreza; Janani, Maryam; Mokhtari, Hadi; Bahari, Mahmood; Rabbani, Parastu

2015-01-01

The aim of the present study was to evaluate the effect of different mixing techniques (conventional, amalgamator and ultrasonic mixing) on the physical properties the working time (WT), setting time (ST), dimensional changes (DC) and film thickness (FT)] of calcium-enriched mixture (CEM) cement and mineral trioxide aggregate (MTA). The mentioned physical properties were determined using the ISO 6786:2001 specification. Six samples of each material were prepared for three mixing techniques (totally 36 samples). Data were analyzed using descriptive statistics, two-way ANOVA and Post Hoc Tukey's tests. The level of significance was defined at 0.05. Irrespective of mixing technique, there was no significant difference between the WT and FT of the tested materials. Except for the DC of MTA and the FT of the all materials, other properties were significantly affected with mixing techniques (P<0.05). The ultrasonic technique decreased the ST of MTA and CEM cement and increased the WT of CEM cement (P<0.05). The mixing technique of the materials had no significant effect on the dimensional changes of MTA and the film thickness of both materials.

Application of Structured Light System Technique for Authentication of Wooden Panel Paintings.

PubMed

Buchón-Moragues, Fernando; Bravo, José María; Ferri, Marcelino; Redondo, Javier; Sánchez-Pérez, Juan Vicente

2016-06-14

This paper presents a new application of photogrammetric techniques for protecting cultural heritage. The accuracy of the method and the fact that it can be used to carry out different tests without contact between the sample and the instruments can make this technique very useful for authenticating and cataloging artworks. The application focuses on the field of pictorial artworks, and wooden panel paintings in particular. In these works, the orography formed by the brushstrokes can be easily digitalized using a photogrammetric technique, called Structured Light System, with submillimeter accuracy. Thus, some of the physical characteristics of the brushstrokes, like minimum and maximum heights or slopes become a fingerprint of the painting. We explain in detail the general principles of the Structured Light System Technique and the specific characteristics of the commercial set-up used in this work. Some experiments are carried out on a sample painted by us to check the accuracy limits of the technique and to propose some tests that can help to stablish a methodology for authentication purposes. Finally, some preliminary results obtained on a real pictorial artwork are presented, providing geometrical information of its metric features as an example of the possibilities of this application.

Application of Structured Light System Technique for Authentication of Wooden Panel Paintings

PubMed Central

Buchón-Moragues, Fernando; Bravo, José María; Ferri, Marcelino; Redondo, Javier; Sánchez-Pérez, Juan Vicente

2016-01-01

This paper presents a new application of photogrammetric techniques for protecting cultural heritage. The accuracy of the method and the fact that it can be used to carry out different tests without contact between the sample and the instruments can make this technique very useful for authenticating and cataloging artworks. The application focuses on the field of pictorial artworks, and wooden panel paintings in particular. In these works, the orography formed by the brushstrokes can be easily digitalized using a photogrammetric technique, called Structured Light System, with submillimeter accuracy. Thus, some of the physical characteristics of the brushstrokes, like minimum and maximum heights or slopes become a fingerprint of the painting. We explain in detail the general principles of the Structured Light System Technique and the specific characteristics of the commercial set-up used in this work. Some experiments are carried out on a sample painted by us to check the accuracy limits of the technique and to propose some tests that can help to stablish a methodology for authentication purposes. Finally, some preliminary results obtained on a real pictorial artwork are presented, providing geometrical information of its metric features as an example of the possibilities of this application. PMID:27314353

Intracavity optogalvanic spectroscopy. An analytical technique for 14C analysis with subattomole sensitivity.

PubMed

Murnick, Daniel E; Dogru, Ozgur; Ilkmen, Erhan

2008-07-01

We show a new ultrasensitive laser-based analytical technique, intracavity optogalvanic spectroscopy, allowing extremely high sensitivity for detection of (14)C-labeled carbon dioxide. Capable of replacing large accelerator mass spectrometers, the technique quantifies attomoles of (14)C in submicrogram samples. Based on the specificity of narrow laser resonances coupled with the sensitivity provided by standing waves in an optical cavity and detection via impedance variations, limits of detection near 10(-15) (14)C/(12)C ratios are obtained. Using a 15-W (14)CO2 laser, a linear calibration with samples from 10(-15) to >1.5 x 10(-12) in (14)C/(12)C ratios, as determined by accelerator mass spectrometry, is demonstrated. Possible applications include microdosing studies in drug development, individualized subtherapeutic tests of drug metabolism, carbon dating and real time monitoring of atmospheric radiocarbon. The method can also be applied to detection of other trace entities.

Online immunoaffinity LC/MS/MS. A general method to increase sensitivity and specificity: How do you do it and what do you need?

PubMed

Dufield, Dawn R; Radabaugh, Melissa R

2012-02-01

There is an increased emphasis on hyphenated techniques such as immunoaffinity LC/MS/MS (IA-LC/MS/MS) or IA-LC/MRM. These techniques offer competitive advantages with respect to sensitivity and selectivity over traditional LC/MS and are complementary to ligand binding assays (LBA) or ELISA's. However, these techniques are not entirely straightforward and there are several tips and tricks to routine sample analysis. We describe here our methods and procedures for how to perform online IA-LC/MS/MS including a detailed protocol for the preparation of antibody (Ab) enrichment columns. We have included sample trapping and Ab methods. Furthermore, we highlight tips, tricks, minimal and optimal approaches. This technology has been shown to be viable for several applications, species and fluids from small molecules to proteins and biomarkers to PK assays. Copyright © 2011 Elsevier Inc. All rights reserved.

Evaluation and study of advanced optical contamination, deposition, measurement, and removal techniques. [including computer programs and ultraviolet reflection analysis

NASA Technical Reports Server (NTRS)

Linford, R. M. F.; Allen, T. H.; Dillow, C. F.

1975-01-01

A program is described to design, fabricate and install an experimental work chamber assembly (WCA) to provide a wide range of experimental capability. The WCA incorporates several techniques for studying the kinetics of contaminant films and their effect on optical surfaces. It incorporates the capability for depositing both optical and contaminant films on temperature-controlled samples, and for in-situ measurements of the vacuum ultraviolet reflectance. Ellipsometer optics are mounted on the chamber for film thickness determinations, and other features include access ports for radiation sources and instrumentation. Several supporting studies were conducted to define specific chamber requirements, to determine the sensitivity of the measurement techniques to be incorporated in the chamber, and to establish procedures for handling samples prior to their installation in the chamber. A bibliography and literature survey of contamination-related articles is included.

X-ray near-field speckle: implementation and critical analysis

PubMed Central

Lu, Xinhui; Mochrie, S. G. J.; Narayanan, S.; Sandy, A. R.; Sprung, M.

2011-01-01

The newly introduced coherence-based technique of X-ray near-field speckle (XNFS) has been implemented at 8-ID-I at the Advanced Photon Source. In the near-field regime of high-brilliance synchrotron X-rays scattered from a sample of interest, it turns out that, when the scattered radiation and the main beam both impinge upon an X-ray area detector, the measured intensity shows low-contrast speckles, resulting from interference between the incident and scattered beams. A micrometer-resolution XNFS detector with a high numerical aperture microscope objective has been built and its capability for studying static structures and dynamics at longer length scales than traditional far-field X-ray scattering techniques is demonstrated. Specifically, the dynamics of dilute silica and polystyrene colloidal samples are characterized. This study reveals certain limitations of the XNFS technique, especially in the characterization of static structures, which is discussed. PMID:21997906

Differentiation of Candida albicans, Candida glabrata, and Candida krusei by FT-IR and chemometrics by CHROMagar™ Candida.

PubMed

Wohlmeister, Denise; Vianna, Débora Renz Barreto; Helfer, Virginia Etges; Calil, Luciane Noal; Buffon, Andréia; Fuentefria, Alexandre Meneghello; Corbellini, Valeriano Antonio; Pilger, Diogo André

2017-10-01

Pathogenic Candida species are detected in clinical infections. CHROMagar™ is a phenotypical method used to identify Candida species, although it has limitations, which indicates the need for more sensitive and specific techniques. Infrared Spectroscopy (FT-IR) is an analytical vibrational technique used to identify patterns of metabolic fingerprint of biological matrixes, particularly whole microbial cell systems as Candida sp. in association of classificatory chemometrics algorithms. On the other hand, Soft Independent Modeling by Class Analogy (SIMCA) is one of the typical algorithms still little employed in microbiological classification. This study demonstrates the applicability of the FT-IR-technique by specular reflectance associated with SIMCA to discriminate Candida species isolated from vaginal discharges and grown on CHROMagar™. The differences in spectra of C. albicans, C. glabrata and C. krusei were suitable for use in the discrimination of these species, which was observed by PCA. Then, a SIMCA model was constructed with standard samples of three species and using the spectral region of 1792-1561cm -1 . All samples (n=48) were properly classified based on the chromogenic method using CHROMagar™ Candida. In total, 93.4% (n=45) of the samples were correctly and unambiguously classified (Class I). Two samples of C. albicans were classified correctly, though these could have been C. glabrata (Class II). Also, one C. glabrata sample could have been classified as C. krusei (Class II). Concerning these three samples, one triplicate of each was included in Class II and two in Class I. Therefore, FT-IR associated with SIMCA can be used to identify samples of C. albicans, C. glabrata, and C. krusei grown in CHROMagar™ Candida aiming to improve clinical applications of this technique. Copyright © 2017 Elsevier B.V. All rights reserved.

A study on Improvisation in a Musical performance using Multifractal Detrended Cross Correlation Analysis

NASA Astrophysics Data System (ADS)

Sanyal, Shankha; Banerjee, Archi; Patranabis, Anirban; Banerjee, Kaushik; Sengupta, Ranjan; Ghosh, Dipak

2016-11-01

MFDFA (the most rigorous technique to assess multifractality) was performed on four Hindustani music samples played on same 'raga' sung by the same performer. Each music sample was divided into six parts and 'multifractal spectral width' was determined for each part corresponding to the four samples. The results obtained reveal that different parts of all the four sound signals possess spectral width of widely varying values. This gives a cue of the so called 'musical improvisation' in all music samples, keeping in mind they belong to the bandish part of the same raga. Formal compositions in Hindustani raga are juxtaposed with the improvised portions, where an artist manoeuvers his/her own creativity to bring out a mood that is specific for that particular performance, which is known as 'improvisation'. Further, this observation hints at the association of different emotions even in the same bandish of the same raga performed by the same artist, this interesting observation cannot be revealed unless rigorous non-linear technique explores the nature of musical structure. In the second part, we applied MFDXA technique to explore more in-depth about 'improvisation' and association with emotion. This technique is applied to find the degree of cross-correlation (γx) between the different parts of the samples. Pronounced correlation has been observed in the middle parts of the all the four samples evident from higher values of γx ​whereas the other parts show weak correlation. This gets further support from the values of spectral width from different parts of the sample - width of those parts is significantly different from other parts. This observation is extremely new both in respect of musical structure of so called improvisation and associated emotion. The importance of this study in application area of cognitive music therapy is immense.

Cell-type-specific profiling of protein-DNA interactions without cell isolation using targeted DamID with next-generation sequencing.

PubMed

Marshall, Owen J; Southall, Tony D; Cheetham, Seth W; Brand, Andrea H

2016-09-01

This protocol is an extension to: Nat. Protoc. 2, 1467-1478 (2007); doi:10.1038/nprot.2007.148; published online 7 June 2007The ability to profile transcription and chromatin binding in a cell-type-specific manner is a powerful aid to understanding cell-fate specification and cellular function in multicellular organisms. We recently developed targeted DamID (TaDa) to enable genome-wide, cell-type-specific profiling of DNA- and chromatin-binding proteins in vivo without cell isolation. As a protocol extension, this article describes substantial modifications to an existing protocol, and it offers additional applications. TaDa builds upon DamID, a technique for detecting genome-wide DNA-binding profiles of proteins, by coupling it with the GAL4 system in Drosophila to enable both temporal and spatial resolution. TaDa ensures that Dam-fusion proteins are expressed at very low levels, thus avoiding toxicity and potential artifacts from overexpression. The modifications to the core DamID technique presented here also increase the speed of sample processing and throughput, and adapt the method to next-generation sequencing technology. TaDa is robust, reproducible and highly sensitive. Compared with other methods for cell-type-specific profiling, the technique requires no cell-sorting, cross-linking or antisera, and binding profiles can be generated from as few as 10,000 total induced cells. By profiling the genome-wide binding of RNA polymerase II (Pol II), TaDa can also identify transcribed genes in a cell-type-specific manner. Here we describe a detailed protocol for carrying out TaDa experiments and preparing the material for next-generation sequencing. Although we developed TaDa in Drosophila, it should be easily adapted to other organisms with an inducible expression system. Once transgenic animals are obtained, the entire experimental procedure-from collecting tissue samples to generating sequencing libraries-can be accomplished within 5 d.

A Sampling Based Approach to Spacecraft Autonomous Maneuvering with Safety Specifications

NASA Technical Reports Server (NTRS)

Starek, Joseph A.; Barbee, Brent W.; Pavone, Marco

2015-01-01

This paper presents a methods for safe spacecraft autonomous maneuvering that leverages robotic motion-planning techniques to spacecraft control. Specifically the scenario we consider is an in-plan rendezvous of a chaser spacecraft in proximity to a target spacecraft at the origin of the Clohessy Wiltshire Hill frame. The trajectory for the chaser spacecraft is generated in a receding horizon fashion by executing a sampling based robotic motion planning algorithm name Fast Marching Trees (FMT) which efficiently grows a tree of trajectories over a set of probabillistically drawn samples in the state space. To enforce safety the tree is only grown over actively safe samples for which there exists a one-burn collision avoidance maneuver that circularizes the spacecraft orbit along a collision-free coasting arc and that can be executed under potential thrusters failures. The overall approach establishes a provably correct framework for the systematic encoding of safety specifications into the spacecraft trajectory generations process and appears amenable to real time implementation on orbit. Simulation results are presented for a two-fault tolerant spacecraft during autonomous approach to a single client in Low Earth Orbit.

Digital PCR quantification of MGMT methylation refines prediction of clinical benefit from alkylating agents in glioblastoma and metastatic colorectal cancer.

PubMed

Barault, L; Amatu, A; Bleeker, F E; Moutinho, C; Falcomatà, C; Fiano, V; Cassingena, A; Siravegna, G; Milione, M; Cassoni, P; De Braud, F; Rudà, R; Soffietti, R; Venesio, T; Bardelli, A; Wesseling, P; de Witt Hamer, P; Pietrantonio, F; Siena, S; Esteller, M; Sartore-Bianchi, A; Di Nicolantonio, F

2015-09-01

O(6)-methyl-guanine-methyl-transferase (MGMT) silencing by promoter methylation may identify cancer patients responding to the alkylating agents dacarbazine or temozolomide. We evaluated the prognostic and predictive value of MGMT methylation testing both in tumor and cell-free circulating DNA (cfDNA) from plasma samples using an ultra-sensitive two-step digital PCR technique (methyl-BEAMing). Results were compared with two established techniques, methylation-specific PCR (MSP) and Bs-pyrosequencing. Thresholds for MGMT methylated status for each technique were established in a training set of 98 glioblastoma (GBM) patients. The prognostic and the predictive value of MGMT methylated status was validated in a second cohort of 66 GBM patients treated with temozolomide in which methyl-BEAMing displayed a better specificity than the other techniques. Cutoff values of MGMT methylation specific for metastatic colorectal cancer (mCRC) tissue samples were established in a cohort of 60 patients treated with dacarbazine. In mCRC, both quantitative assays methyl-BEAMing and Bs-pyrosequencing outperformed MSP, providing better prediction of treatment response and improvement in progression-free survival (PFS) (P < 0.001). Ability of methyl-BEAMing to identify responding patients was validated in a cohort of 23 mCRC patients treated with temozolomide and preselected for MGMT methylated status according to MSP. In mCRC patients treated with dacarbazine, exploratory analysis of cfDNA by methyl-BEAMing showed that MGMT methylation was associated with better response and improved median PFS (P = 0.008). Methyl-BEAMing showed high reproducibility, specificity and sensitivity and was applicable to formalin-fixed paraffin-embedded tissues and cfDNA. This study supports the quantitative assessment of MGMT methylation for clinical purposes since it could refine prediction of response to alkylating agents. © The Author 2015. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Auditory steady-state response in cochlear implant patients.

PubMed

Torres-Fortuny, Alejandro; Arnaiz-Marquez, Isabel; Hernández-Pérez, Heivet; Eimil-Suárez, Eduardo

2018-03-19

Auditory steady state responses to continuous amplitude modulated tones at rates between 70 and 110Hz, have been proposed as a feasible alternative to objective frequency specific audiometry in cochlear implant subjects. The aim of the present study is to obtain physiological thresholds by means of auditory steady-state response in cochlear implant patients (Clarion HiRes 90K), with acoustic stimulation, on free field conditions and to verify its biological origin. 11 subjects comprised the sample. Four amplitude modulated tones of 500, 1000, 2000 and 4000Hz were used as stimuli, using the multiple frequency technique. The recording of auditory steady-state response was also recorded at 0dB HL of intensity, non-specific stimulus and using a masking technique. The study enabled the electrophysiological thresholds to be obtained for each subject of the explored sample. There were no auditory steady-state responses at either 0dB or non-specific stimulus recordings. It was possible to obtain the masking thresholds. A difference was identified between behavioral and electrophysiological thresholds of -6±16, -2±13, 0±22 and -8±18dB at frequencies of 500, 1000, 2000 and 4000Hz respectively. The auditory steady state response seems to be a suitable technique to evaluate the hearing threshold in cochlear implant subjects. Copyright © 2018 Sociedad Española de Otorrinolaringología y Cirugía de Cabeza y Cuello. Publicado por Elsevier España, S.L.U. All rights reserved.

Preservation of capsular material of streptococcal cells by specific lectins determined by immunoelectron microscopy.

PubMed

Molinari, A; Orefici, G; Donelli, G; Von Hunolstein, C; Paradisi, S; Arancia, G

1988-09-01

We describe the use of lectins as specific stabilizing agents for the polysaccharide capsular components of two Gram-positive bacteria, Streptococcus agalactiae and Streptococcus bovis. Treatment of bacterial suspensions with wheatgerm agglutinin and concanavalin A allowed better morphological preservation as well as immunoelectron microscopic localization of a capsular component (lipoteichoic acid) by employing specific antibodies and the protein A-gold technique. Data obtained indicate that lectins are useful agents in preserving highly water-soluble capsular components during the electron microscopy procedures for both unembedded and embedded samples.

The application of supported liquid extraction in the analysis of benzodiazepines using surface enhanced Raman spectroscopy.

PubMed

Doctor, Erika L; McCord, Bruce

2015-11-01

Benzodiazepines are among the most frequently prescribed medicines for anxiety disorders and are present in many toxicological screens. These drugs are often administered in the commission of drug facilitated sexual assaults due their effects on the central nervous system. Due to the potency of the drugs, only small amounts are usually given to victims; therefore, the target detection limit for these compounds in biological samples has been set at 50 ng/mL. Currently the standard screening method for detection of this class of drug is the immunoassay; however, screening methods that are more sensitive and selective than immunoassays are needed to encompass the wide range of structural variants of this class of compounds. Surface enhanced Raman spectroscopy (SERS) can be highly sensitive and has been shown to permit analysis of various benzodiazepines with limits of detection as low as 6 ng/mL. This technique permits analytical results in less than 2 min when used on pure drug samples. For biological samples, a key issue for analysis by SERS is removal of exogenous salts and matrix components. In this paper we examine supported liquid extraction as a useful preparation technique for SERS detection. Supported liquid extraction has many of the benefits of liquid-liquid extraction along with the ability to be automated. This technique provides a fast and clean extraction for benzodiazepines from urine at a pH of 5.0, and does not produce large quantities of solvent waste. To validate this procedure we have determined figures of merit and examined simulated urine samples prepared with commonly appearing interferences. It was shown that at a pH 5.0 many drugs that are prevalent in urine samples can be removed, permitting a selective detection of the benzodiazepine of interest. This technique has been shown to provide rapid (less than 20 min), sensitive, and specific detection of benzodiazepines with limits of detection between 32 and 600 ng/mL and dynamic range of 32-25,000 ng/mL. It provides the forensic community with a sensitive and specific screening technique for the detection of benzodiazepines in drug facilitated assault cases. Copyright © 2015 Elsevier B.V. All rights reserved.

SVM classification of microaneurysms with imbalanced dataset based on borderline-SMOTE and data cleaning techniques

NASA Astrophysics Data System (ADS)

Wang, Qingjie; Xin, Jingmin; Wu, Jiayi; Zheng, Nanning

2017-03-01

Microaneurysms are the earliest clinic signs of diabetic retinopathy, and many algorithms were developed for the automatic classification of these specific pathology. However, the imbalanced class distribution of dataset usually causes the classification accuracy of true microaneurysms be low. Therefore, by combining the borderline synthetic minority over-sampling technique (BSMOTE) with the data cleaning techniques such as Tomek links and Wilson's edited nearest neighbor rule (ENN) to resample the imbalanced dataset, we propose two new support vector machine (SVM) classification algorithms for the microaneurysms. The proposed BSMOTE-Tomek and BSMOTE-ENN algorithms consist of: 1) the adaptive synthesis of the minority samples in the neighborhood of the borderline, and 2) the remove of redundant training samples for improving the efficiency of data utilization. Moreover, the modified SVM classifier with probabilistic outputs is used to divide the microaneurysm candidates into two groups: true microaneurysms and false microaneurysms. The experiments with a public microaneurysms database shows that the proposed algorithms have better classification performance including the receiver operating characteristic (ROC) curve and the free-response receiver operating characteristic (FROC) curve.

New method of measuring low values of dielectric loss in the near millimetre wavelength region using untuned cavities

NASA Astrophysics Data System (ADS)

Llewellyn-Jones, D. T.; Knight, R. J.; Moffat, P. H.; Gebbie, H. A.

1980-11-01

In the near millimeter-wavelength region, low values of dielectric loss in a material can be readily measured by inserting a sample into an untuned cavity resonator. The high-Q values of the cavities give the technique great sensitivity to low values of loss tangent and, in contrast to other techniques, place very few restrictions on the shape, size, and position of the sample. The technique is demonstrated by measurements at 156 GHz on several polymer materials whose low loss factors are of practical interest. It is shown that the loading of an untuned cavity by a solid sample of low loss is proportional to its absorption cross section, which is the product of its volume and its linear absorption coefficient in the trivial case of n = 1. In the usual case of n greater than 1, reflection at the boundaries will affect the measured cross section in a way that has been investigated experimentally for a number of shapes, both simple and complex, and theoretically for the specific cases of slabs and cubes.

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.

Molecular diagnostics for the detection of Bokeloh bat lyssavirus in a bat from Bavaria, Germany.

PubMed

Freuling, Conrad M; Abendroth, Björn; Beer, Martin; Fischer, Melina; Hanke, Dennis; Hoffmann, Bernd; Höper, Dirk; Just, Frank; Mettenleiter, Thomas C; Schatz, Juliane; Müller, Thomas

2013-11-06

A brain sample of a Natterer's bat tested positive for rabies with classical virological techniques. Molecular techniques confirmed the presence of Bokeloh bat lyssavirus (BBLV) in Germany for the second time. Sequence analysis revealed a close genetic relationship to the initial German BBLV case. Using a TaqMan RT-PCR specific for BBLV viral RNA was detected in various other organs albeit with differences in the relative viral load. Copyright © 2013 Elsevier B.V. All rights reserved.

Analysis of Ergot Alkaloids

PubMed Central

Crews, Colin

2015-01-01

The principles and application of established and newer methods for the quantitative and semi-quantitative determination of ergot alkaloids in food, feed, plant materials and animal tissues are reviewed. The techniques of sampling, extraction, clean-up, detection, quantification and validation are described. The major procedures for ergot alkaloid analysis comprise liquid chromatography with tandem mass spectrometry (LC-MS/MS) and liquid chromatography with fluorescence detection (LC-FLD). Other methods based on immunoassays are under development and variations of these and minor techniques are available for specific purposes. PMID:26046699

Chi-squared and C statistic minimization for low count per bin data

NASA Astrophysics Data System (ADS)

Nousek, John A.; Shue, David R.

1989-07-01

Results are presented from a computer simulation comparing two statistical fitting techniques on data samples with large and small counts per bin; the results are then related specifically to X-ray astronomy. The Marquardt and Powell minimization techniques are compared by using both to minimize the chi-squared statistic. In addition, Cash's C statistic is applied, with Powell's method, and it is shown that the C statistic produces better fits in the low-count regime than chi-squared.

Qualitative evaluation of maternal milk and commercial infant formulas via LIBS.

PubMed

Abdel-Salam, Z; Al Sharnoubi, J; Harith, M A

2013-10-15

This study focuses on the use of laser-induced breakdown spectroscopy (LIBS) for the evaluation of the nutrients in maternal milk and some commercially available infant formulas. The results of such evaluation are vital for adequate and healthy feeding for babies during lactation period. Laser-induced breakdown spectroscopy offers special advantages in comparison to the other conventional analytical techniques. Specifically, LIBS is a straightforward technique that can be used in situ to provide qualitative analytical information in few minutes for the samples under investigation without preparation processes. The samples studied in the current work were maternal milk samples collected during the first 3 months of lactation (not colostrum milk) and samples from six different types of commercially available infant formulas. The samples' elemental composition has been compared with respect to the relative abundance of the elements of nutrition importance, namely Mg, Ca, Na, and Fe using their spectral emission lines in the relevant LIBS spectra. In addition, CN and C2 molecular emission bands in the same spectra have been studied as indicators of proteins content in the samples. The obtained analytical results demonstrate the higher elemental contents of the maternal milk compared with the commercial formulas samples. Similar results have been obtained as for the proteins content. It has been also shown that calcium and proteins have similar relative concentration trends in the studied samples. This work demonstrates the feasibility of adopting LIBS as a fast, safe, less costly technique evaluating qualitatively the nutrients content of both maternal and commercial milk samples. Copyright © 2013 Elsevier B.V. All rights reserved.

Use of species-specific PCR for the identification of 10 sea cucumber species

NASA Astrophysics Data System (ADS)

Wen, Jing; Zeng, Ling

2014-11-01

We developed a species-specific PCR method to identify species among dehydrated products of 10 sea cucumber species. Ten reverse species-specific primers designed from the 16S rRNA gene, in combination with one forward universal primer, generated PCR fragments of ca. 270 bp length for each species. The specificity of the PCR assay was tested with DNA of samples of 21 sea cucumber species. Amplification was observed in specific species only. The species-specific PCR method we developed was successfully applied to authenticate species of commercial products of dehydrated sea cucumber, and was proven to be a useful, rapid, and low-cost technique to identify the origin of the sea cucumber product.

Chapter 6. Dwarf mistletoe surveys

Treesearch

J.A. Muir; B. Moody

2002-01-01

Dwarf mistletoe surveys are conducted for a variety of vegetation management objectives. Various survey and sampling techniques are used either at a broad, landscape scale in forest planning or program review, or at an individual, stand, site level for specific project implementation. Standard and special surveys provide data to map mistletoe distributions and quantify...

Development and evaluation of 16S rRNA gene targeting Enterococcus genus- and species-specific assays

EPA Science Inventory

Enterococci have been widely used as indicators of fecal pollution in recreational waters. Most studies enumerate enterococci using culture-based techniques that are time consuming and do not provide information on the identity of enterococci species within a given sample. Althou...

Targeted Magnetic Hyperthermia for Lung Cancer

DTIC Science & Technology

2012-09-01

Despite significant advances in diagnostic techniques an d the disco very of new molecularl y targeted therapies , lung cancer (specifically, non-small...vibrating sample magneto metry) and heating rates. The effect of MH on overall tumor cell kill was determined in A549 cells (NSCLCs) based on the amou nt of

Development and evaluation of a Loop Mediated Isothermal Amplification (LAMP) technique for the detection of hookworm (Necator americanus) infection in fecal samples.

PubMed

Mugambi, Robert Muriuki; Agola, Eric L; Mwangi, Ibrahim N; Kinyua, Johnson; Shiraho, Esther Andia; Mkoji, Gerald M

2015-11-06

Hookworm infection is a major concern in sub-Saharan Africa, particularly in children and pregnant women. Necator americanus and Ancylostoma duodenale are responsible for this condition. Hookworm disease is one of the Neglected tropical diseases (NTDs) that are targeted for elimination through global mass chemotherapy. To support this there is a need for reliable diagnostic tools. The conventional diagnostic test, Kato-Katz that is based on microscopic detection of parasite ova in faecal samples, is not effective due to its low sensitivity that is brought about mainly by non-random distribution of eggs in stool and day to day variation in egg output. It is tedious, cumbersome to perform and requires experience for correct diagnosis. LAMP-based tests are simple, relatively cheap, offer greater sensitivity, specificity than existing tests, have high throughput capability, and are ideal for use at the point of care. We have developed a LAMP diagnostic test for detection of hookworm infection in faecal samples. LAMP relies on auto cycling strand displacement DNA synthesis performed at isothermal temperature by Bst polymerase and a set of 4 specific primers. The primers used in the LAMP assay were based on the second Internal Transcribed Spacer (ITS-2) region and designed using Primer Explorer version 4 Software. The ITS-2 region of the ribosomal gene (rDNA) was identified as a suitable target due to its low mutation rates and substantial differences between species. DNA was extracted directly from human faecal samples, followed by LAMP amplification at isothermal temperature of 63 °C for 1 h. Amplicons were visualized using gel electrophoresis and SYBR green dye. Both specificity and sensitivity of the assay were determined. The LAMP based technique developed was able to detect N. americanus DNA in faecal samples. The assay showed 100 % specificity and no cross-reaction was observed with other helminth parasites (S. mansoni, A. lumbricoides or T. trichiura). The developed LAMP assay was 97 % sensitive and DNA at concentrations as low as 0.4 fg were amplified. The LAMP assay developed is an appropriate diagnostic method for the detection of N. americanus DNA in human stool samples because of its simplicity, low cost, sensitivity, and specificity. It holds great promise as a useful diagnostic tool for use in disease control where infection intensities have been significantly reduced.

[Effect of mechanical grinding of Sphagnum on the structure and physiological state of bacterial communities].

PubMed

Dobrovol'skaya, T G; Golovchenko, A V; Yakushev, A V; Manucharova, N A; Yurchenko, E N

2014-01-01

The microcosm method was used to demonstrate an increase in bacterial numbers and drastic changes in the taxonomic structure of saprotrophic bacteria as a result of mechanical grinding of Sphagnum moss. Ekkrisotrophic agrobacteria predominant in untreated moss were replaced by hydrolytic bacteria. Molecular biological approaches revealed such specific hydrolytic bacteria as Janthinobacterium agaricum and Streptomyces purpurascens among the dominant taxa. The application of kinetic technique for determination of the physiological state of bacteria in situ revealed higher functional diversity of hydrolytic bacteria in ground moss than in untreated samples. A considerable decrease of the C/N ratio in ground samples of living Sphagnum incubated using the microcosm technique indicated decomposition of this substrate.

Multiwavelength absorbance of filter deposits for determination of environmental tobacco smoke and black carbon

NASA Astrophysics Data System (ADS)

Lawless, Phil A.; Rodes, Charles E.; Ensor, David S.

A multiwavelength optical absorption technique has been developed for Teflon filters used for personal exposure sampling with sufficient sensitivity to allow apportionments of environmental tobacco smoke and soot (black) carbon to be made. Measurements on blank filters show that the filter material itself contributes relatively little to the total absorbance and filters from the same lot have similar characteristics; this makes retrospective analysis of filters quite feasible. Using an integrating sphere radiometer and multiple wavelengths to provide specificity, the determination of tobacco smoke and carbon with reasonable accuracy is possible on filters not characterized before exposure. This technique provides a low cost, non-destructive exposure assessment alternative to both standard thermo-gravimetric elemental carbon evaluations on quartz filters and cotinine analyses from urine or saliva samples. The method allows the same sample filter to be used for assessment of mass, carbon, and tobacco smoke without affecting the deposit.

On the Search for Nuclear Resonance Fluorescence Signatures of 235U and 238U above 3 MeV

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

Warren, Glen A.; Caggiano, Joseph A.; Bertozzi, William

Nuclear resonance fluorescence is a physical process that provides an isotope-specific signature that could be used for the identification and characterization of materials. The technique involves the detection of prompt discrete-energy photons emitted from a sample that is exposed to MeV-energy photons. Potential applications of the technique range from detection of high explosives to characterization of special nuclear materials such as 235U. Pacific Northwest National Laboratory and Passport Systems have collaborated to conduct a pair of measurements to search for a nuclear resonance fluorescence response of 235U above 3 MeV and of 238U above 5 MeV using an 8 gmore » sample of highly enriched uranium and a 90 g sample of depleted uranium. No new signatures were observed. The minimum detectable integrated cross section for 235U is presented.« less

On the Search for Nuclear Resonance Fluorescence Signatures of 235U and 238U above 3 MeV

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

Warren, Glen A.; Caggiano, Joseph A.; Bertozzi, William

Abstract–Nuclear resonance fluorescence is a physical process that provides an isotope-specific signature that could be used for the identification and characterization of materials. The technique involves the detection of prompt discrete-energy photons emitted from a sample that is exposed to photons in the MeV energy range. Potential applications of the technique range from detection of high explosives to characterization of special nuclear materials such as 235U. Pacific Northwest National Laboratory and Passport Systems have collaborated to conduct a a pair of measurements to search for a nuclear resonance fluorescence response of 235U above 3 MeV and of 238U above 5more » MeV using an 8 g sample of highly enriched uranium and a 90 g sample of depleted uranium. No new signatures were observed. The minimum detectable integrated cross section for 235U is presented.« less

SRRF: Universal live-cell super-resolution microscopy.

PubMed

Culley, Siân; Tosheva, Kalina L; Matos Pereira, Pedro; Henriques, Ricardo

2018-08-01

Super-resolution microscopy techniques break the diffraction limit of conventional optical microscopy to achieve resolutions approaching tens of nanometres. The major advantage of such techniques is that they provide resolutions close to those obtainable with electron microscopy while maintaining the benefits of light microscopy such as a wide palette of high specificity molecular labels, straightforward sample preparation and live-cell compatibility. Despite this, the application of super-resolution microscopy to dynamic, living samples has thus far been limited and often requires specialised, complex hardware. Here we demonstrate how a novel analytical approach, Super-Resolution Radial Fluctuations (SRRF), is able to make live-cell super-resolution microscopy accessible to a wider range of researchers. We show its applicability to live samples expressing GFP using commercial confocal as well as laser- and LED-based widefield microscopes, with the latter achieving long-term timelapse imaging with minimal photobleaching. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

NIR and UV-vis spectroscopy, artificial nose and tongue: comparison of four fingerprinting techniques for the characterisation of Italian red wines.

PubMed

Casale, M; Oliveri, P; Armanino, C; Lanteri, S; Forina, M

2010-06-04

Four rapid and low-cost vanguard analytical systems (NIR and UV-vis spectroscopy, a headspace-mass based artificial nose and a voltammetric artificial tongue), together with chemometric pattern recognition techniques, were applied and compared in addressing a food authentication problem: the distinction between wine samples from the same Italian oenological region, according to the grape variety. Specifically, 59 certified samples belonging to the Barbera d'Alba and Dolcetto d'Alba appellations and collected from the same vintage (2007) were analysed. The instrumental responses, after proper data pre-processing, were used as fingerprints of the characteristics of the samples: the results from principal component analysis and linear discriminant analysis were discussed, comparing the capability of the four analytical strategies in addressing the problem studied. Copyright 2010 Elsevier B.V. All rights reserved.

Development of a reverse transcription polymerase chain reaction method for yellow fever virus detection.

PubMed

Méndez, María C; Domingo, Cristina; Tenorio, Antonio; Pardo, Lissethe C; Rey, Gloria J; Méndez, Jairo A

2013-09-01

Yellow fever is considered a re-emerging disease and is endemic in tropical regions of Africa and South America. At present, there are no standardized or commercialized kits available for yellow fever virus detection. Therefore, diagnosis must be made by time-consuming routine techniques, and sometimes, the virus or its proteins are not detected. Furthermore, co-circulation with other flaviviruses, including dengue virus, increases the difficulty of diagnosis. To develop a specific reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR-based assay to improve the detection and diagnosis of yellow fever virus using both serum and fresh tissue samples. RT-PCR primers were designed to amplify a short fragment of all yellow fever virus genotypes reported. A second set of primers was used in a nested PCR to increase sensitivity. Thirty-three clinical samples were tested with the standardized reaction. The expected amplicon was obtained in 25 out of 33 samples analyzed using this approach, and 2 more samples tested positive after a subsequent nested PCR approach. This improved technique not only ensures the specific detection of a wide range of yellow fever virus genotypes but also may increase the sensitivity of detection by introducing a second round of amplification, allowing a rapid differential diagnosis between dengue and yellow fever infection, which is required for effective surveillance and opportune epidemiologic measures.

Accuracy of specific BIVA for the assessment of body composition in the United States population.

PubMed

Buffa, Roberto; Saragat, Bruno; Cabras, Stefano; Rinaldi, Andrea C; Marini, Elisabetta

2013-01-01

Bioelectrical impedance vector analysis (BIVA) is a technique for the assessment of hydration and nutritional status, used in the clinical practice. Specific BIVA is an analytical variant, recently proposed for the Italian elderly population, that adjusts bioelectrical values for body geometry. Evaluating the accuracy of specific BIVA in the adult U.S. population, compared to the 'classic' BIVA procedure, using DXA as the reference technique, in order to obtain an interpretative model of body composition. A cross-sectional sample of 1590 adult individuals (836 men and 754 women, 21-49 years old) derived from the NHANES 2003-2004 was considered. Classic and specific BIVA were applied. The sensitivity and specificity in recognizing individuals below the 5(th) and above the 95(th) percentiles of percent fat (FMDXA%) and extracellular/intracellular water (ECW/ICW) ratio were evaluated by receiver operating characteristic (ROC) curves. Classic and specific BIVA results were compared by a probit multiple-regression. Specific BIVA was significantly more accurate than classic BIVA in evaluating FMDXA% (ROC areas: 0.84-0.92 and 0.49-0.61 respectively; p = 0.002). The evaluation of ECW/ICW was accurate (ROC areas between 0.83 and 0.96) and similarly performed by the two procedures (p = 0.829). The accuracy of specific BIVA was similar in the two sexes (p = 0.144) and in FMDXA% and ECW/ICW (p = 0.869). Specific BIVA showed to be an accurate technique. The tolerance ellipses of specific BIVA can be used for evaluating FM% and ECW/ICW in the U.S. adult population.

Specific binding of magnetic nanoparticle probes to platelets in whole blood detected by magnetorelaxometry

NASA Astrophysics Data System (ADS)

Eberbeck, Dietmar; Wiekhorst, Frank; Steinhoff, Uwe; Schwarz, Kay Oliver; Kummrow, Andreas; Kammel, Martin; Neukammer, Jörg; Trahms, Lutz

2009-05-01

The binding of monoclonal antibodies labelled with magnetic nanoparticles to CD61 surface proteins expressed by platelets in whole blood samples was measured by magnetorelaxometry. This technique is sensitive to immobilization of the magnetic labels upon binding. Control experiments with previous saturation of the epitopes on the platelet surfaces demonstrated the specificity of the binding. The kinetics of the antibody antigen reaction is accessible with a temporal resolution of 12 s. The minimal detectable platelet concentration is about 2000 μL -1 (sample volume 150 μL). The proportionality of the magnetic relaxation amplitude to the number of bound labels allows a quantification of the antibody binding capacity.

Quantum-inspired algorithm for estimating the permanent of positive semidefinite matrices

NASA Astrophysics Data System (ADS)

Chakhmakhchyan, L.; Cerf, N. J.; Garcia-Patron, R.

2017-08-01

We construct a quantum-inspired classical algorithm for computing the permanent of Hermitian positive semidefinite matrices by exploiting a connection between these mathematical structures and the boson sampling model. Specifically, the permanent of a Hermitian positive semidefinite matrix can be expressed in terms of the expected value of a random variable, which stands for a specific photon-counting probability when measuring a linear-optically evolved random multimode coherent state. Our algorithm then approximates the matrix permanent from the corresponding sample mean and is shown to run in polynomial time for various sets of Hermitian positive semidefinite matrices, achieving a precision that improves over known techniques. This work illustrates how quantum optics may benefit algorithm development.

A blood tumor marker combination assay produces high sensitivity and specificity for cancer according to the natural history.

PubMed

Kobayashi, Tsuneo

2018-03-01

Diagnosis using a specific tumor marker is difficult because the sensitivity of this detection method is under 20%. Herein, a tumor marker combination assay, combining growth-related tumor marker and associated tumor marker (Cancer, 73(7), 1994), was employed. This double-blind tumor marker combination assay (TMCA) showed 87.5% sensitivity as the results, but a low specificity, ranging from 30 to 76%. To overcome this low specificity, we exploited complex markers, a multivariate analysis and serum fractionation by biochemical biopsy. Thus, in this study, a combination of new techniques was used to re-evaluate these serum samples. Three serum panels, containing 90, 120, and 97 samples were obtained from the Mayo Clinic. The final results showed 80-90% sensitivity, 84-85% specificity, and 83-88% accuracy. We demonstrated a notable tumor marker combination assay with high accuracy. This TMCA should be applicable for primary cancer detection and recurrence prevention. © 2018 The Author. Cancer Medicine published by John Wiley & Sons Ltd.

Can direct gas-liquid chromatography of clinical samples detect specific organisms?

PubMed Central

Watt, B; Geddes, P A; Greenan, O A; Napier, S K; Mitchell, A

1982-01-01

A total of 1929 samples was analyzed by direct gas-liquid chromatography and the volatile fatty acid (VFA) patterns of the positive samples were compared with the results of culture. There was no correlation between any bacterial genus or species and the detailed VFA patterns although the presence of butyric or valeric acids, or both, was generally associated with the presence of anaerobes and that of acetic acid was generally associated with aerobic bacteria; however, the technique could not predict the nature of the subsequent bacterial isolate. There was also poor correlation between the VFA pattern in a given sample and the VFA pattern(s) of anaerobic bacteria subsequently isolated from that sample. PMID:7096589

Sampling methods to the statistical control of the production of blood components.

PubMed

Pereira, Paulo; Seghatchian, Jerard; Caldeira, Beatriz; Santos, Paula; Castro, Rosa; Fernandes, Teresa; Xavier, Sandra; de Sousa, Gracinda; de Almeida E Sousa, João Paulo

2017-12-01

The control of blood components specifications is a requirement generalized in Europe by the European Commission Directives and in the US by the AABB standards. The use of a statistical process control methodology is recommended in the related literature, including the EDQM guideline. The control reliability is dependent of the sampling. However, a correct sampling methodology seems not to be systematically applied. Commonly, the sampling is intended to comply uniquely with the 1% specification to the produced blood components. Nevertheless, on a purely statistical viewpoint, this model could be argued not to be related to a consistent sampling technique. This could be a severe limitation to detect abnormal patterns and to assure that the production has a non-significant probability of producing nonconforming components. This article discusses what is happening in blood establishments. Three statistical methodologies are proposed: simple random sampling, sampling based on the proportion of a finite population, and sampling based on the inspection level. The empirical results demonstrate that these models are practicable in blood establishments contributing to the robustness of sampling and related statistical process control decisions for the purpose they are suggested for. Copyright © 2017 Elsevier Ltd. All rights reserved.

Loop-Mediated Isothermal Amplification Label-Based Gold Nanoparticles Lateral Flow Biosensor for Detection of Enterococcus faecalis and Staphylococcus aureus

PubMed Central

Wang, Yi; Li, Hui; Wang, Yan; Zhang, Lu; Xu, Jianguo; Ye, Changyun

2017-01-01

The report describes a simple, rapid and sensitive assay for visual and multiplex detection of Enterococcus faecalis and Staphylococcus aureus based on multiple loop-mediated isothermal amplification (mLAMP) and lateral flow biosensor (LFB). Detection and differentiation of the Ef0027 gene (E. faecalis-specific gene) and nuc gene (S. aureus-specific gene) were determined using fluorescein (FITC)-and digoxin-modified primers in the mLAMP process. In the presence of biotin- and FITC-/digoxin-modified primers, the mLAMP yielded numerous biotin- and FITC-/digoxin-attached duplex products, which were detected by LFB through biotin/streptavidin interaction (biotin on the duplex and streptavidin on the gold nanoparticle) and immunoreactions (FITC/digoxin on the duplex and anti-FITC/digoxin on the LFB test line). The accumulation of gold nanoparticles generated a characteristic red line, enabling visual and multiplex detection of target pathogens without instrumentation. The limit of detection (LoD), analytical specificity and feasibility of LAMP-LFB technique were successfully examined in pure culture and blood samples. The entire procedure, including specimen (blood samples) processing (30 min), isothermal reaction (40 min) and result reporting (within 2 min), could be completed within 75 min. Thus, this assay offers a simple, rapid, sensitive and specific test for multiplex detection of E. faecalis and S. aureus strains. Furthermore, the LAMP-LFB strategy is a universal technique, which can be extended to detect various target sequences by re-designing the specific LAMP primers. PMID:28239371

Raman spectral analysis for rapid screening of dengue infection

NASA Astrophysics Data System (ADS)

Mahmood, T.; Nawaz, H.; Ditta, A.; Majeed, M. I.; Hanif, M. A.; Rashid, N.; Bhatti, H. N.; Nargis, H. F.; Saleem, M.; Bonnier, F.; Byrne, H. J.

2018-07-01

Infection with the dengue virus is currently clinically detected according to different biomarkers in human blood plasma, commonly measured by enzyme linked immunosorbent assays, including non-structural proteins (Ns1), immunoglobulin M (IgM) and immunoglobulin G (IgG). However, there is little or no mutual correlation between the biomarkers, as demonstrated in this study by a comparison of their levels in samples from 17 patients. As an alternative, the label free, rapid screening technique, Raman spectroscopy has been used for the characterisation/diagnosis of healthy and dengue infected human blood plasma samples. In dengue positive samples, changes in specific Raman spectral bands associated with lipidic and amino acid/protein content are observed and assigned based on literature and these features can be considered as markers associated with dengue development. Based on the spectroscopic analysis of the current, albeit limited, cohort of samples, Principal Components Analysis (PCA) coupled Factorial Discriminant Analysis, yielded values of 97.95% sensitivity and 95.40% specificity for identification of dengue infection. Furthermore, in a comparison of the normal samples to the patient samples which scored low for only one of the biomarker tests, but high or medium for either or both of the other two, PCA-FDA demonstrated a sensitivity of 97.38% and specificity of 86.18%, thus providing an unambiguous screening technology.

Efficient inference of population size histories and locus-specific mutation rates from large-sample genomic variation data.

PubMed

Bhaskar, Anand; Wang, Y X Rachel; Song, Yun S

2015-02-01

With the recent increase in study sample sizes in human genetics, there has been growing interest in inferring historical population demography from genomic variation data. Here, we present an efficient inference method that can scale up to very large samples, with tens or hundreds of thousands of individuals. Specifically, by utilizing analytic results on the expected frequency spectrum under the coalescent and by leveraging the technique of automatic differentiation, which allows us to compute gradients exactly, we develop a very efficient algorithm to infer piecewise-exponential models of the historical effective population size from the distribution of sample allele frequencies. Our method is orders of magnitude faster than previous demographic inference methods based on the frequency spectrum. In addition to inferring demography, our method can also accurately estimate locus-specific mutation rates. We perform extensive validation of our method on simulated data and show that it can accurately infer multiple recent epochs of rapid exponential growth, a signal that is difficult to pick up with small sample sizes. Lastly, we use our method to analyze data from recent sequencing studies, including a large-sample exome-sequencing data set of tens of thousands of individuals assayed at a few hundred genic regions. © 2015 Bhaskar et al.; Published by Cold Spring Harbor Laboratory Press.

Synthetic seismograms from vibracores: A case study in correlating the late quaternary seismic stratigraphy of the New Jersey inner continental shelf

USGS Publications Warehouse

Esker, D.; Sheridan, R.E.; Ashley, G.M.; Waldner, J.S.; Hall, D.W.

1996-01-01

A new technique, using empirical relationships between median grain size and density and velocity to calculate proxy values for density and velocity, avoids many of the problems associated with the use of well logs and shipboard measurements to construct synthetic seismograms. This method was used to groundtruth and correlate across both analog and digital shallow high-resolution seismic data on the New Jersey shelf. Sampling dry vibracores to determine median grain size eliminates the detrimental effects that coring disturbances and preservation variables have on the sediment and water content of the core. The link between seismic response to lithology and bed spacing is more exact. The exact frequency of the field seismic data can be realistically simulated by a 10-20 cm sampling interval of the vibracores. The estimate of the percentage error inherent in this technique, 12% for acoustic impedance and 24% for reflection amplitude, is calculated to one standard deviation and is within a reasonable limit for such a procedure. The synthetic seismograms of two cores, 4-6 m long, were used to correlate specific sedimentary deposits to specific seismic reflection responses. Because this technique is applicable to unconsolidated sediments, it is ideal for upper Pleistocene and Holocene strata. Copyright ?? 1996, SEPM (Society for Sedimentary Geology).

Mid-Award Progress Report for Department of Energy Office of Science Graduate Student Research Program

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

Hoffman, M. K.

The purpose of the work proposed for this study is to investigate the behavior and transport mechanisms for cesium-137 in soils collected from contaminated sites with distinct source release scenarios. More specifically, this study aims to determine with which elements and minerals cesium-137 associates in these various environments to more reliably predict its migration in the subsurface. This will be achieved using a state-of-the-art analysis technique available at Lawrence Livermore National Laboratory (LLNL) known as NanoSIMS. Nano-scale secondary ion mass spectrometry, or NanoSIMS, is a destructive surface analysis technique in which positive secondary ions are generated from the surface ofmore » a sample and then quantified based on their mass-to-charge ratio (m/z) using mass spectrometry. The data collected about the secondary ions can then be used to create isotope-specific spatial maps with a resolution of a few hundred nanometers and depth profiles that show the variation of the secondary ion intensity with sputtering time. This should be an ideal technique for locating cesium-137 in a sample, as cesium is an easily ionized element, meaning the yield of secondary cesium (Cs) ions produced should be high and making the identification of cesium-137 straight forward.« less

Bioimaging of metals in brain tissue by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and metallomics.

PubMed

Becker, J Sabine; Matusch, Andreas; Palm, Christoph; Salber, Dagmar; Morton, Kathryn A; Becker, J Susanne

2010-02-01

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has been developed and established as an emerging technique in the generation of quantitative images of metal distributions in thin tissue sections of brain samples (such as human, rat and mouse brain), with applications in research related to neurodegenerative disorders. A new analytical protocol is described which includes sample preparation by cryo-cutting of thin tissue sections and matrix-matched laboratory standards, mass spectrometric measurements, data acquisition, and quantitative analysis. Specific examples of the bioimaging of metal distributions in normal rodent brains are provided. Differences to the normal were assessed in a Parkinson's disease and a stroke brain model. Furthermore, changes during normal aging were studied. Powerful analytical techniques are also required for the determination and characterization of metal-containing proteins within a large pool of proteins, e.g., after denaturing or non-denaturing electrophoretic separation of proteins in one-dimensional and two-dimensional gels. LA-ICP-MS can be employed to detect metalloproteins in protein bands or spots separated after gel electrophoresis. MALDI-MS can then be used to identify specific metal-containing proteins in these bands or spots. The combination of these techniques is described in the second section.

Polymerase chain reaction in the detection of tumor cells: new approaches in diagnosis and follow-up of patients with thyroid cancer.

PubMed

Bojunga, Jörg; Kusterer, Klaus; Schumm-Draeger, Petra-Maria; Usadel, Klaus-Henning

2002-12-01

Thyroid cancers are the most common endocrine malignancies and are being diagnosed with increasing frequency. In addition to other measures, diagnosis is based on fine-needle aspiration cytology examination. Recently, new assays using reverse transcription-polymerase chain reaction (PCR) are being tested to improve sensitivity and specificity of primary diagnosis and detection of recurrent thyroid cancer. In the preoperative diagnosis of thyroid cancer, several tissue- and/or tumor-specific mRNA have been described and in several cases, a higher sensitivity and specificity could be achieved using molecular techniques compared to conventional methods. In the postoperative follow-up of patients with thyroid cancer, conflicting data have been published and the use of PCR techniques revealed several problems of the molecular approach, which are based on some technical as well as biologic limitations. Despite these problems, which are discussed in detail in this review, molecular techniques may nevertheless improve the sensitivity and accuracy of fine-needle aspiration of thyroid nodules, fine-needle aspiration of metastases, and detection of recurrent disease in peripheral blood samples.

Semiquantitative Dot Blot with the GRA8 antigen to differentiate the stages of toxoplasmosis infection.

PubMed

Costa, Juan Gabriel; Vilariño, María Julia

2018-06-01

In this work we present a novel methodology to differentiate the phases of toxoplasmosis infection: the "semiquantitative Dot Blot". It is a simple technique that does not require expensive equipment, does not involve a long technique development, and can be used in a low-complexity laboratory. In this study, two recombinant sequences of Toxoplasma gondii GRA8 antigen were used, and specific IgG antibodies were detected in selected patient samples. This method makes it possible to obtain a score for each serum and define whether the patient is in the acute or chronic phase of the infection. The sensitivity and specificity results varied depending on the antigenic sequence used. With GRA8A, 62.1% and 72.7% were obtained, while with GRA8B, 82.8% and 72.1% were obtained, respectively. Although the sensitivity and specificity values were not close to 100%, they were similar to those reported with the same antigens in ELISA. Therefore, this quantitative technique would be a good alternative to ELISA. Copyright © 2018 Elsevier B.V. All rights reserved.

Refractive index profilometry using the total internally reflected light field.

PubMed

Das, Tania; Bhattacharya, K

2017-11-20

A full-field polarization-based technique is presented for quantitative evaluation of the spatial distribution of the refractive index in macro and micro samples. The sample is mounted on a glass-air interface of a prism, illuminated by a linearly polarized collimated light beam, and two intensity frames are digitally recorded with specific orientations of an analyzer. The pair of intensity data frames captured with this simple setup is combined through an algorithm specially developed for the purpose, to yield the phase difference between the transverse electric and transverse magnetic components of the total internally reflected light field. The phase difference is then related to the refractive index of the sample. Experimental results for refractive index variations in a laser-etched glass plate and red blood corpuscles are presented. One of the salient features of the proposed technique is that the depth of measurement is dependent on the penetration depth of the sample's evanescent field, which is typically of the order of a few hundred nanometers, thereby facilitating refractive index measurements along a thin section of the sample.

Molecular weight dependent structure and charge transport in MAPLE-deposited poly(3-hexylthiophene) thin films

DOE PAGES

Dong, Ban Xuan; Smith, Mitchell; Strzalka, Joseph; ...

2018-02-06

In this work, poly(3-hexylthiophene) (P3HT) films prepared using the matrix-assisted pulsed laser evaporation (MAPLE) technique are shown to possess morphological structures that are dependent on molecular weight (MW). Specifically, the structures of low MW samples of MAPLE-deposited film are composed of crystallites/aggregates embedded within highly disordered environments, whereas those of high MW samples are composed of aggregated domains connected by long polymer chains. Additionally, the crystallite size along the side-chain (100) direction decreases, whereas the conjugation length increases with increasing molecular weight. This is qualitatively similar to the structure of spin-cast films, though the MAPLE-deposited films are more disordered. In-planemore » carrier mobilities in the MAPLE-deposited samples increase with MW, consistent with the notion that longer chains bridge adjacent aggregated domains thereby facilitating more effective charge transport. The carrier mobilities in the MAPLE-deposited simples are consistently lower than those in the solvent-cast samples for all molecular weights, consistent with the shorter conjugation length in samples prepared by this deposition technique.« less

Molecular weight dependent structure and charge transport in MAPLE-deposited poly(3-hexylthiophene) thin films

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

Dong, Ban Xuan; Smith, Mitchell; Strzalka, Joseph

In this work, poly(3-hexylthiophene) (P3HT) films prepared using the matrix-assisted pulsed laser evaporation (MAPLE) technique are shown to possess morphological structures that are dependent on molecular weight (MW). Specifically, the structures of low MW samples of MAPLE-deposited film are composed of crystallites/aggregates embedded within highly disordered environments, whereas those of high MW samples are composed of aggregated domains connected by long polymer chains. Additionally, the crystallite size along the side-chain (100) direction decreases, whereas the conjugation length increases with increasing molecular weight. This is qualitatively similar to the structure of spin-cast films, though the MAPLE-deposited films are more disordered. In-planemore » carrier mobilities in the MAPLE-deposited samples increase with MW, consistent with the notion that longer chains bridge adjacent aggregated domains thereby facilitating more effective charge transport. The carrier mobilities in the MAPLE-deposited simples are consistently lower than those in the solvent-cast samples for all molecular weights, consistent with the shorter conjugation length in samples prepared by this deposition technique.« less

Early driver fatigue detection from electroencephalography signals using artificial neural networks.

PubMed

King, L M; Nguyen, H T; Lal, S K L

2006-01-01

This paper describes a driver fatigue detection system using an artificial neural network (ANN). Using electroencephalogram (EEG) data sampled from 20 professional truck drivers and 35 non professional drivers, the time domain data are processed into alpha, beta, delta and theta bands and then presented to the neural network to detect the onset of driver fatigue. The neural network uses a training optimization technique called the magnified gradient function (MGF). This technique reduces the time required for training by modifying the standard back propagation (SBP) algorithm. The MGF is shown to classify professional driver fatigue with 81.49% accuracy (80.53% sensitivity, 82.44% specificity) and non-professional driver fatigue with 83.06% accuracy (84.04% sensitivity and 82.08% specificity).

Evaluation of a new rapid diagnostic test for the detection of influenza and RSV.

PubMed

Gómez, Sara; Prieto, Columbiana; Vera, Carmen; R Otero, Joaquín; Folgueira, Lola

2016-05-01

Influenza viruses and respiratory syncytial virus (RSV) can cause an acute respiratory disease that occurs seasonally in epidemic waves. This retrospective study was conducted to evaluate the Sofia(®) Influenza A+B and the Sofia(®) RSV fluorescence immunoassays (FIAs), two novel rapid detection tests (RDTs) for influenza A and B and RSV. Two hundred and nine breath samples were selected from patients with respiratory symptoms determined to be positive/negative for influenza A, influenza B or RSV using one of the reference diagnostic techniques, cell culture and/or RT-PCR (Simplexa™Flu A/B & RSV). The Sofia Influenza A+B FIA was tested on 123 samples (63 from children and 60 from adults) and the Sofia RSV FIA was tested on 86 pediatric samples. Sensitivity and specificity values of both assays were calculated assuming the reference techniques as the gold standard. Sensitivity and specificity values for the Sofia Influenza A+B FIA were 73.1% and 97.8%, respectively. Sensitivity and specificity values for the Sofia RSV FIA were 87.5% and 86.7%, respectively. The sensitivity results obtained for the two assays were considerably higher than those reported for other RDTs. In conclusion, the Sofia Influenza A+B and the Sofia RSV FIAs are appropriate tools for the rapid diagnosis of these viruses. Copyright © 2015 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Development and evaluation of a real-time fluorescent polymerase chain reaction assay for the detection of bovine contaminates in cattle feed.

PubMed

Rensen, Gabriel; Smith, Wayne; Ruzante, Juliana; Sawyer, Mary; Osburn, Bennie; Cullor, James

2005-01-01

A real-time fluorescent polymerase chain reaction assay for detecting prohibited ruminant materials such as bovine meat and bone meal (BMBM) in cattle feed using primers and FRET probes targeting the ruminant specific mitochondrial cytochrome b gene was developed and evaluated on two different types of cattle feed. Common problems involved with PCR based testing of cattle feed include the presence of high levels of PCR inhibitors and the need for certain pre-sample processing techniques in order to perform DNA extractions. We have developed a pre-sample processing technique for extracting DNA from cattle feed which does not require the feed sample to be ground to a fine powder and utilizes materials that are disposed of between samples, thus, reducing the potential of cross contamination. The DNA extraction method utilizes Whatman FTA card technology, is adaptable to high sample throughput analysis and allows for room temperature storage with established archiving of samples of up to 14 years. The Whatman FTA cards are subsequently treated with RNAse and undergo a Chelex-100 extraction (BioRad, Hercules, CA), thus removing potential PCR inhibitors and eluting the DNA from the FTA card for downstream PCR analysis. The detection limit was evaluated over a period of 30 trials on calf starter mix and heifer starter ration feed samples spiked with known concentrations of BMBM. The PCR detection assay detected 0.05% wt/wt BMBM contamination with 100% sensitivity, 100% specificity, and 100% confidence. Concentrations of 0.005% and 0.001% wt/wt BMBM contamination were also detected in both feed types but with varying levels of confidence.

DNA-based species detection capabilities using laser transmission spectroscopy

PubMed Central

Mahon, A. R.; Barnes, M. A.; Li, F.; Egan, S. P.; Tanner, C. E.; Ruggiero, S. T.; Feder, J. L.; Lodge, D. M.

2013-01-01

Early detection of invasive species is critical for effective biocontrol to mitigate potential ecological and economic damage. Laser transmission spectroscopy (LTS) is a powerful solution offering real-time, DNA-based species detection in the field. LTS can measure the size, shape and number of nanoparticles in a solution and was used here to detect size shifts resulting from hybridization of the polymerase chain reaction product to nanoparticles functionalized with species-specific oligonucleotide probes or with the species-specific oligonucleotide probes alone. We carried out a series of DNA detection experiments using the invasive freshwater quagga mussel (Dreissena bugensis) to evaluate the capability of the LTS platform for invasive species detection. Specifically, we tested LTS sensitivity to (i) DNA concentrations of a single target species, (ii) the presence of a target species within a mixed sample of other closely related species, (iii) species-specific functionalized nanoparticles versus species-specific oligonucleotide probes alone, and (iv) amplified DNA fragments versus unamplified genomic DNA. We demonstrate that LTS is a highly sensitive technique for rapid target species detection, with detection limits in the picomolar range, capable of successful identification in multispecies samples containing target and non-target species DNA. These results indicate that the LTS DNA detection platform will be useful for field application of target species. Additionally, we find that LTS detection is effective with species-specific oligonucleotide tags alone or when they are attached to polystyrene nanobeads and with both amplified and unamplified DNA, indicating that the technique may also have versatility for broader applications. PMID:23015524

Development of duplex RT-PCR-ELISA for the simultaneous detection of hepatitis A virus and hepatitis E virus.

PubMed

Tahk, Hongmin; Lee, Min Hwa; Lee, Kang Bum; Cheon, Doo-Sung; Choi, Changsun

2011-07-01

This study aimed to develop a specific and sensitive duplex reverse transcription polymerase chain reaction enzyme-linked immunosorbent assay (duplex RT-PCR-ELISA) for hepatitis A virus (HAV) and hepatitis E virus (HEV). Duplex RT-PCR-ELISA could detect and differentiate HAV and HEV with specific probes. When ELISA technique was used to detect probe-bound RT-PCR products, duplex RT-PCR-ELISA could detect as little as 0.1 ng/μL HAV and HEV from clinical samples. Human norovirus, enterovirus, poliovirus, murine norovirus and feline calicivirus were used for the specificity test; all were negative. Therefore duplex RT-PCR-ELISA can be used for the simultaneous detection of HAV and HEV in contaminated fecal samples. Copyright © 2011 Elsevier B.V. All rights reserved.

A new comprehensive technique of catheterisation, blood sampling, sample preparation and sample analysis by means of high-pressure liquid chromatography for pharmacokinetic studies with estradiol-linked nitrosoureas and their metabolites.

PubMed

Betsch, B; Berger, M R; Spiegelhalder, B

1990-09-01

Estradiol-linked nitrosoureas are offering new perspectives in the antineoplastic chemotherapy of estradiol-receptor positive mammary carcinomas. In such a molecule estradiol has the function of a carrier which brings about a specific accumulation of the anticancer drug in estradiol-receptor containing tumor cells. However, there is only little knowledge about the pharmacokinetic behavior of this new group of anticancer agents. For that reason a new comprehensive technique of catheterisation, blood sampling, sample preparation and sample analysis with high-pressure liquid chromatography (HPLC) for preclinical pharmacokinetic studies with estradiol-linked nitrosoureas and their metabolites has been developed. N-(2-Chloroethyl)-N-nitroso-carbamoyl-L-alanine-estradiol-17-ester (CNC-alanine-estradiol-17-ester) and N-(2-chloroethyl)-N-nitroso-carbamoyl-L-alanine (CNC-alanine) were used as test compounds. The drugs were tested in female Sprague-Dawley rats with chemically induced mammary carcinomas. The laboratory animals were supplied with two catheters prior to the pharmacokinetic experiments. The blood samples were drawn from the vena cava catheter after the drug had been applied through a vena jugularis catheter. The compounds were extracted from plasma with C18 silicagel reversed phase cartridges. The clean-up technique delivered clear samples only slightly contaminated with the biological matrix. The recovery from plasma was 75 +/- 5% for the hormone-linked CNC-alanine-estradiol-17-ester and 70 +/- 5% for the unlinked CNC-alanine. The analysis was carried out by means of HPLC.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of DNA Cytosine Methylation Patterns Using Methylation-Sensitive Amplification Polymorphism (MSAP).

PubMed

Guevara, María Ángeles; de María, Nuria; Sáez-Laguna, Enrique; Vélez, María Dolores; Cervera, María Teresa; Cabezas, José Antonio

2017-01-01

Different molecular techniques have been developed to study either the global level of methylated cytosines or methylation at specific gene sequences. One of them is the methylation-sensitive amplified polymorphism technique (MSAP) which is a modification of amplified fragment length polymorphism (AFLP). It has been used to study methylation of anonymous CCGG sequences in different fungi, plants, and animal species. The main variation of this technique resides on the use of isoschizomers with different methylation sensitivity (such as HpaII and MspI) as a frequent-cutter restriction enzyme. For each sample, MSAP analysis is performed using both EcoRI/HpaII- and EcoRI/MspI-digested samples. A comparative analysis between EcoRI/HpaII and EcoRI/MspI fragment patterns allows the identification of two types of polymorphisms: (1) methylation-insensitive polymorphisms that show common EcoRI/HpaII and EcoRI/MspI patterns but are detected as polymorphic amplified fragments among samples and (2) methylation-sensitive polymorphisms which are associated with the amplified fragments that differ in their presence or absence or in their intensity between EcoRI/HpaII and EcoRI/MspI patterns. This chapter describes a detailed protocol of this technique and discusses the modifications that can be applied to adjust the technology to different species of interest.

Comparative study of inorganic elements determined in whole blood from Dmd(mdx)/J mice strain by EDXRF and NAA analytical techniques.

PubMed

Redígolo, M M; Sato, I M; Metairon, S; Zamboni, C B

2016-04-01

Several diseases can be diagnosed observing the variation of specific elements concentration in body fluids. In this study the concentration of inorganic elements in blood samples of dystrophic (Dmd(mdx)/J) and C57BL/6J (control group) mice strain were determined. The results obtained from Energy Dispersive X-ray Fluorescence (EDXRF) were compared with Neutron Activation Analysis (NAA) technique. Both analytical techniques showed to be appropriate and complementary offering a new contribution for veterinary medicine as well as detailed knowledge of this pathology. Copyright © 2016 Elsevier Ltd. All rights reserved.

As-built design specification for proportion estimate software subsystem

NASA Technical Reports Server (NTRS)

Obrien, S. (Principal Investigator)

1980-01-01

The Proportion Estimate Processor evaluates four estimation techniques in order to get an improved estimate of the proportion of a scene that is planted in a selected crop. The four techniques to be evaluated were provided by the techniques development section and are: (1) random sampling; (2) proportional allocation, relative count estimate; (3) proportional allocation, Bayesian estimate; and (4) sequential Bayesian allocation. The user is given two options for computation of the estimated mean square error. These are referred to as the cluster calculation option and the segment calculation option. The software for the Proportion Estimate Processor is operational on the IBM 3031 computer.

Comparison of streamflow and water-quality data collection techniques for the Saginaw River, Michigan

USGS Publications Warehouse

Hoard, C.J.; Holtschlag, D.J.; Duris, J.W.; James, D.A.; Obenauer, D.J.

2012-01-01

In 2009, the Michigan Department of Environmental Quality and the U.S. Geological Survey developed a plan to compare the effect of various streamgaging and water-quality collection techniques on streamflow and stream water-quality data for the Saginaw River, Michigan. The Saginaw River is the primary contributor of surface runoff to Saginaw Bay, Lake Huron, draining approximately 70 percent of the Saginaw Bay watershed. The U.S. Environmental Protection Agency has listed the Saginaw Bay system as an "Area of Concern" due to many factors, including excessive sediment and nutrient concentrations in the water. Current efforts to estimate loading of sediment and nutrients to Saginaw Bay utilize water-quality samples collected using a surface-grab technique and flow data that are uncertain during specific conditions. Comparisons of current flow and water-quality sampling techniques to alternative techniques were assessed between April 2009 and September 2009 at two locations in the Saginaw River. Streamflow estimated using acoustic Doppler current profiling technology was compared to a traditional stage-discharge technique. Complex conditions resulting from the influence of Saginaw Bay on the Saginaw River were able to be captured using the acoustic technology, while the traditional stage-discharge technique failed to quantify these effects. Water-quality samples were collected at two locations and on eight different dates, utilizing both surface-grab and depth-integrating multiple-vertical techniques. Sixteen paired samples were collected and analyzed for suspended sediment, turbidity, total phosphorus, total nitrogen, orthophosphate, nitrite, nitrate, and ammonia. Results indicate that concentrations of constituents associated with suspended material, such as suspended sediment, turbidity, and total phosphorus, are underestimated when samples are collected using the surface-grab technique. The median magnitude of the relative percent difference in concentration based on sampling technique was 37 percent for suspended sediment, 26 percent for turbidity, and 9.7 percent for total phosphorus samples collected at both. Acoustic techniques were also used to assist in the determination of the effectiveness of using acoustic-backscatter information for estimating the suspended-sediment concentration of the river water. Backscatter data was collected by use of an acoustic Doppler current profiler, and a Van Dorn manual sampler was simultaneously used to collect discrete water samples at 10 depths (3.5, 7.5, 11, 14, 15.5, 17.5, 19.5, 20.5, 22, and 24.5 ft below the water surface) along two vertical profiles near the center of the Saginaw River near Bay City. The Van Dorn samples were analyzed for suspended-sediment concentrations, and these data were then used to develop a relationship between acoustic-backscatter data. Acoustic-backscatter data was strongly correlated to sediment concentrations and, by using a linear regression, was able to explain 89 percent of the variability. Although this regression technique showed promise for using acoustic backscatter to estimate suspended-sediment concentration, attempts to compare suspended-sediment concentrations to the acoustic signal-to-noise ratio estimates, recorded at the fixed acoustic streamflow-gaging station near Bay City (04157061), resulted in a poor correlation.

Bioelectrographic testing of mineral samples: a comparison of techniques.

PubMed

Vainshelboim, Alex; Momoh, Kenneth S

2005-04-01

This study was initiated to determine the suitability of differing techniques to record optical properties of gemstones under electromagnetic stimulation. Such properties are of interest due to the historical use of gemstones in folkloric remedies, specifically as agents for concentrating, focusing, or otherwise conducting energy flows in the human body. The techniques researched produce a localized corona discharge around the tested material. The simplest technique, Tesla coil Kirlian photography (TCKP), uses a Tesla coil to introduce a strong electric current, and the circuit is completed by a glass electrode. The corona discharge is then photographed. The other technique used in the study is gas discharge visualization (GDV), which uses a pulsed current and a digital camera integral to the coil to produce digital images of the corona discharge. Gemstones were tested both whole and in powdered form. The sample gemstones were amethyst, aquamarine, garnet, golden citrine, pink tourmaline, and yellow topaz. Powdered gemstones were ground to a particle size of 2-5 microns; whole gemstones were roundcut to a diameter of 5 mm. In our tests, TCKP showed divergent effects for differing types of gemstone. The most extreme effects were exhibited by tourmaline, both in powdered and whole form. In addition, TCKP appeared to indicate differing effects for gemstones of the identical type but mined from differing locations. The GDV technique showed differing data among the gemstones for the measured parameters, indicating that a high relative intensity did not correspond to the size of the corona discharge. While both techniques showed promise in distinguishing differences in corona discharge behavior in gemstone samples, further work is necessary to determine the significance of differences in geographical sources or between gemstones of similar crystalline structure. The techniques explored show promise in characterizing the properties of gem materials under electromagnetic stimulation.

The measurement of linear frequency drift in oscillators

NASA Astrophysics Data System (ADS)

Barnes, J. A.

1985-04-01

A linear drift in frequency is an important element in most stochastic models of oscillator performance. Quartz crystal oscillators often have drifts in excess of a part in ten to the tenth power per day. Even commercial cesium beam devices often show drifts of a few parts in ten to the thirteenth per year. There are many ways to estimate the drift rates from data samples (e.g., regress the phase on a quadratic; regress the frequency on a linear; compute the simple mean of the first difference of frequency; use Kalman filters with a drift term as one element in the state vector; and others). Although most of these estimators are unbiased, they vary in efficiency (i.e., confidence intervals). Further, the estimation of confidence intervals using the standard analysis of variance (typically associated with the specific estimating technique) can give amazingly optimistic results. The source of these problems is not an error in, say, the regressions techniques, but rather the problems arise from correlations within the residuals. That is, the oscillator model is often not consistent with constraints on the analysis technique or, in other words, some specific analysis techniques are often inappropriate for the task at hand. The appropriateness of a specific analysis technique is critically dependent on the oscillator model and can often be checked with a simple whiteness test on the residuals.

Selective parathyroid venous sampling in primary hyperparathyroidism: A systematic review and meta-analysis.

PubMed

Ibraheem, Kareem; Toraih, Eman A; Haddad, Antoine B; Farag, Mahmoud; Randolph, Gregory W; Kandil, Emad

2018-05-14

Minimally invasive parathyroidectomy requires accurate preoperative localization techniques. There is considerable controversy about the effectiveness of selective parathyroid venous sampling (sPVS) in primary hyperparathyroidism (PHPT) patients. The aim of this meta-analysis is to examine the diagnostic accuracy of sPVS as a preoperative localization modality in PHPT. Studies evaluating the diagnostic accuracy of sPVS for PHPT were electronically searched in the PubMed, EMBASE, Web of Science, and Cochrane Controlled Trials Register databases. Two independent authors reviewed the studies, and revised quality assessment of diagnostic accuracy study tool was used for the quality assessment. Study heterogeneity and pooled estimates were calculated. Two hundred and two unique studies were identified. Of those, 12 studies were included in the meta-analysis. Pooled sensitivity, specificity, and positive likelihood ratio (PLR) of sPVS were 74%, 41%, and 1.55, respectively. The area-under-the-receiver operating characteristic curve was 0.684, indicating an average discriminatory ability of sPVS. On comparison between sPVS and noninvasive imaging modalities, sensitivity, PLR, and positive posttest probability were significantly higher in sPVS compared to noninvasive imaging modalities. Interestingly, super-selective venous sampling had the highest sensitivity, accuracy, and positive posttest probability compared to other parathyroid venous sampling techniques. This is the first meta-analysis to examine the accuracy of sPVS in PHPT. sPVS had higher pooled sensitivity when compared to noninvasive modalities in revision parathyroid surgery. However, the invasiveness of this technique does not favor its routine use for preoperative localization. Super-selective venous sampling was the most accurate among all other parathyroid venous sampling techniques. Laryngoscope, 2018. © 2018 The American Laryngological, Rhinological and Otological Society, Inc.

Efficient and Specific Detection of Salmonella in Food Samples Using a stn-Based Loop-Mediated Isothermal Amplification Method

PubMed Central

2015-01-01

The Salmonella enterotoxin (stn) gene exhibits high homology among S. enterica serovars and S. bongori. A set of 6 specific primers targeting the stn gene were designed for detection of Salmonella spp. using the loop-mediated isothermal amplification (LAMP) method. The primers amplified target sequences in all 102 strains of 87 serovars of Salmonella tested and no products were detected in 57 non-Salmonella strains. The detection limit in pure cultures was 5 fg DNA/reaction when amplified at 65°C for 25 min. The LAMP assay could detect Salmonella in artificially contaminated food samples as low as 220 cells/g of food without a preenrichment step. However, the sensitivity was increased 100-fold (~2 cells/g) following 5 hr preenrichment at 35°C. The LAMP technique, with a preenrichment step for 5 and 16 hr, was shown to give 100% specificity with food samples compared to the reference culture method in which 67 out of 90 food samples gave positive results. Different food matrixes did not interfere with LAMP detection which employed a simple boiling method for DNA template preparation. The results indicate that the LAMP method, targeting the stn gene, has great potential for detection of Salmonella in food samples with both high specificity and high sensitivity. PMID:26543859

Evaluation of a New Technique for iFOBT Utilising a New Sample Collection Device with Increased Buffer Stability.

PubMed

Bruns-Toepler, Markus; Hardt, Philip

2017-07-01

The aims of the present study were: (i) Evaluate specificity and sensitivity of Hb Smart enzyme-linked immunosorbent assay (ELISA) (ScheBo Biotech) compared to colonoscopy results and (ii) assess stability of a new sample collection device containing a newly formulated buffer to extract haemoglobin using buffer and stool samples spiked with defined concentrations of haemoglobin. Stool samples were quantified with the ELISA method. The stability of haemoglobin in the extraction buffer and in native stool samples, respectively, was determined daily by ELISA during storage for 5 days at 4°C and at room temperature after addition of haemoglobin. Haemoglobin ELISA had a sensitivity of 78.4% for detection of CRC with a specificity of 98%. Haemoglobin extracted in corresponding extraction buffer demonstrated stability throughout storage for 5 days at 4°C and at room temperature. Hb Smart represents a very promising tool for large-scale screening of CRC with regard to sample handling, stability and analysis of haemoglobin in faeces. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Nucleic acid probes as a diagnostic method for tick-borne hemoparasites of veterinary importance.

PubMed

Figueroa, J V; Buening, G M

1995-03-01

An increased number of articles on the use of nucleic acid-based hybridization techniques for diagnostic purposes have been recently published. This article reviews nucleic acid-based hybridization as an assay to detect hemoparasite infections of economic relevance in veterinary medicine. By using recombinant DNA techniques, selected clones containing inserts of Anaplasma, Babesia, Cowdria or Theileria genomic DNA sequences have been obtained, and they are now available to be utilized as specific, highly sensitive DNA or RNA probes to detect the presence of the hemoparasite DNA in an infected animal. Either in an isotopic or non-isotopic detection system, probes have allowed scientists to test for--originally in samples collected from experimentally infected animals and later in samples collected in the field--the presence of hemoparasites during the prepatent, patent, convalescent, and chronic periods of the infection in the host. Nucleic acid probes have given researchers the opportunity to carry out genomic analysis of parasite DNA to differentiate hemoparasite species and to identify genetically distinct populations among and within isolates, strains and clonal populations. Prevalence of parasite infection in the tick vector can now be accomplished more specifically with the nucleic acid probes. Lately, with the advent of the polymerase chain reaction technique, small numbers of hemoparasites can be positively identified in the vertebrate host and tick vector. These techniques can be used to assess the veterinary epidemiological situation in a particular geographical region for the planning of control measures.

Cell purification: a new challenge for biobanks.

PubMed

Almeida, Maria; García-Montero, Andres C; Orfao, Alberto

2014-01-01

Performing '-omics' analyses on heterogeneous biological tissue samples, such as blood or bone marrow, can lead to biased or even erroneous results, particularly when the targeted cells and/or molecules are present at relatively low percentages/amounts. In such cases, whole sample analysis will most probably dilute and mask the features of the cell and/or molecules of interest, and this will negatively impact the results and their interpretation. Therefore, frequently it is critically important to have well-characterized and high-quality purified cell populations for the reliable detection of subtle variations in their specific features, such as gene expression profile, protein expression pattern and metabolic status. Biobanks are technological platforms which aim to provide researchers access to a large number of high-quality biological samples and their associated data, particularly to support high-quality scientific and clinical research projects, and such projects will benefit enormously by having access to high-quality purified cell populations or their biological components (e.g. DNA, RNA, proteins). Therefore, a clear opportunity exists for preparative cell sorting techniques in biobanks. Although multiple different cell purification approaches exist or are under development (e.g. cell purification techniques based on cell adherence, density and/or cell size properties, methods based on antibody binding as well as new lab-on-a-chip purification techniques), the choice for a specific technology depends on multiple variables, including cell recovery, purity and yield, among others. In addition, most cell purification approaches are not well suited for high-throughput (HT) purification of multiple cell populations coexisting in a sample. Here we review the most (currently) used cell sorting methods that may be applied for sample preparation in biobanks. For the different approaches, technical considerations about their advantages and limitations are highlighted, and the requirements to be met by a HT cell sorting technology to be used in biobanks are also discussed.

Locating Stardust-like Particles in Aerogel Using X-Ray Techniques

NASA Technical Reports Server (NTRS)

Jurewicz, A. J. G.; Jones, S. M.; Tsapin, A.; Mih, D. T.; Connolly, H. C., Jr.; Graham, G. A.

2003-01-01

Silica aerogel is the material that the spacecraft STARDUST is using to collect interstellar and cometary silicates. Anticipating the return of the samples to earth in January of 2006, MANY individual investigators and, especially, the investigators in NASA's SRLIDAP program are studying means of both in situ analysis of particles, as well as particle extraction. To help individual PI's with extraction of particles from aerogel in their own laboratories, we are exploring the use of standard laboratory x-ray equipment and commercial techniques for precisely locating specific particles in aerogel. We approached the evaluation of commercial x-ray techniques as follows. First, we determined the most appropriate detector for use with aerogel and particulates. Then, we compared and contrasted techniques useful for university laboratories.

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.

An evaluation of methods for estimating decadal stream loads

NASA Astrophysics Data System (ADS)

Lee, Casey J.; Hirsch, Robert M.; Schwarz, Gregory E.; Holtschlag, David J.; Preston, Stephen D.; Crawford, Charles G.; Vecchia, Aldo V.

2016-11-01

Effective management of water resources requires accurate information on the mass, or load of water-quality constituents transported from upstream watersheds to downstream receiving waters. Despite this need, no single method has been shown to consistently provide accurate load estimates among different water-quality constituents, sampling sites, and sampling regimes. We evaluate the accuracy of several load estimation methods across a broad range of sampling and environmental conditions. This analysis uses random sub-samples drawn from temporally-dense data sets of total nitrogen, total phosphorus, nitrate, and suspended-sediment concentration, and includes measurements of specific conductance which was used as a surrogate for dissolved solids concentration. Methods considered include linear interpolation and ratio estimators, regression-based methods historically employed by the U.S. Geological Survey, and newer flexible techniques including Weighted Regressions on Time, Season, and Discharge (WRTDS) and a generalized non-linear additive model. No single method is identified to have the greatest accuracy across all constituents, sites, and sampling scenarios. Most methods provide accurate estimates of specific conductance (used as a surrogate for total dissolved solids or specific major ions) and total nitrogen - lower accuracy is observed for the estimation of nitrate, total phosphorus and suspended sediment loads. Methods that allow for flexibility in the relation between concentration and flow conditions, specifically Beale's ratio estimator and WRTDS, exhibit greater estimation accuracy and lower bias. Evaluation of methods across simulated sampling scenarios indicate that (1) high-flow sampling is necessary to produce accurate load estimates, (2) extrapolation of sample data through time or across more extreme flow conditions reduces load estimate accuracy, and (3) WRTDS and methods that use a Kalman filter or smoothing to correct for departures between individual modeled and observed values benefit most from more frequent water-quality sampling.

An evaluation of methods for estimating decadal stream loads

USGS Publications Warehouse

Lee, Casey; Hirsch, Robert M.; Schwarz, Gregory E.; Holtschlag, David J.; Preston, Stephen D.; Crawford, Charles G.; Vecchia, Aldo V.

2016-01-01

Effective management of water resources requires accurate information on the mass, or load of water-quality constituents transported from upstream watersheds to downstream receiving waters. Despite this need, no single method has been shown to consistently provide accurate load estimates among different water-quality constituents, sampling sites, and sampling regimes. We evaluate the accuracy of several load estimation methods across a broad range of sampling and environmental conditions. This analysis uses random sub-samples drawn from temporally-dense data sets of total nitrogen, total phosphorus, nitrate, and suspended-sediment concentration, and includes measurements of specific conductance which was used as a surrogate for dissolved solids concentration. Methods considered include linear interpolation and ratio estimators, regression-based methods historically employed by the U.S. Geological Survey, and newer flexible techniques including Weighted Regressions on Time, Season, and Discharge (WRTDS) and a generalized non-linear additive model. No single method is identified to have the greatest accuracy across all constituents, sites, and sampling scenarios. Most methods provide accurate estimates of specific conductance (used as a surrogate for total dissolved solids or specific major ions) and total nitrogen – lower accuracy is observed for the estimation of nitrate, total phosphorus and suspended sediment loads. Methods that allow for flexibility in the relation between concentration and flow conditions, specifically Beale’s ratio estimator and WRTDS, exhibit greater estimation accuracy and lower bias. Evaluation of methods across simulated sampling scenarios indicate that (1) high-flow sampling is necessary to produce accurate load estimates, (2) extrapolation of sample data through time or across more extreme flow conditions reduces load estimate accuracy, and (3) WRTDS and methods that use a Kalman filter or smoothing to correct for departures between individual modeled and observed values benefit most from more frequent water-quality sampling.

Evidence for Coordination and Redox Changes of Iron in Shocked Feldspar from Synchrotron MicroXANES

NASA Technical Reports Server (NTRS)

Delaney, J. S.; Dyar, M. D.; Hoerz, F.; Johnson, J. R.

2003-01-01

Shock modification of feldspar has been documented and experimentally reproduced in many studies since the recognition of maskelynite in Shergotty. Experimentally shocked feldspar samples have been well studied using chemical and crystallographic techniques. The crystallographic, site-specific characterization of major and minor elements is less well documented. We present early x-ray absorption (XAS) spectral data for a suite of albitite samples that were experimentally shocked at pressures between 17 and 50 Gpa.

Three-dimensional visualization of the microvasculature of bile duct ligation-induced liver fibrosis in rats by x-ray phase-contrast imaging computed tomography

NASA Astrophysics Data System (ADS)

Xuan, Ruijiao; Zhao, Xinyan; Hu, Doudou; Jian, Jianbo; Wang, Tailing; Hu, Chunhong

2015-07-01

X-ray phase-contrast imaging (PCI) can substantially enhance contrast, and is particularly useful in differentiating biological soft tissues with small density differences. Combined with computed tomography (CT), PCI-CT enables the acquisition of accurate microstructures inside biological samples. In this study, liver microvasculature was visualized without contrast agents in vitro with PCI-CT using liver fibrosis samples induced by bile duct ligation (BDL) in rats. The histological section examination confirmed the correspondence of CT images with the microvascular morphology of the samples. By means of the PCI-CT and three-dimensional (3D) visualization technique, 3D microvascular structures in samples from different stages of liver fibrosis were clearly revealed. Different types of blood vessels, including portal veins and hepatic veins, in addition to ductular proliferation and bile ducts, could be distinguished with good sensitivity, excellent specificity and excellent accuracy. The study showed that PCI-CT could assess the morphological changes in liver microvasculature that result from fibrosis and allow characterization of the anatomical and pathological features of the microvasculature. With further development of PCI-CT technique, it may become a novel noninvasive imaging technique for the auxiliary analysis of liver fibrosis.

A modified method using the SonoPrep ultrasonic skin permeation system for sampling human interstitial fluid is compatible with proteomic techniques.

PubMed

Lecomte, Marie M J; Atkinson, Kelly R; Kay, Daniel P; Simons, Joanne L; Ingram, John R

2013-02-01

The use of biomarkers in skin is a novel diagnostic tool. Interstitial fluid (ISF) from skin provides a snapshot of proteins secreted at the time of sampling giving insights into the patient's health status. A minimally invasive technique for the transdermal collection of human ISF proteins. A low frequency ultrasonic skin permeation device (SonoPrep ultrasonic skin permeation system) was used to produce micropores in the stratum corneum through which ISF was extracted using a portable pulsed vacuum ISF collection device. On average, protein concentrations recovered ranged between 0.064 and 4.792 μg/μL (mean 1.258 μg/μL). Two-dimensional gel electrophoresis revealed that this sample type was amenable to this type of analysis. Gel images indicated that both highly abundant proteins and lower abundance proteins were isolated from the skin. Western blot analysis confirmed the presence of proteins commonly found in plasma and the epidermis. A minimally invasive method for the transdermal recovery of ISF proteins has been developed. We have demonstrated that ISF samples obtained using this approach can be analysed with proteomic techniques, such as two-dimensional gel electrophoresis and western blots, providing another tool for the identification of disease specific protein biomarkers. © 2012 John Wiley & Sons A/S.

Evaluation of elemental profiling methods, including laser-induced breakdown spectroscopy (LIBS), for the differentiation of Cannabis plant material grown in different nutrient solutions.

PubMed

El-Deftar, Moteaa M; Robertson, James; Foster, Simon; Lennard, Chris

2015-06-01

Laser-induced breakdown spectroscopy (LIBS) is an emerging atomic emission based solid sampling technique that has many potential forensic applications. In this study, the analytical performance of LIBS, as well as that of inductively coupled plasma mass spectrometry (ICP-MS), laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and X-ray microfluorescence (μXRF), was evaluated for the ability to conduct elemental analyses on Cannabis plant material, with a specific investigation of the possible links between hydroponic nutrients and elemental profiles from associated plant material. No such study has been previously published in the literature. Good correlation among the four techniques was observed when the concentrations or peak areas of the elements of interest were monitored. For Cannabis samples collected at the same growth time, the elemental profiles could be related to the use of particular commercial nutrients. In addition, the study demonstrated that ICP-MS, LA-ICP-MS and LIBS are suitable techniques for the comparison of Cannabis samples from different sources, with high discriminating powers being achieved. On the other hand, μXRF method was not suitable for the discrimination of Cannabis samples originating from different growth nutrients. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Visualizing Ebolavirus Particles Using Single-Particle Interferometric Reflectance Imaging Sensor (SP-IRIS).

PubMed

Carter, Erik P; Seymour, Elif Ç; Scherr, Steven M; Daaboul, George G; Freedman, David S; Selim Ünlü, M; Connor, John H

2017-01-01

This chapter describes an approach for the label-free imaging and quantification of intact Ebola virus (EBOV) and EBOV viruslike particles (VLPs) using a light microscopy technique. In this technique, individual virus particles are captured onto a silicon chip that has been printed with spots of virus-specific capture antibodies. These captured virions are then detected using an optical approach called interference reflectance imaging. This approach allows for the detection of each virus particle that is captured on an antibody spot and can resolve the filamentous structure of EBOV VLPs without the need for electron microscopy. Capture of VLPs and virions can be done from a variety of sample types ranging from tissue culture medium to blood. The technique also allows automated quantitative analysis of the number of virions captured. This can be used to identify the virus concentration in an unknown sample. In addition, this technique offers the opportunity to easily image virions captured from native solutions without the need for additional labeling approaches while offering a means of assessing the range of particle sizes and morphologies in a quantitative manner.

Sensitive determination of polycyclic aromatic hydrocarbons in water samples by HPLC coupled with SPE based on graphene functionalized with triethoxysilane.

PubMed

Huang, Ke-Jing; Li, Jing; Liu, Yan-Ming; Wang, Lan

2013-02-01

The graphene functionalized with (3-aminopropyl) triethoxysilane was synthesized by a simple hydrothermal reaction and applied as SPE sorbents to extract trace polycyclic aromatic hydrocarbons (PAHs) from environmental water samples. These sorbents possess high adsorption capacity and extraction efficiency due to strong adsorption ability of carbon materials and large specific surface area of nanoparticles, and only 10 mg of sorbents are required to extract PAHs from 100 mL water samples. Several condition parameters, such as eluent and its volume, adsorbent amount, sample volume, sample pH, and sample flow rate, were optimized to achieve good sensitivity and precision. Under the optimized extraction conditions, the method showed good linearity in the range of 1-100 μg/L, repeatability of the extraction (the RSDs were between 1.8 and 2.9%, n = 6), and satisfactory detection limits of 0.029-0.1 μg/L. The recoveries of PAHs spiked in environmental water samples ranged from 84.6 to 109.5%. All these results demonstrated that this new SPE technique was a viable alternative to conventional enrichment techniques for the extraction and analysis of PAHs in complex samples. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Attosecond transient absorption instrumentation for thin film materials: Phase transitions, heat dissipation, signal stabilization, timing correction, and rapid sample rotation.

PubMed

Jager, Marieke F; Ott, Christian; Kaplan, Christopher J; Kraus, Peter M; Neumark, Daniel M; Leone, Stephen R

2018-01-01

We present an extreme ultraviolet (XUV) transient absorption apparatus tailored to attosecond and femtosecond measurements on bulk solid-state thin-film samples, specifically when the sample dynamics are sensitive to heating effects. The setup combines methodology for stabilizing sub-femtosecond time-resolution measurements over 48 h and techniques for mitigating heat buildup in temperature-dependent samples. Single-point beam stabilization in pump and probe arms and periodic time-zero reference measurements are described for accurate timing and stabilization. A hollow-shaft motor configuration for rapid sample rotation, raster scanning capability, and additional diagnostics are described for heat mitigation. Heat transfer simulations performed using a finite element analysis allow comparison of sample rotation and traditional raster scanning techniques for 100 Hz pulsed laser measurements on vanadium dioxide, a material that undergoes an insulator-to-metal transition at a modest temperature of 340 K. Experimental results are presented confirming that the vanadium dioxide (VO 2 ) sample cannot cool below its phase transition temperature between laser pulses without rapid rotation, in agreement with the simulations. The findings indicate the stringent conditions required to perform rigorous broadband XUV time-resolved absorption measurements on bulk solid-state samples, particularly those with temperature sensitivity, and elucidate a clear methodology to perform them.

Attosecond transient absorption instrumentation for thin film materials: Phase transitions, heat dissipation, signal stabilization, timing correction, and rapid sample rotation

NASA Astrophysics Data System (ADS)

Jager, Marieke F.; Ott, Christian; Kaplan, Christopher J.; Kraus, Peter M.; Neumark, Daniel M.; Leone, Stephen R.

2018-01-01

We present an extreme ultraviolet (XUV) transient absorption apparatus tailored to attosecond and femtosecond measurements on bulk solid-state thin-film samples, specifically when the sample dynamics are sensitive to heating effects. The setup combines methodology for stabilizing sub-femtosecond time-resolution measurements over 48 h and techniques for mitigating heat buildup in temperature-dependent samples. Single-point beam stabilization in pump and probe arms and periodic time-zero reference measurements are described for accurate timing and stabilization. A hollow-shaft motor configuration for rapid sample rotation, raster scanning capability, and additional diagnostics are described for heat mitigation. Heat transfer simulations performed using a finite element analysis allow comparison of sample rotation and traditional raster scanning techniques for 100 Hz pulsed laser measurements on vanadium dioxide, a material that undergoes an insulator-to-metal transition at a modest temperature of 340 K. Experimental results are presented confirming that the vanadium dioxide (VO2) sample cannot cool below its phase transition temperature between laser pulses without rapid rotation, in agreement with the simulations. The findings indicate the stringent conditions required to perform rigorous broadband XUV time-resolved absorption measurements on bulk solid-state samples, particularly those with temperature sensitivity, and elucidate a clear methodology to perform them.

Histochemical detection of lead and zinc in plant tissues

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

Tung, G.; Temple, P.J.

1975-01-01

Histochemical studies on uptake and localization of lead and zinc in plant tissues were carried out. A histochemical stain technique was developed to differentiate zinc from lead. Lead was detected in plant tissues by soaking fresh plant materials in freshly prepared sodium rhodizonate stain (0.2% Na rhodizonate acidified to pH3 with glacial acetic acid). Samples were evacuated 5 min and soaked for 30 min before embedding in the congealed stain, then sectioned with a cryostat and examined under a light microscope. Lead particles in plant tissues were stained scarlet-red. Gelatinous, proteinaceous or saccharic embedding materials normally used to prepare plantmore » sampled for sectioning in the cryostat interfered with the color reaction. Sectioning plant samples without staining whole tissues resulted in a weakened response to the stain. Color of stained sample materials were retained for several months if stored in a frozen condition. This technique was used to detect lead both inside and on the surface of plant samples collected in the vicinity of highway and industrial lead sources and to trace the pathways of lead uptake from the air or from contaminated soils. A sodium rhodizonate technique was also developed to be specific for zinc in plant tissues. Plant samples were soaked in a neutral Na-rhodizonate in phosphate buffer at pH 7.5 for observation. The color of zinc developed to produce a purplish or reddish-brown color.« less

In-vivo third-harmonic generation microscopy at 1550nm three-dimensional long-term time-lapse studies in living C. elegans embryos

NASA Astrophysics Data System (ADS)

Aviles-Espinosa, Rodrigo; Santos, Susana I. C. O.; Brodschelm, Andreas; Kaenders, Wilhelm G.; Alonso-Ortega, Cesar; Artigas, David; Loza-Alvarez, Pablo

2011-03-01

In-vivo microscopic long term time-lapse studies require controlled imaging conditions to preserve sample viability. Therefore it is crucial to meet specific exposure conditions as these may limit the applicability of established techniques. In this work we demonstrate the use of third harmonic generation (THG) microscopy for long term time-lapse three-dimensional studies (4D) in living Caenorhabditis elegans embryos employing a 1550 nm femtosecond fiber laser. We take advantage of the fact that THG only requires the existence of interfaces to generate signal or a change in the refractive index or in the χ3 nonlinear coefficient, therefore no markers are required. In addition, by using this wavelength the emitted THG signal is generated at visible wavelengths (516 nm) enabling the use of standard collection optics and detectors operating near their maximum efficiency. This enables the reduction of the incident light intensity at the sample plane allowing to image the sample for several hours. THG signal is obtained through all embryo development stages, providing different tissue/structure information. By means of control samples, we demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Certainly, this technique reduces the complexity of sample preparation (i.e. genetic modification) required by established linear and nonlinear fluorescence based techniques. We demonstrate the non-invasiveness, reduced specimen interference, and strong potential of this particular wavelength to be used to perform long-term 4D recordings.

Exploring Tree Age & Diameter to Illustrate Sample Design & Inference in Observational Ecology

ERIC Educational Resources Information Center

Casady, Grant M.

2015-01-01

Undergraduate biology labs often explore the techniques of data collection but neglect the statistical framework necessary to express findings. Students can be confused about how to use their statistical knowledge to address specific biological questions. Growth in the area of observational ecology requires that students gain experience in…

The effect of extraction, storage, and analysis techniques on the measurement of airborne endotoxin from a large dairy

USDA-ARS?s Scientific Manuscript database

The objective of this study was to fill in additional knowledge gaps with respect to the extraction, storage, and analysis of airborne endotoxin, with a specific focus on samples from a dairy production facility. We utilized polycarbonate filters to collect total airborne endotoxins, sonication as ...

Statistical Modeling of the Individual: Rationale and Application of Multivariate Stationary Time Series Analysis

ERIC Educational Resources Information Center

Hamaker, Ellen L.; Dolan, Conor V.; Molenaar, Peter C. M.

2005-01-01

Results obtained with interindividual techniques in a representative sample of a population are not necessarily generalizable to the individual members of this population. In this article the specific condition is presented that must be satisfied to generalize from the interindividual level to the intraindividual level. A way to investigate…

Giardiasis: an update review on sensitivity and specificity of methods for laboratorial diagnosis.

PubMed

Soares, Renata; Tasca, Tiana

2016-10-01

Giardiasis is a major cause of diarrhoea transmitted by ingestion of contaminated water and food with cysts, and it has been spread among people with poor oral hygiene. The traditional diagnosis is performed by identifying trophozoites and cysts of Giardia duodenalis through microscopy of faecal samples. In addition to microscopy, different methods have been validated for giardiasis diagnosis which are based on immunologic and molecular analyses. The aim of this study was to conduct a review of the main methods applied in clinical laboratory for diagnosis of giardiasis, in the last 10years, regarding the specificity and sensitivity criteria. It was observed high variability in the performance of the same methodology across studies; however, several techniques have been considered better than microscopy. The later, although gold standard, presents low sensitivity in cases of low number of cysts in the sample, and the experience of the microscopist must also be considered. We conclude that microscopy should still be held and complementary technique is recommended, in order to provide a reliable diagnosis and a proper treatment of the patient. Copyright © 2016 Elsevier B.V. All rights reserved.

Expert system verification and validation study

NASA Technical Reports Server (NTRS)

French, Scott W.; Hamilton, David

1992-01-01

Five workshops on verification and validation (V&V) of expert systems (ES) where taught during this recent period of performance. Two key activities, previously performed under this contract, supported these recent workshops (1) Survey of state-of-the-practice of V&V of ES and (2) Development of workshop material and first class. The first activity involved performing an extensive survey of ES developers in order to answer several questions regarding the state-of-the-practice in V&V of ES. These questions related to the amount and type of V&V done and the successfulness of this V&V. The next key activity involved developing an intensive hands-on workshop in V&V of ES. This activity involved surveying a large number of V&V techniques, conventional as well as ES specific ones. In addition to explaining the techniques, we showed how each technique could be applied on a sample problem. References were included in the workshop material, and cross referenced to techniques, so that students would know where to go to find additional information about each technique. In addition to teaching specific techniques, we included an extensive amount of material on V&V concepts and how to develop a V&V plan for an ES project. We felt this material was necessary so that developers would be prepared to develop an orderly and structured approach to V&V. That is, they would have a process that supported the use of the specific techniques. Finally, to provide hands-on experience, we developed a set of case study exercises. These exercises were to provide an opportunity for the students to apply all the material (concepts, techniques, and planning material) to a realistic problem.

Characterization and speciation of mercury-bearing mine wastes using X-ray absorption spectroscopy

USGS Publications Warehouse

Kim, C.S.; Brown, Gordon E.; Rytuba, J.J.

2000-01-01

Mining of mercury deposits located in the California Coast Range has resulted in the release of mercury to the local environment and water supplies. The solubility, transport, and potential bioavailability of mercury are controlled by its chemical speciation, which can be directly determined for samples with total mercury concentrations greater than 100 mg kg-1 (ppm) using X-ray absorption spectroscopy (XAS). This technique has the additional benefits of being non-destructive to the sample, element-specific, relatively sensitive at low concentrations, and requiring minimal sample preparation. In this study, Hg L(III)-edge extended X-ray absorption fine structure (EXAFS) spectra were collected for several mercury mine tailings (calcines) in the California Coast Range. Total mercury concentrations of samples analyzed ranged from 230 to 1060 ppm. Speciation data (mercury phases present and relative abundances) were obtained by comparing the spectra from heterogeneous, roasted (calcined) mine tailings samples with a spectral database of mercury minerals and sorbed mercury complexes. Speciation analyses were also conducted on known mixtures of pure mercury minerals in order to assess the quantitative accuracy of the technique. While some calcine samples were found to consist exclusively of mercuric sulfide, others contain additional, more soluble mercury phases, indicating a greater potential for the release of mercury into solution. Also, a correlation was observed between samples from hot-spring mercury deposits, in which chloride levels are elevated, and the presence of mercury-chloride species as detected by the speciation analysis. The speciation results demonstrate the ability of XAS to identify multiple mercury phases in a heterogeneous sample, with a quantitative accuracy of ??25% for the mercury-containing phases considered. Use of this technique, in conjunction with standard microanalytical techniques such as X-ray diffraction and electron probe microanalysis, is beneficial in the prioritization and remediation of mercury-contaminated mine sites. (C) 2000 Elsevier Science B.V.

Effects of core retrieval, handling, and preservation on hydrate-bearing samples

NASA Astrophysics Data System (ADS)

Kneafsey, T. J.; Lu, H.; Winters, W. J.; Hunter, R. B.

2009-12-01

Recovery, preservation, storage, and transport of samples containing natural gas hydrate cause changes in the stress conditions, temperature, pressure, and hydrate saturation of samples. Sample handling at the ground surface and sample preservation, either by freezing in liquid nitrogen (LN) or repressurization using methane, provides additional time and driving forces for sample alteration. The extent to which these disturbances alter the properties of the hydrate bearing sediments (HBS) depend on specific sample handling techniques, as well as on the sample itself. HBS recovered during India’s National Gas Hydrate Program (NGHP) Expedition 01 and the 2007 BP Exploration Alaska - Department of Energy - U.S. Geological Survey (BP-DOE-USGS) Mount Elbert (ME) gas hydrate well on the Alaskan North Slope provide comparisons of sample alterations induced by multiple handling techniques. HBS samples from the NGHP and the ME projects were examined using x-ray computed tomography. Mount Elbert sand samples initially preserved in LN have non-uniform short “crack-like” low-density zones in the center that probably do not extend to the outside perimeter. Samples initially preserved by repressurization show fewer “crack-like” features and higher densities. Two samples were analyzed in detail by Lu and coworkers showing reduced hydrate saturations approaching the outer surface, while substantial hydrate remained in the central region. Non-pressure cored NGHP samples show relatively large altered regions approaching the core surface, while pressure-cored-liquid-nitrogen preserved samples have much less alteration.

Sample Preparation Report of the Fourth OPCW Confidence Building Exercise on Biomedical Sample Analysis

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

Udey, R. N.; Corzett, T. H.; Alcaraz, A.

Following the successful completion of the 3rd biomedical confidence building exercise (February 2013 – March 2013), which included the analysis of plasma and urine samples spiked at low ppb levels as part of the exercise scenario, another confidence building exercise was targeted to be conducted in 2014. In this 4th exercise, it was desired to focus specifically on the analysis of plasma samples. The scenario was designed as an investigation of an alleged use of chemical weapons where plasma samples were collected, as plasma has been reported to contain CWA adducts which remain present in the human body for severalmore » weeks (Solano et al. 2008). In the 3rd exercise most participants used the fluoride regeneration method to analyze for the presence of nerve agents in plasma samples. For the 4th biomedical exercise it was decided to evaluate the analysis of human plasma samples for the presence/absence of the VX adducts and aged adducts to blood proteins (e.g., VX-butyrylcholinesterase (BuChE) and aged BuChE adducts using a pepsin digest technique to yield nonapeptides; or equivalent). As the aging of VX-BuChE adducts is relatively slow (t1/2 = 77 hr at 37 °C [Aurbek et al. 2009]), soman (GD), which ages much more quickly (t1/2 = 9 min at 37 °C [Masson et al. 2010]), was used to simulate an aged VX sample. Additional objectives of this exercise included having laboratories assess novel OP-adducted plasma sample preparation techniques and analytical instrumentation methodologies, as well as refining/designating the reporting formats for these new techniques.« less

Contribution a la caracterisation des betons endommages par des methodes de l'acoustique non lineaire. Application a la reaction alcalis-silice

NASA Astrophysics Data System (ADS)

Kodjo, Apedovi

The aim of this thesis is to contribute to the non-destructive characterization of concrete materials damaged by alkali-silica reaction (ASR). For this purpose, some nonlinear characterization techniques have been developed, as well as a nonlinear resonance test device. In order to optimize the sensitivity of the test device, the excitation module and signal processing have been improved. The nonlinear tests were conducted on seven samples of concrete damaged by ASR, three samples of concrete damaged by heat, three concrete samples damaged mechanically and three sound concrete samples. Since, nonlinear behaviour of the material is often attribute to its micro-defects hysteretic behaviour, it was shown at first that concrete damaged by ASR exhibits an hysteresis behaviour. To conduct this study, an acoustoelastic test was set, and then nonlinear resonance test device was used for characterizing sound concrete and concrete damaged by ASR. It was shown that the nonlinear technique can be used for characterizing the material without knowing its initial state, and also for detecting early damage in the reactive material. Studies were also carried out on the effect of moisture regarding the nonlinear parameters; they allowed understanding the low values of nonlinear parameters measured on concrete samples that were kept in high moisture conditions. In order to find a specific characteristic of damage caused by ASR, the viscosity of ASR gel was used. An approach, based on static creep analysis, performed on the material, while applying the nonlinear resonance technique. The spring-damping model of Maxwell was used for the interpretation of the results. Then, the creep time was analysed on samples damaged by ASR. It appears that the ASR gel increases the creep time. Finally, the limitations of the nonlinear resonance technique for in situ application have been explained and a new applicable nonlinear technique was initiated. This technique use an external source such as a mass for making non-linearity behaviour in the material, while an ultrasound wave is investigating the medium. Keywords. Concrete, Alkali-silica reaction, Nonlinear acoustics, Nonlinearity, Hysteresis, Damage diagnostics.

Search for life on Mars: Evaluation of techniques

NASA Technical Reports Server (NTRS)

Schwartz, D. E.; Mancinelli, R. L.; White, M. R.

1995-01-01

An important question for exobiology is, did life evolve on Mars? To answer this question, experiments must be conducted on the martian surface. Given current mission constraints on mass, power, and volume, these experiments can only be performed using proposed analytical techniques such as: electron microscopy, X-ray fluorescence, X-ray diffraction, a-proton backscatter, g-ray spectrometry, differential thermal analysis, differential scanning calorimetry, pyrolysis gas chromatography, mass spectrometry, and specific element detectors. Using prepared test samples consisting of 1% organic matter (bovine serum albumin) in palagonite and a mixture of palagonite, clays, iron oxides, and evaporites, it was determined that a combination of X-ray diffraction and differential thermal analysis coupled with gas chromatography provides the best insight into the chemistry, mineralogy, and geological history of the samples.

Band Excitation for Scanning Probe Microscopy

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

Jesse, Stephen

2017-01-02

The Band Excitation (BE) technique for scanning probe microscopy uses a precisely determined waveform that contains specific frequencies to excite the cantilever or sample in an atomic force microscope to extract more information, and more reliable information from a sample. There are a myriad of details and complexities associated with implementing the BE technique. There is therefore a need to have a user friendly interface that allows typical microscopists access to this methodology. This software enables users of atomic force microscopes to easily: build complex band-excitation waveforms, set-up the microscope scanning conditions, configure the input and output electronics for generatemore » the waveform as a voltage signal and capture the response of the system, perform analysis on the captured response, and display the results of the measurement.« less

Search for life on Mars: evaluation of techniques.

PubMed

Schwartz, D E; Mancinelli, R L; White, M R

1995-03-01

An important question for exobiology is, did life evolve on Mars? To answer this question, experiments must be conducted on the martian surface. Given current mission constraints on mass, power, and volume, these experiments can only be performed using proposed analytical techniques such as: electron microscopy, X-ray fluorescence, X-ray diffraction, alpha-proton backscatter, gamma-ray spectrometry, differential thermal analysis, differential scanning calorimetry, pyrolysis gas chromatography, mass spectrometry, and specific element detectors. Using prepared test samples consisting of 1% organic matter (bovine serum albumin) in palagonite and a mixture of palagonite, clays, iron oxides, and evaporites, it was determined that a combination of X-ray diffraction and differential thermal analysis coupled with gas chromatography provides the best insight into the chemistry, mineralogy, and geological history of the samples.

Elemental and isotopic imaging of biological samples using NanoSIMS.

PubMed

Kilburn, Matt R; Clode, Peta L

2014-01-01

With its low detection limits and the ability to analyze most of the elements in the periodic table, secondary ion mass spectrometry (SIMS) represents one of the most versatile in situ analytical techniques available, and recent developments have resulted in significant advantages for the use of imaging mass spectrometry in biological and biomedical research. Increases in spatial resolution and sensitivity allow detailed interrogation of samples at relevant scales and chemical concentrations. Advances in dynamic SIMS, specifically with the advent of NanoSIMS, now allow the tracking of stable isotopes within biological systems at subcellular length scales, while static SIMS combines subcellular imaging with molecular identification. In this chapter, we present an introduction to the SIMS technique, with particular reference to NanoSIMS, and discuss its application in biological and biomedical research.

Development of an indirect immunofluorescence technique for the diagnosis of toxoplasmosis in bottlenose dolphins.

PubMed

Bernal-Guadarrama, María José; Salichs, Joan; Almunia, Javier; García-Parraga, Daniel; Fernández-Gallardo, Nuhacet; Santana-Morales, María Ángeles; Pacheco, Víctor; Afonso-Lehmann, Raquel N; Déniz, Daniel; Lorenzo-Morales, Jacob; Valladares, Basilio; Martínez-Carretero, Enrique

2014-02-01

The diagnosis of toxoplasmosis is often complicated by the lack of specific clinical symptoms or postmortem features, in humans and other animals. The only diagnostic test described so far for the serological diagnosis of Toxoplasma gondii in marine mammals is the modified agglutination test (Dubey et al., Am J Vet Res 48(8):1239-1243, 1987). The development of more sensible and specific immunological techniques requires specific antibodies, which are currently unavailable in the scientific market. Indirect immunofluorescence (IIF) is one of the most widely used methods for the diagnosis of toxoplasmosis in humans (Auer et al., Parasitol Res 12:965-970, 2000). In order to develop and apply this technique to the bottlenose dolphin (Tursiops truncatus), immunoglobulins were firstly purified using ion-exchange chromatography. The purified immunoglobulins were then injected in New Zealand rabbits in order to obtain polyclonal antibodies. These antisera were validated by the IIF technique, using as controls serum samples of dolphins infected by Toxoplasma. The results were visualized using antirabbit IgG labeled with fluorescein. This newly developed and specific serological assay was then tested with the dolphin collection of Loro Parque, Tenerife, Spain (group I), and L'Oceanogràfic of Valencia, Spain (group II). The obtained results in this study showed that none of the dolphins from group 1 were infected by T. gondii and two animals were positive in group 2. Furthermore, we conclude that this study has produced antibodies with high specificity against dolphin immunoglobulins and an IIF method which may be used as immunological diagnostic tools, especially for the serological diagnosis of toxoplasmosis.

Evaluation of a New Immunochromatography Technology Test (LDBio Diagnostics) To Detect Toxoplasma IgG and IgM: Comparison with the Routine Architect Technique

PubMed Central

Flori, Pierre; Delaunay, Edouard; Guillerme, Cécile; Charaoui, Sana; Raberin, Hélène; Hafid, Jamal; L'Ollivier, Coralie

2017-01-01

ABSTRACT A study comparing the ICT (immunochromatography technology) Toxoplasma IgG and IgM rapid diagnostic test (LDBio Diagnostics, France) with a fully automated system, Architect, was performed on samples from university hospitals of Marseille and Saint-Etienne. A total of 767 prospective sera and 235 selected sera were collected. The panels were selected to test various IgG and IgM parameters. The reference technique, Toxoplasma IgGII Western blot analysis (LDBio Diagnostics), was used to confirm the IgG results, and commercial kits Platelia Toxo IgM (Bio-Rad) and Toxo-ISAgA (bioMérieux) were used in Saint-Etienne and Marseille, respectively, as the IgM reference techniques. Sensitivity and specificity of the ICT and the Architect IgG assays were compared using a prospective panel. Sensitivity was 100% for the ICT test and 92.1% for Architect (cutoff at 1.6 IU/ml). The low-IgG-titer serum results confirmed that ICT sensitivity was superior to that of Architect. Specificity was 98.7% (ICT) and 99.8% (Architect IgG). The ICT test is also useful for detecting IgM without IgG and is both sensitive (100%) and specific (100%), as it can distinguish nonspecific IgM from specific Toxoplasma IgM. In comparison, IgM sensitivity and specificity on Architect are 96.1% and 99.6%, respectively (cutoff at 0.5 arbitrary units [AU]/ml). To conclude, this new test overcomes the limitations of automated screening techniques, which are not sensitive enough for IgG and lack specificity for IgM (rare IgM false-positive cases). PMID:28954897

Identification criteria of the rare multi-flagellate Lophomonas blattarum: comparison of different staining techniques.

PubMed

Alam-Eldin, Yosra Hussein; Abdulaziz, Amany Mamdouh

2015-09-01

Bronchopulmonary lophomoniasis (BPL) is an emerging disease of potential importance. BPL is presented by non-specific clinical picture and is usually accompanied by immunosuppression. Culture of Lophomonas blattarum is difficult and its molecular diagnosis has not yet been developed. Therefore, microscopic examination of respiratory samples, e.g., bronchoalveolar lavage (BAL) or sputum, is the mainstay of BPL diagnosis. Creola bodies and ciliocytophthoria are two forms of bronchial cells which occur in chest diseases with non-specific clinical picture like that of BPL. Both forms could be misrecognized as multi-flagellates because of their motile cilia in the wet mounts and due to shape variability of L. blattarum in stained smears. The aim of the study is to compare different staining techniques for visualizing L. blattarum to improve the recognition and diagnosis of BPL, to distinguish respiratory epithelial cells from L. blattarum and to decide which stain is recommended in suspected cases of BPL. BAL samples from patients which contain L. blattarum, creola bodies, and ciliocytophthoria were collected then wet mounts were examined. The BAL samples were also stained by Papanicolaou (PAP), Giemsa, hematoxylin and eosin (H & E), trichrome, Gram, and Diff-Quik (DQ) stains. The different staining techniques were compared regarding the stain quality. In wet mounts, the ciliary movement was coordinate and synchronous while the flagellar movement was wavy and leaded to active swimming of L. blattarum. In stained slides, bronchial cells were characterized by the presence of basal nucleus and the terminal bar from which the cilia arise. Trichrome was the best stain in demonstration of cellular details of L. blattarum. H & E, PAP, and Giemsa stains showed good quality of stains. Gram and DQ stains showed only pale hues of L. blattarum. We recommended adding Wheatley's trichrome staining to the differential diagnosis workup of cases of non-specific chest infections, especially when BPL is suspected, to avoid overdiagnosis or underdiagnosis of it.

System health monitoring using multiple-model adaptive estimation techniques

NASA Astrophysics Data System (ADS)

Sifford, Stanley Ryan

Monitoring system health for fault detection and diagnosis by tracking system parameters concurrently with state estimates is approached using a new multiple-model adaptive estimation (MMAE) method. This novel method is called GRid-based Adaptive Parameter Estimation (GRAPE). GRAPE expands existing MMAE methods by using new techniques to sample the parameter space. GRAPE expands on MMAE with the hypothesis that sample models can be applied and resampled without relying on a predefined set of models. GRAPE is initially implemented in a linear framework using Kalman filter models. A more generalized GRAPE formulation is presented using extended Kalman filter (EKF) models to represent nonlinear systems. GRAPE can handle both time invariant and time varying systems as it is designed to track parameter changes. Two techniques are presented to generate parameter samples for the parallel filter models. The first approach is called selected grid-based stratification (SGBS). SGBS divides the parameter space into equally spaced strata. The second approach uses Latin Hypercube Sampling (LHS) to determine the parameter locations and minimize the total number of required models. LHS is particularly useful when the parameter dimensions grow. Adding more parameters does not require the model count to increase for LHS. Each resample is independent of the prior sample set other than the location of the parameter estimate. SGBS and LHS can be used for both the initial sample and subsequent resamples. Furthermore, resamples are not required to use the same technique. Both techniques are demonstrated for both linear and nonlinear frameworks. The GRAPE framework further formalizes the parameter tracking process through a general approach for nonlinear systems. These additional methods allow GRAPE to either narrow the focus to converged values within a parameter range or expand the range in the appropriate direction to track the parameters outside the current parameter range boundary. Customizable rules define the specific resample behavior when the GRAPE parameter estimates converge. Convergence itself is determined from the derivatives of the parameter estimates using a simple moving average window to filter out noise. The system can be tuned to match the desired performance goals by making adjustments to parameters such as the sample size, convergence criteria, resample criteria, initial sampling method, resampling method, confidence in prior sample covariances, sample delay, and others.

Low cost coherent demodulation for mobile satellite terminals

NASA Technical Reports Server (NTRS)

Dutta, Santanu; Henely, Steven J.

1993-01-01

This paper describes some low cost approaches to coherent BPSK demodulation for mobile satellite receivers. The specific application is an Inmarsat-C Land Mobile Earth Station (LMES), but the techniques are applicable to any PSK demodulator. The techniques discussed include combined sampling and quadrature downconversion with a single A/D and novel DSP algorithms for carrier acquisition offering both superior performance and economy of DSP resources. The DSP algorithms run at 5.7 MIPS, and the entire DSP subsystem, built with commercially available parts, costs under $60 at quantity-10,000.

Bayesian linkage and segregation analysis: factoring the problem.

PubMed

Matthysse, S

2000-01-01

Complex segregation analysis and linkage methods are mathematical techniques for the genetic dissection of complex diseases. They are used to delineate complex modes of familial transmission and to localize putative disease susceptibility loci to specific chromosomal locations. The computational problem of Bayesian linkage and segregation analysis is one of integration in high-dimensional spaces. In this paper, three available techniques for Bayesian linkage and segregation analysis are discussed: Markov Chain Monte Carlo (MCMC), importance sampling, and exact calculation. The contribution of each to the overall integration will be explicitly discussed.

Multistage, multiseasonal and multiband imagery to identify and qualify non-forest vegetation resources

NASA Technical Reports Server (NTRS)

Driscoll, R. S.; Francis, R. E.

1970-01-01

A description of space and supporting aircraft photography for the interpretation and analyses of non-forest (shrubby and herbaceous) native vegetation is presented. The research includes the development of a multiple sampling technique to assign quantitative area values of specific plant community types included within an assigned space photograph map unit. Also, investigations of aerial film type, scale, and season of photography for identification and quantity measures of shrubby and herbaceous vegetation were conducted. Some work was done to develop automated interpretation techniques with film image density measurement devices.

Electrical Resistivity Measurements: a Review

NASA Astrophysics Data System (ADS)

Singh, Yadunath

World-wide interest on the use of ceramic materials for aerospace and other advanced engineering applications, has led to the need for inspection techniques capable of detecting unusually electrical and thermal anomalies in these compounds. Modern ceramic materials offer many attractive physical, electrical and mechanical properties for a wide and rapidly growing range of industrial applications; moreover specific use may be made of their electrical resistance, chemical resistance, and thermal barrier properties. In this review, we report the development and various techniques for the resistivity measurement of solid kind of samples.

Dielectrophoretic manipulation of particles for use in microfluidic devices

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

Belgrader, P; Bettencourt, K; Hamilton, J

1999-06-23

Amplification and hybridization of DNA are commonly used techniques to verify the presence of a specific DNA sequence in a test sample. Automatic sample handling to concentrate and purify sample prior to amplification is desirable both from the cost standpoint and from the standpoint of reducing the possibility of sample contamination. This paper explores the use of the dielectrophoretic force to manipulate DNA, Bacillus globigii spores, and Erwinia herbicola bacteria to provide concentration and purification as part of the sample handling functions in biological monitoring equipment. It was found that for what would be considered a typical microfabricated structure withmore » electrode gaps at 30 {micro}m operating at 5V, that concentration of the particles is very effective.« less

Application of hanging drop technique for stem cell differentiation and cytotoxicity studies.

PubMed

Banerjee, Meenal; Bhonde, Ramesh R

2006-05-01

The aim of our study is to explore the possibility of using an ancient method of culture technique- the hanging drop technique for stem cell differentiation and cytotoxicity testing. We demonstrate here a variety of novel applications of this age old technique not only to harness the differentiation potential of stem cells into specific lineages but also for cytotoxicity studies. Here we have prepared hanging drop cultures by placing 20 microl micro-drops of nutrient media and 10% Fetal Calf Serum (FCS) containing cells of interest on the lids of 60 mm dishes. Bottom plates of the dishes were filled with sterile Phosphate Buffer Saline (PBS) to avoid desiccation of samples. Lids were then placed on the bottom plates to achieve hanging drop cultures. We utilized this technique for cultivation of ciliated epithelia to study cytotoxicity and differentiation of bone marrow stromal cells. Most importantly the modified culture technique presented here is simple, economical and cost effective in terms of the time taken and the reagents required and are amenable to goal specific modification such as cytotoxicity testing. It is advantageous over the existing system in terms of retention of viability and functionality for longer duration and for providing three dimensional growth micro-environment making it useful for organotypic cultures and in vivo simulation.

From tails to toes: developing nonlethal tissue indicators of mercury exposure in five amphibian species.

PubMed

Pfleeger, Adam Z; Eagles-Smith, Collin A; Kowalski, Brandon M; Herring, Garth; Willacker, James J; Jackson, Allyson K; Pierce, John R

2016-04-01

Exposure to environmental contaminants has been implicated as a factor in global amphibian decline. Mercury (Hg) is a particularly widespread contaminant that biomagnifies in amphibians and can cause a suite of deleterious effects. However, monitoring contaminant exposure in amphibian tissues may conflict with conservation goals if lethal take is required. Thus, there is a need to develop non-lethal tissue sampling techniques to quantify contaminant exposure in amphibians. Some minimally invasive sampling techniques, such as toe-clipping, are common in population-genetic research, but it is unclear if these methods can adequately characterize contaminant exposure. We examined the relationships between mercury (Hg) concentrations in non-lethally sampled tissues and paired whole-bodies in five amphibian species. Specifically, we examined the utility of three different tail-clip sections from four salamander species and toe-clips from one anuran species. Both tail and toe-clips accurately predicted whole-body THg concentrations, but the relationships differed among species and the specific tail-clip section or toe that was used. Tail-clips comprised of the distal 0-2 cm segment performed the best across all salamander species, explaining between 82 and 92% of the variation in paired whole-body THg concentrations. Toe-clips were less effective predictors of frog THg concentrations, but THg concentrations in outer rear toes accounted for up to 79% of the variability in frog whole-body THg concentrations. These findings suggest non-lethal sampling of tails and toes has potential applications for monitoring contaminant exposure and risk in amphibians, but care must be taken to ensure consistent collection and interpretation of samples.

Rapid detection of Pseudomonas aeruginosa biomarkers in biological fluids using surface-enhanced Raman scattering

NASA Astrophysics Data System (ADS)

Wu, Xiaomeng; Chen, Jing; Zhao, Yiping; Zughaier, Susu M.

2014-05-01

Pseudomonas aeruginosa (PA) is an opportunistic pathogen that causes major infection not only in Cystic Fibrosis patients but also in chronic obstructive pulmonary disease and in critically ill patients in intensive care units. Successful antibiotic treatment of the infection relies on accurate and rapid identification of the infectious agents. Conventional microbiological detection methods usually take more than 3 days to obtain accurate results. We have developed a rapid diagnostic technique based on surface-enhanced Raman scattering to directly identify PA from biological fluids. P. aeruginosa strains, PAO1 and PA14, are cultured in lysogeny broth, and the SERS spectra of the broth show the signature Raman peaks from pyocyanin and pyoverdine, two major biomarkers that P. aeruginosa secretes during its growth, as well as lipopolysaccharides. This provides the evidence that the presence of these biomarkers can be used to indicate P. aeruginosa infection. A total of 22 clinical exhaled breath condensates (EBC) samples were obtained from subjects with CF disease and from non-CF healthy donors. SERS spectra of these EBC samples were obtained and further analyzed by both principle component analysis and partial least square-discriminant analysis (PLS-DA). PLS-DA can discriminate the samples with P. aeruginosa infection and the ones without P. aeruginosa infection at 99.3% sensitivity and 99.6% specificity. In addition, this technique can also discriminate samples from subject with CF disease and healthy donor with 97.5% sensitivity and 100% specificity. These results demonstrate the potential of using SERS of EBC samples as a rapid diagnostic tool to detect PA infection.

Nano-LISA for in vitro diagnostic applications

NASA Astrophysics Data System (ADS)

Maswadi, Saher; Glickman, Randolph D.; Elliott, Rowe; Barsalou, Norman

2011-03-01

We previously reported the detection of bacterial antigen with immunoaffinity reactions using laser optoacoustic spectroscopy and antibody-coupled gold nanorods (Ab-NR) as a contrast agent specifically targeted to the antigen of interest. The Nano-LISA (Nanoparticle Linked Immunosorbent Assay) method has been adapted to detect three very common blood-borne viral infectious agents, i.e. human T-lymphotropic virus (HTLV), human immunodeficiency virus (HIV) and hepatitis-B (Hep-B). These agents were used in a model test panel to illustrate the performance of the Nano-LISA technique. A working laboratory prototype of a Nano-LISA microplate reader-sensor was assembled and tested against the panel containing specific antigens of each of the infectious viral agents. Optoacoustic (OA) responses generated by the samples were detected using the probe beam deflection technique, an all-optical, non-contact technique. A LabView graphical user interface was developed for control of the instrument and real-time display of the test results. The detection limit of Nano-LISA is at least 1 ng/ml of viral antigen, and can reach 10 pg/ml, depending on the binding affinity of the specific detection antibody used to synthesize the Ab-NR. The method has sufficient specificity, i.e. the detection reagents do not cross-react with noncomplementary antigens. Thus, the OA microplate reader, incorporating NanoLISA, has adequate detection sensitivity and specificity for use in clinical in vitro diagnostic testing.

An information system design for watershed-wide modeling of water loss to the atmosphere using remote sensing techniques

NASA Technical Reports Server (NTRS)

Khorram, S.

1977-01-01

Results are presented of a study intended to develop a general location-specific remote-sensing procedure for watershed-wide estimation of water loss to the atmosphere by evaporation and transpiration. The general approach involves a stepwise sequence of required information definition (input data), appropriate sample design, mathematical modeling, and evaluation of results. More specifically, the remote sensing-aided system developed to evaluate evapotranspiration employs a basic two-stage two-phase sample of three information resolution levels. Based on the discussed design, documentation, and feasibility analysis to yield timely, relatively accurate, and cost-effective evapotranspiration estimates on a watershed or subwatershed basis, work is now proceeding to implement this remote sensing-aided system.

Fluorescent quenching-based quantitative detection of specific DNA/RNA using a BODIPY® FL-labeled probe or primer

PubMed Central

Kurata, Shinya; Kanagawa, Takahiro; Yamada, Kazutaka; Torimura, Masaki; Yokomaku, Toyokazu; Kamagata, Yoichi; Kurane, Ryuichiro

2001-01-01

We have developed a simple method for the quantitative detection of specific DNA or RNA molecules based on the finding that BODIPY® FL fluorescence was quenched by its interaction with a uniquely positioned guanine. This approach makes use of an oligonucleotide probe or primer containing a BODIPY® FL-modified cytosine at its 5′-end. When such a probe was hybridized with a target DNA, its fluorescence was quenched by the guanine in the target, complementary to the modified cytosine, and the quench rate was proportional to the amount of target DNA. This widely applicable technique will be used directly with larger samples or in conjunction with the polymerase chain reaction to quantify small DNA samples. PMID:11239011

Classifier performance prediction for computer-aided diagnosis using a limited dataset.

PubMed

Sahiner, Berkman; Chan, Heang-Ping; Hadjiiski, Lubomir

2008-04-01

In a practical classifier design problem, the true population is generally unknown and the available sample is finite-sized. A common approach is to use a resampling technique to estimate the performance of the classifier that will be trained with the available sample. We conducted a Monte Carlo simulation study to compare the ability of the different resampling techniques in training the classifier and predicting its performance under the constraint of a finite-sized sample. The true population for the two classes was assumed to be multivariate normal distributions with known covariance matrices. Finite sets of sample vectors were drawn from the population. The true performance of the classifier is defined as the area under the receiver operating characteristic curve (AUC) when the classifier designed with the specific sample is applied to the true population. We investigated methods based on the Fukunaga-Hayes and the leave-one-out techniques, as well as three different types of bootstrap methods, namely, the ordinary, 0.632, and 0.632+ bootstrap. The Fisher's linear discriminant analysis was used as the classifier. The dimensionality of the feature space was varied from 3 to 15. The sample size n2 from the positive class was varied between 25 and 60, while the number of cases from the negative class was either equal to n2 or 3n2. Each experiment was performed with an independent dataset randomly drawn from the true population. Using a total of 1000 experiments for each simulation condition, we compared the bias, the variance, and the root-mean-squared error (RMSE) of the AUC estimated using the different resampling techniques relative to the true AUC (obtained from training on a finite dataset and testing on the population). Our results indicated that, under the study conditions, there can be a large difference in the RMSE obtained using different resampling methods, especially when the feature space dimensionality is relatively large and the sample size is small. Under this type of conditions, the 0.632 and 0.632+ bootstrap methods have the lowest RMSE, indicating that the difference between the estimated and the true performances obtained using the 0.632 and 0.632+ bootstrap will be statistically smaller than those obtained using the other three resampling methods. Of the three bootstrap methods, the 0.632+ bootstrap provides the lowest bias. Although this investigation is performed under some specific conditions, it reveals important trends for the problem of classifier performance prediction under the constraint of a limited dataset.

[Development of a universal primers PCR-coupled liquid bead array to detect biothreat bacteria].

PubMed

Wen, Hai-yan; Wang, Jing; Liu, Heng-chuan; Sun, Xiao-hong; Yang, Yu; Hu, Kong-xin; Shan, Lin-jun

2009-10-01

To develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria. 16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed. After PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples. The suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

Simultaneous detection of six urinary pteridines and creatinine by high-performance liquid chromatography-tandem mass spectrometry for clinical breast cancer detection.

PubMed

Burton, Casey; Shi, Honglan; Ma, Yinfa

2013-11-19

Recent preliminary studies have implicated urinary pteridines as candidate biomarkers in a growing number of malignancies including breast cancer. While the developments of capillary electrophoresis-laser induced fluorescence (CE-LIF), high performance liquid chromatography (HPLC), and liquid chromatography-mass spectroscopy (LC-MS) pteridine urinalyses among others have helped to enable these findings, limitations including poor pteridine specificity, asynchronous or nonexistent renal dilution normalization, and a lack of information regarding adduct formation in mass spectrometry techniques utilizing electrospray ionization (ESI) have prevented application of these techniques to a larger clinical setting. In this study, a simple, rapid, specific, and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method has been developed and optimized for simultaneous detection of six pteridines previously implicated in breast cancer and creatinine as a renal dilution factor in urine. In addition, this study reports cationic adduct formation of urinary pteridines under ESI-positive ionization for the first time. This newly developed technique separates and detects the following six urinary pteridines: 6-biopterin, 6-hydroxymethylpterin, d-neopterin, pterin, isoxanthopterin, and xanthopterin, as well as creatinine. The method detection limit for the pteridines is between 0.025 and 0.5 μg/L, and for creatinine, it is 0.15 μg/L. The method was also validated by spiked recoveries (81-105%), reproducibility (RSD: 1-6%), and application to 25 real urine samples from breast cancer positive and negative samples through a double-blind study. The proposed technique was finally compared directly with a previously reported CE-LIF technique, concluding that additional or alternative renal dilution factors are needed for proper investigation of urinary pteridines as breast cancer biomarkers.

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.

Polymerase spiral reaction (PSR): a novel, visual isothermal amplification method for detection of canine parvovirus 2 genomic DNA.

PubMed

Gupta, Vikas; Chakravarti, Soumendu; Chander, Vishal; Majumder, Saurabh; Bhat, Shabir Ahmad; Gupta, Vivek Kumar; Nandi, Sukdeb

2017-07-01

Canine parvovirus-2 (CPV-2), which is ubiquitously distributed worldwide, causes severe and often fatal gastroenteritis in dogs. Accurate, differential and rapid diagnosis of canine parvoviral enteritis remains a challenge for clinicians. A recently developed isothermal amplification technique, polymerase spiral reaction (PSR), was optimized for the first time for a viral pathogen with reference recombinant plasmid standards from different CPV-2 antigenic variants (CPV-2, CPV-2a, CPV-2b and CPV-2c) and subsequently validated using clinical samples. Addition of chromogenic substrate SYBR Green I after the completion of the reaction resulted in bright green fluorescence in positive samples, while negative samples and a no-template control remained orange. These results were further substantiated through visualization of a laddering pattern of the PSR-amplified product in an agarose gel in positive cases and the absence of this pattern in no-template control and negative samples. The PSR assay was found to be highly specific, as it did not react with other putative canine pathogens (canine adenovirus 1 and canine distemper virus). The sensitivity of the newly developed PSR technique was compared with that of conventional PCR, real-time PCR and LAMP, using a serial tenfold dilution of canine parvovirus DNA. The detection limit of PSR was found to be at the femtogram level, which is comparable with that of real-time PCR and LAMP, which are ten times more sensitive than conventional PCR. The assay was validated using 90 clinical samples, of which 54 were found positive, while only 45 samples were positive in conventional PCR. This novel assay, which is fully compliant with the 'ASSURED' concept for disease diagnosis, provides a simple, rapid, specific, sensitive and cost-effective method for diagnosis of canine parvoviral enteritis in veterinary clinics.

Comparison of five diagnostic tests for Giardia duodenalis in fecal samples from young dogs.

PubMed

Uehlinger, Fabienne D; Naqvi, S Ali; Greenwood, Spencer J; McClure, J Trenton; Conboy, Gary; O'Handley, Ryan; Barkema, Herman W

2017-09-15

Five diagnostic tests were compared for the diagnosis of Giardia duodenalis in fecal samples of young dogs. Fecal samples were collected from 136 healthy dogs <1year old and examined using immunofluorescence antibody microscopy (IFA) after sucrose gradient centrifugation, zinc sulfate centrifugal flotation technique (ZSCT), SNAP ® Giardia test, and ProSpecT ® Giardia EZ Microplate assay. In addition, polymerase chain reaction (PCR) of the 16S rRNA gene was performed. Kappa (κ) statistic was calculated to assess diagnostic agreement between the IFA and each test. Using the IFA as the gold standard, the relative sensitivity and specificity of each test were determined. Subsequently, a Bayesian approach was used to estimate the sensitivity and specificity of each test in comparison to the IFA results. Giardia duodenalis was detected in 41% of the samples examined by IFA. The ZSCT resulted in 37% of positive samples, with a relative sensitivity and specificity of 86 and 98%, respectively. The SNAP ® Giardia test was positive in 40% of the samples, with a relative sensitivity and specificity of 91 and 96%, respectively. The ProSpecT ® test was positive in 51% of the samples, with a relative sensitivity and specificity of 100 and 83%, respectively. The relative sensitivity and specificity for PCR were 58 and 56%, respectively, with 55% of samples being PCR-positive. While the sensitivity and specificity estimates of each test in comparison to the IFA changed when using a Bayesian approach, the conclusions remained the same. While the ProSpecT ® test was the most sensitive test in this study, it is not designed for dogs and more costly than the other tests. The SNAP ® Giardia test performed similar to the ZSCT but may be more favorable because it is fast and easy to perform. Performance of the PCR was poor and the benefit of PCR may be in determining genotypes for evaluating zoonotic transfer between dogs and humans. Copyright © 2017 Elsevier B.V. All rights reserved.

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

PubMed Central

Chen, Ping-Hung; Chen, Shun-Niang; Tseng, Sheng-Hao; Deng, Ming-Jay; Lin, Yang-Wei; Sun, Yuh-Chang

2016-01-01

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions. PMID:27584954

In-situ investigation of protein and DNA structure using UVRRS

NASA Astrophysics Data System (ADS)

Greek, L. Shane; Schulze, H. Georg; Blades, Michael W.; Haynes, Charles A.; Turner, Robin F. B.

1997-05-01

Ultraviolet resonance Raman spectroscopy (UVRRS) has the potential to become a sensitive, specific, versatile bioanalytical and biophysical technique for routine investigations of proteins, DNA, and their monomeric components, as well as a variety smaller, physiologically important aromatic molecules. The transition of UVRRS from a complex, specialized spectroscopic method to a common laboratory assay depends upon several developments, including a robust sample introduction method permitting routine, in situ analysis in standard laboratory environments. To this end, we recently reported the first fiber-optic probes suitable for deep-UV pulsed laser UVRRS. In this paper, we extend this work by demonstrating the applicability of such probes to studies of biochemical relevance, including investigations of the resonance enhancement of phosphotyrosine, thermal denaturation of RNase T1, and specific and non-specific protein binding. The advantages and disadvantages of the probes are discussed with reference to sample conditions and probe design considerations.

Preanalytics in lung cancer.

PubMed

Warth, Arne; Muley, Thomas; Meister, Michael; Weichert, Wilko

2015-01-01

Preanalytic sampling techniques and preparation of tissue specimens strongly influence analytical results in lung tissue diagnostics both on the morphological but also on the molecular level. However, in contrast to analytics where tremendous achievements in the last decade have led to a whole new portfolio of test methods, developments in preanalytics have been minimal. This is specifically unfortunate in lung cancer, where usually only small amounts of tissue are at hand and optimization in all processing steps is mandatory in order to increase the diagnostic yield. In the following, we provide a comprehensive overview on some aspects of preanalytics in lung cancer from the method of sampling over tissue processing to its impact on analytical test results. We specifically discuss the role of preanalytics in novel technologies like next-generation sequencing and in the state-of the-art cytology preparations. In addition, we point out specific problems in preanalytics which hamper further developments in the field of lung tissue diagnostics.

Accounting for receptor flexibility and enhanced sampling methods in computer-aided drug design.

PubMed

Sinko, William; Lindert, Steffen; McCammon, J Andrew

2013-01-01

Protein flexibility plays a major role in biomolecular recognition. In many cases, it is not obvious how molecular structure will change upon association with other molecules. In proteins, these changes can be major, with large deviations in overall backbone structure, or they can be more subtle as in a side-chain rotation. Either way the algorithms that predict the favorability of biomolecular association require relatively accurate predictions of the bound structure to give an accurate assessment of the energy involved in association. Here, we review a number of techniques that have been proposed to accommodate receptor flexibility in the simulation of small molecules binding to protein receptors. We investigate modifications to standard rigid receptor docking algorithms and also explore enhanced sampling techniques, and the combination of free energy calculations and enhanced sampling techniques. The understanding and allowance for receptor flexibility are helping to make computer simulations of ligand protein binding more accurate. These developments may help improve the efficiency of drug discovery and development. Efficiency will be essential as we begin to see personalized medicine tailored to individual patients, which means specific drugs are needed for each patient's genetic makeup. © 2012 John Wiley & Sons A/S.

Does Nosema ceranae Wipe Out Nosema apis in Turkey?

PubMed

Ivgin Tunca, Rahşan; Oskay, Devrim; Gosterit, Ayhan; Tekin, Olgay Kaan

2016-01-01

The aim of this study was to determine the prevalence of the Nosema ceranae and Nosema apis among apiaries using both spore counts and multiplex PCR and the replacement of N. apis by N. ceranae in some regions of Turkey. A hundred honey bee samples were collected from 99 apiaries in 11 different locations in 2011-2012 in Turkey. Nosema infection degree from collected samples was determined using light microscope and molecular detection of Nosema spp. ( N. ceranae and N. apis ) was performed using specific primers by multiplex PCR. N. ceranae was only found spores in sampling areas using molecular diagnosis. N. apis was not detected in whole sampling areas using both techniques. There are no Nosema spores detected in Konya one location using two techniques. The nucleotide sequences from amplification products of the Nosema infested honeybee samples were (98%) identical with the sequence of N. ceranae for many countries deposited in the GenBank database in this study. The present study illustrated that N. ceranae is the only spores for sampled areas in 2011-2012. The study could also indicate that N. ceranae has been replaced instead of N . apis in Turkey. In addition, the prevalence of N. ceranae and two microsporodia spores effects on honey bee colonies in Turkey were needed to determine with intensive sampling, periodically.

Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests

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

Liu, Tao; Hossain, Mahmud; Schepmoes, Athena A.

2012-08-03

Sandwich immunoassay is the standard technique used in clinical labs for quantifying protein biomarkers for disease detection, monitoring and therapeutic intervention. Albeit highly sensitive, the development of a specific immunoassay is rather time-consuming and associated with extremely high cost due to the requirement for paired immunoaffinity reagents of high specificity. Recently, mass spectrometry-based methods, specifically selected reaction monitoring mass spectrometry (SRM-MS), have been increasingly applied to measure low abundance biomarker candidates in tissue and biofluids, owing to high sensitivity and specificity, simplicity of assay configuration, and great multiplexing capability. In this study, we report for the first time the developmentmore » of immunoaffinity depletion-based workflows and SRM-MS assays that enable sensitive and accurate quantification of total and free prostate-specific antigen (PSA) in serum without the requirement for specific PSA antibodies. With stable isotope dilution and external calibration, low ng/mL level detection of both total and free PSA was consistently achieved in both PSA-spiked female serum samples and actual patient serum samples. Moreover, comparison of the results obtained when SRM PSA assays and conventional immunoassays were applied to the same samples showed very good correlation (R2 values ranging from 0.90 to 0.99) in several independent clinical serum sample sets, including a set of 33 samples assayed in a blinded test. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring total and free PSA in human blood. Furthermore, simultaneous measurement of free and total PSA and many other biomarkers can be performed in a single analysis using high-resolution liquid chromatographic separation coupled with SRM-MS.« less

Ultra-sensitive high performance liquid chromatography-laser-induced fluorescence based proteomics for clinical applications.

PubMed

Patil, Ajeetkumar; Bhat, Sujatha; Pai, Keerthilatha M; Rai, Lavanya; Kartha, V B; Chidangil, Santhosh

2015-09-08

An ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique has been developed by our group at Manipal, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from volunteers (normal, and different pre-malignant/malignant conditions) were recorded using this set-up. The protein profiles were analyzed using principal component analysis (PCA) to achieve objective detection and classification of malignant, premalignant and healthy conditions with high sensitivity and specificity. The HPLC-LIF protein profiling combined with PCA, as a routine method for screening, diagnosis, and staging of cervical cancer and oral cancer, is discussed in this paper. In recent years, proteomics techniques have advanced tremendously in life sciences and medical sciences for the detection and identification of proteins in body fluids, tissue homogenates and cellular samples to understand biochemical mechanisms leading to different diseases. Some of the methods include techniques like high performance liquid chromatography, 2D-gel electrophoresis, MALDI-TOF-MS, SELDI-TOF-MS, CE-MS and LC-MS techniques. We have developed an ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from healthy and volunteers with different malignant conditions were recorded by using this set-up. The protein profile data were analyzed using principal component analysis (PCA) for objective classification and detection of malignant, premalignant and healthy conditions. The method is extremely sensitive to detect proteins with limit of detection of the order of femto-moles. The HPLC-LIF combined with PCA as a potential proteomic method for the diagnosis of oral cancer and cervical cancer has been discussed in this paper. This article is part of a Special Issue entitled: Proteomics in India. Copyright © 2015 Elsevier B.V. All rights reserved.

Molecular diagnosis of strongyloidiasis in a population of an endemic area through nested-PCR.

PubMed

Sharifdini, Meysam; Keyhani, Amir; Eshraghian, Mohammad Reza; Beigom Kia, Eshrat

2018-01-01

This study is aimed to diagnose and analyze strongyloidiasis in a population of an endemic area of Iran using nested-PCR, coupled with parasitological methods. Screening of strongyloidiasis infected people using reliable diagnostic techniques are essential to decrease the mortality and morbidity associated with this infection. Molecular methods have been proved to be highly sensitive and specific for detection of Strongyloides stercoralis in stool samples. A total of 155 fresh single stool samples were randomly collected from residents of north and northwest of Khouzestan Province, Iran. All samples were examined by parasitological methods including formalin-ether concentration and nutrient agar plate culture, and molecular method of nested-PCR. Infections with S. stercoralis were analyzed according to demographic criteria. Based on the results of nested-PCR method 15 cases (9.7%) were strongyloidiasis positive. Nested-PCR was more sensitive than parasitological techniques on single stool sampling. Elderly was the most important population index for higher infectivity with S. stercoralis . In endemic areas of S. stercoralis , old age should be considered as one of the most important risk factors of infection, especially among the immunosuppressed individuals.

[Features of skin graft in pediatric plastic surgery].

PubMed

Depoortère, C; François, C; Belkhou, A; Duquennoy-Martinot, V; Guerreschi, P

2016-10-01

Skin graft is a skin tissue fragment transferred from a donor site to a receiving site with a spontaneous revascularization. Basic process of plastic surgery, skin graft known in children, specific, warnings and refinements. It finds its indication in many pediatric cases: integumental diseases (neavus, hamartoma), acute burns and scars, traumatic loss of substance or surgically induced, congenital malformations of the hands and feet, etc. Specific skin graft techniques in children are developed: donor sites, sampling technique and procedure, early postoperative care. Especially in children, the scalp is a perfect site for split skin graft and technique is actively developed. Refinements and special cases are discussed: use of dermal matrices, allografts, xenografts, negative pressure therapy, prior skin expansion of the donor site. Results of skin graft in children are exposed: taking of graft, growth and shrinkage, pigmentation. Skin graft sometimes allows to stay the complex movement and get the best final benefit, permanent or at least temporary, in a growing being. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Detection of counterfeit electronic components through ambient mass spectrometry and chemometrics.

PubMed

Pfeuffer, Kevin P; Caldwell, Jack; Shelley, Jake T; Ray, Steven J; Hieftje, Gary M

2014-09-21

In the last several years, illicit electronic components have been discovered in the inventories of several distributors and even installed in commercial and military products. Illicit or counterfeit electronic components include a broad category of devices that can range from the correct unit with a more recent date code to lower-specification or non-working systems with altered names, manufacturers and date codes. Current methodologies for identification of counterfeit electronics rely on visual microscopy by expert users and, while effective, are very time-consuming. Here, a plasma-based ambient desorption/ionization source, the flowing atmospheric pressure afterglow (FAPA) is used to generate a mass-spectral fingerprint from the surface of a variety of discrete electronic integrated circuits (ICs). Chemometric methods, specifically principal component analysis (PCA) and the bootstrapped error-adjusted single-sample technique (BEAST), are used successfully to differentiate between genuine and counterfeit ICs. In addition, chemical and physical surface-removal techniques are explored and suggest which surface-altering techniques were utilized by counterfeiters.

A New Versatile Microarray-based Method for High Throughput Screening of Carbohydrate-active Enzymes*

PubMed Central

Vidal-Melgosa, Silvia; Pedersen, Henriette L.; Schückel, Julia; Arnal, Grégory; Dumon, Claire; Amby, Daniel B.; Monrad, Rune Nygaard; Westereng, Bjørge; Willats, William G. T.

2015-01-01

Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing together with associated bioinformatics tools have identified vast numbers of putative carbohydrate-degrading and -modifying enzymes including glycoside hydrolases and lytic polysaccharide monooxygenases. However, there is a paucity of methods for rapidly screening the activities of these enzymes. By combining the multiplexing capacity of carbohydrate microarrays with the specificity of molecular probes, we have developed a sensitive, high throughput, and versatile semiquantitative enzyme screening technique that requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme mixtures, and crude culture broths against single substrates, substrate mixtures, and biomass samples. Moreover, we show that the technique can be used to analyze both endo-acting and exo-acting glycoside hydrolases, polysaccharide lyases, carbohydrate esterases, and lytic polysaccharide monooxygenases. We demonstrate the potential of the technique by identifying the substrate specificities of purified uncharacterized enzymes and by screening enzyme activities from fungal culture broths. PMID:25657012

Evaluation of modified Dennis parasitological technique for diagnosis of bovine fascioliasis.

PubMed

Correa, Stefanya; Martínez, Yudy Liceth; López, Jessika Lissethe; Velásquez, Luz Elena

2016-02-23

Bovine fascioliasis causes important economic losses, estimated at COP$ 12,483 billion per year; its prevalence is 25% in dairy cattle. Parasitological techniques are required for it diagnosis. The Dennis technique, modified in 2002, is the one used in Colombia, but its sensitivity, specificity and validity are not known.  To evaluate the validity and performance of the modified Dennis technique for diagnosis of bovine fascioliasis using as reference test the observation of parasites in the liver.  We conducted a diagnostic evaluation study. We selected a convenience sample of discarded bovines sacrificed between March and June, 2013, in Frigocolanta for the study. We collected 25 g of feces from each animal and their liver and bile ducts were examined for Fasciola hepatica. The sensitivity, specificity, predictive positive value, predictive negative value, and validity index were calculated with 95% confidence intervals. The post-mortem evaluation was used as the gold standard.  We analyzed 180 bovines. The sensitivity and specificity of the modified Dennis technique were 73.2% (95% CI=58.4% - 87.9%) and 84.2% (95% CI= 77.7% - 90.6%), respectively. The positive predictive value was 57.7% (95% CI= 43.3% - 72.1%) and the negative one 91.4% (95% CI= 86.2% - 96.6%). The prevalence of bovine fascioliasis was 22.8% (95% CI= 16.4% - 29.2%).  The validity and the performance of the modified Dennis technique were higher than those of the traditional one, which makes it a good screening test for diagnosing fascioliasis for population and prevalence studies and during animal health campaigns.

Field Evaluation of a Coproantigen Detection Test for Fascioliasis Diagnosis and Surveillance in Human Hyperendemic Areas of Andean Countries

PubMed Central

Valero, María Adela; Periago, María Victoria; Pérez-Crespo, Ignacio; Angles, René; Villegas, Fidel; Aguirre, Carlos; Strauss, Wilma; Espinoza, José R.; Herrera, Patricia; Terashima, Angelica; Tamayo, Hugo; Engels, Dirk; Gabrielli, Albis Francesco; Mas-Coma, Santiago

2012-01-01

Background Emergence of human fascioliasis prompted a worldwide control initiative including a pilot study in a few countries. Two hyperendemic areas were chosen: Huacullani, Northern Altiplano, Bolivia, representing the Altiplanic transmission pattern with high prevalences and intensities; Cajamarca valley, Peru, representing the valley pattern with high prevalences but low intensities. Coprological sample collection, transport and study procedures were analyzed to improve individual diagnosis and subsequent treatments and surveillance activities. Therefore, a coproantigen-detection technique (MM3-COPRO ELISA) was evaluated, using classical techniques for egg detection for comparison. Methodology and Findings A total of 436 and 362 stool samples from schoolchildren of Huacullani and Cajamarca, respectively, were used. Positive samples from Huacullani were 24.77% using the MM3-COPRO technique, and 21.56% using Kato-Katz. Positive samples from Cajamarca were 11.05% using MM3-COPRO, and 5.24% using rapid sedimentation and Kato-Katz. In Huacullani, using Kato-Katz as gold standard, sensitivity and specificity were 94.68% and 98.48%, respectively, and using Kato-Katz and COPRO-ELISA test together, they were 95.68% and 100%. In Cajamarca, using rapid sedimentation and Kato-Katz together, results were 94.73% and 93.58%, and using rapid sedimentation, Kato-Katz and copro-ELISA together, they were 97.56% and 100%, respectively. There was no correlation between coproantigen detection by optical density (OD) and infection intensity by eggs per gram of feces (epg) in Cajamarca low burden cases (<400 epg), nor in Huacullani high burden cases (≥400 epg), although there was in Huacullani low burden cases (<400 epg). Six cases of egg emission appeared negative by MM3-COPRO, including one with a high egg count (1248 epg). Conclusions The coproantigen-detection test allows for high sensitivity and specificity, fast large mass screening capacity, detection in the chronic phase, early detection of treatment failure or reinfection in post-treated subjects, and usefulness in surveillance programs. However, this technique falls short when evaluating the fluke burden on its own. PMID:23029575

Mars sample return: Site selection and sample acquisition study

NASA Technical Reports Server (NTRS)

Nickle, N. (Editor)

1980-01-01

Various vehicle and mission options were investigated for the continued exploration of Mars; the cost of a minimum sample return mission was estimated; options and concepts were synthesized into program possibilities; and recommendations for the next Mars mission were made to the Planetary Program office. Specific sites and all relevant spacecraft and ground-based data were studied in order to determine: (1) the adequacy of presently available data for identifying landing sities for a sample return mission that would assure the acquisition of material from the most important geologic provinces of Mars; (2) the degree of surface mobility required to assure sample acquisition for these sites; (3) techniques to be used in the selection and drilling of rock a samples; and (4) the degree of mobility required at the two Viking sites to acquire these samples.

Use of on-site high performance liquid chromatography to evaluate the magnitude and extent of organic contaminants in aquifers

USGS Publications Warehouse

Goerlitz, D.F.; Franks, B.J.

1989-01-01

Appraisal of ground water contaminated by organic substances raises problems of difficult sample collection and timely chemical analysis. High-performance liquid chromatography was evaluated for on-site determination of specific organic contaminants in ground water samples and was used at three study sites. Organic solutes were determined directly in water samples, with little or no preparation, and usually in less than an hour after collection. This information improved sampling efficiency and was useful in screening for subsequent laboratory analysis. On two occasions, on-site analysis revealed that samples were undergoing rapid change, with major solutes being upgraded and alteration products being formed. In addition to sample stability, this technique proved valuable for monitoring other sampling factors such as compositional changes with respect to pumping, filtration, and cross contamination. -Authors

Infrared Spectroscopy as a Chemical Fingerprinting Tool

NASA Technical Reports Server (NTRS)

Huff, Timothy L.

2003-01-01

Infrared (IR) spectroscopy is a powerful analytical tool in the chemical fingerprinting of materials. Any sample material that will interact with infrared light produces a spectrum and, although normally associated with organic materials, inorganic compounds may also be infrared active. The technique is rapid, reproducible and usually non-invasive to the sample. That it is non-invasive allows for additional characterization of the original material using other analytical techniques including thermal analysis and RAMAN spectroscopic techniques. With the appropriate accessories, the technique can be used to examine samples in liquid, solid or gas phase. Both aqueous and non-aqueous free-flowing solutions can be analyzed, as can viscous liquids such as heavy oils and greases. Solid samples of varying sizes and shapes may also be examined and with the addition of microscopic IR (microspectroscopy) capabilities, minute materials such as single fibers and threads may be analyzed. With the addition of appropriate software, microspectroscopy can be used for automated discrete point or compositional surface area mapping, with the latter providing a means to record changes in the chemical composition of a material surface over a defined area. Due to the ability to characterize gaseous samples, IR spectroscopy can also be coupled with thermal processes such as thermogravimetric (TG) analyses to provide both thermal and chemical data in a single run. In this configuration, solids (or liquids) heated in a TG analyzer undergo decomposition, with the evolving gases directed into the IR spectrometer. Thus, information is provided on the thermal properties of a material and the order in which its chemical constituents are broken down during incremental heating. Specific examples of these varied applications will be cited, with data interpretation and method limitations further discussed.

Prediction of Cortisol and Progesterone Concentrations in Cow Hair Using Near-Infrared Reflectance Spectroscopy (NIRS).

PubMed

Tallo-Parra, Oriol; Albanell, Elena; Carbajal, Annais; Monclús, Laura; Manteca, Xavier; Lopez-Bejar, Manel

2017-08-01

Concentrations of different steroid hormones have been used in cows as a measure of adrenal or gonadal activity and, thus, as indicators of stress or reproductive state. Detecting cortisol and progesterone in cow hair provides a long-term integrative value of retrospective adrenal or gonadal/placental activity, respectively. Current techniques for steroid detection require a hormone-extraction procedure that involves time, several types of equipment, management of reagents, and some assay procedures (which can also be time-consuming and can destroy the samples). In contrast, near-infrared reflectance spectroscopy (NIRS) is a multi-component predictor technique, characterized as rapid, nondestructive for the sample, and reagent-free. However, as a predictor technique, NIRS needs to be calibrated and validated for each matrix, hormone, and species. The main objective of this study was to evaluate the predictive value of the NIRS technique for hair cortisol and progesterone quantification in cows by using specific enzyme immunoassay as a reference method. Hair samples from 52 adult Friesian lactating cows from a commercial dairy farm were used. Reflectance spectra of hair samples were determined with a NIR reflectance spectrophotometer before and after trimming them. Although similar results were obtained, a slightly better relationship between the reference data and NIRS predicted values was found using trimmed samples. Near infrared reflectance spectroscopy demonstrated its ability to predict cortisol and progesterone concentrations with certain accuracy (R 2  = 0.90 for cortisol and R 2  = 0.87 for progesterone). Although NIRS is far from being a complete alternative to current methodologies, the proposed equations can offer screening capability. Considering the advantages of both fields, our results open the possibility for future work on the combination of hair steroid measurement and NIRS methodology.

Histopathological and immunohistochemical approaches for the diagnosis of Pasteurellosis in swine population of Punjab.

PubMed

Bhat, Payal; Singh, Nittin Dev; Leishangthem, Geeta Devi; Kaur, Amninder; Mahajan, Vishal; Banga, Harmanjit Singh; Brar, Rajinder Singh

2016-09-01

Infectious porcine bronchopneumonia, caused by Pasteurella multocida , is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques. A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study. The collected samples were then processed for histopathological and IHC studies. Out of the total 71 lung samples, 26 samples were found to be suspected for Pasteurellosis as per the microscopic changes observed, and out of these 26 samples, 16 cases were confirmed to be positive for Pasteurellosis by IHC. Varied macroscopic changes noted in lungs were pneumonic patches with consolidation of many lobes, congestion, and focal hemorrhages. Main lesions associated with lymph nodes were its enlargement and hemorrhages. Histologically, the lung showed fibrinous and suppurative bronchopneumonia, multifocal suppuration, thickening of septa with fibrin combined with cellular infiltration and edema. The higher IHC expression of P. multocida was seen in the bronchial epithelium besides in alveolar and bronchial exudate. Moreover, on comparing the histopathological and IHC scores which were calculated on the basis of characteristic microscopic lesions and number of antigen positive cells, respectively, a significant positive correlation (r=0.4234) was found. It was concluded that swine population of Punjab is having P. multocida infection. The gross and histopathological lesions can be helpful in the preliminary diagnosis of Pasteurellosis but needs to be supplemented by other immunodiagnostic tests. Moreover, IHC technique proved to be a specific, reliable, precise, and rapid technique to supplement these conventional methods of diagnosis for Pasteurellosis.

An Undergraduate Experiment for the Measurement of Perfluorinated Surfactants in Fish Liver by Liquid Chromatography-Tandem Mass Spectrometry

ERIC Educational Resources Information Center

Stock, Naomi L.; Martin, Jonathan W.; Ye, Yun; Mabury, Scott A.

2007-01-01

A laboratory experiment that provides students a hands-on introduction to the specific techniques of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and electrospray ionization is presented. The students can thus practice the analytical principles of sample extraction, detection, quantification, and quality control using a fresh fish…

Trapping of Rift Valley Fever (RVF) vectors using Light Emitting Diode (LED) CDC traps in two arboviral disease hot spots in Kenya

USDA-ARS?s Scientific Manuscript database

Background: Mosquitoes’ response to artificial lights including color has been exploited in trap designs for improved sampling of mosquito vectors. Earlier studies suggest that mosquitoes are attracted to specific wavelengths of light and thus the need to refine techniques to increase mosquito captu...

Assessment of Sentinel Node Biopsies With Full-Field Optical Coherence Tomography.

PubMed

Grieve, Kate; Mouslim, Karima; Assayag, Osnath; Dalimier, Eugénie; Harms, Fabrice; Bruhat, Alexis; Boccara, Claude; Antoine, Martine

2016-04-01

Current techniques for the intraoperative analysis of sentinel lymph nodes during breast cancer surgery present drawbacks such as time and tissue consumption. Full-field optical coherence tomography is a novel noninvasive, high-resolution, fast imaging technique. This study investigated the use of full-field optical coherence tomography as an alternative technique for the intraoperative analysis of sentinel lymph nodes. Seventy-one axillary lymph nodes from 38 patients at Tenon Hospital were imaged minutes after excision with full-field optical coherence tomography in the pathology laboratory, before being handled for histological analysis. A pathologist performed a blind diagnosis (benign/malignant), based on the full-field optical coherence tomography images alone, which resulted in a sensitivity of 92% and a specificity of 83% (n = 65 samples). Regular feedback was given during the blind diagnosis, with thorough analysis of the images, such that features of normal and suspect nodes were identified in the images and compared with histology. A nonmedically trained imaging expert also performed a blind diagnosis aided by the reading criteria defined by the pathologist, which resulted in 85% sensitivity and 90% specificity (n = 71 samples). The number of false positives of the pathologist was reduced by 3 in a second blind reading a few months later. These results indicate that following adequate training, full-field optical coherence tomography can be an effective noninvasive diagnostic tool for extemporaneous sentinel node biopsy qualification. © The Author(s) 2015.

Amplification of Mitochondrial DNA for detection of Plasmodiumvivax in Balochistan.

PubMed

Shahwani, Muhammad Naeem; Nisar, Samia; Aleem, Abdul; Panezai, Marina; Afridi, Sarwat; Malik, Shaukat Iqbal

2017-05-01

To access a new step using PCR to amplify the targeted mtDNA sequence for detecting specifically Plasmodium vivax and its co-infections, false positive and false negative results with Plasmodium falciparum. In this study we have standardized a new technical approach in which the target mitochondrial DNA sequence (mtDNA) was amplified by using a PCR technique as a tool to detect Plasmodium spp. Species specific primers were designed to hybridize with cytochrome c oxidase gene of P. vivax (cox I) and P. falciparum (cox III). Two hundred blood samples were collected on the basis of clinical symptoms which were initially examined through microscopic analysis after preparing Giemsa stained thick and thin blood smears. Afterwards genomic DNA was extracted from all samples and was then subjected to PCR amplification by using species specific primers and amplified segments were sequenced for confirmation of results. One-hundred and thirty-two blood samples were detected as positive for malaria by PCR, out of which 64 were found to be positive by PCR and 53 by both microscopy and PCR for P.vivax infection. Nine samples were found to be false negative, one P.vivax mono infection was declared as co infection by PCR and 3 samples identified as having P.falciparum gametes were confirmed as P.vivax by PCR amplification. Sensitivity and specificity were found to be 85% and 92% respectively. Results obtained through PCR method were comparatively better and reliable than microscopy.

The significance of gtf genes in caries expression: a rapid identification of Streptococcus mutans from dental plaque of child patients.

PubMed

Mishra, Apurva; Pandey, Ramesh K; Manickam, Natesan

2015-01-01

Rapid phylogenetic and functional gene (gtfB) identification of S. mutans from the dental plaque derived from children. Dental plaque collected from fifteen patients of age group 7-12 underwent centrifugation followed by genomic DNA extraction for S. mutans. Genomic DNA was processed with S. mutans specific primers in suitable PCR condtions for phylogenetic and functional gene (gtfB) identification. The yield and results were confirmed by agarose gel electrophoresis. 1% agarose gel electrophoresis depicts the positive PCR amplification at 1,485 bp when compared with standard 1 kbp indicating the presence of S. mutans in the test sample. Another PCR reaction was set using gtfB primers specific for S. mutans for functional gene identification. 1.2% agarose gel electrophoresis was done and a positive amplication was observed at 192 bp when compared to 100 bp standards. With the advancement in molecular biology techniques, PCR based identification and quantification of the bacterial load can be done within hours using species-specific primers and DNA probes. Thus, this technique may reduce the laboratory time spend in conventional culture methods, reduces the possibility of colony identification errors and is more sensitive to culture techniques.

Critical points of DNA quantification by real-time PCR – effects of DNA extraction method and sample matrix on quantification of genetically modified organisms

PubMed Central

Cankar, Katarina; Štebih, Dejan; Dreo, Tanja; Žel, Jana; Gruden, Kristina

2006-01-01

Background Real-time PCR is the technique of choice for nucleic acid quantification. In the field of detection of genetically modified organisms (GMOs) quantification of biotech products may be required to fulfil legislative requirements. However, successful quantification depends crucially on the quality of the sample DNA analyzed. Methods for GMO detection are generally validated on certified reference materials that are in the form of powdered grain material, while detection in routine laboratories must be performed on a wide variety of sample matrixes. Due to food processing, the DNA in sample matrixes can be present in low amounts and also degraded. In addition, molecules of plant origin or from other sources that affect PCR amplification of samples will influence the reliability of the quantification. Further, the wide variety of sample matrixes presents a challenge for detection laboratories. The extraction method must ensure high yield and quality of the DNA obtained and must be carefully selected, since even components of DNA extraction solutions can influence PCR reactions. GMO quantification is based on a standard curve, therefore similarity of PCR efficiency for the sample and standard reference material is a prerequisite for exact quantification. Little information on the performance of real-time PCR on samples of different matrixes is available. Results Five commonly used DNA extraction techniques were compared and their suitability for quantitative analysis was assessed. The effect of sample matrix on nucleic acid quantification was assessed by comparing 4 maize and 4 soybean matrixes. In addition 205 maize and soybean samples from routine analysis were analyzed for PCR efficiency to assess variability of PCR performance within each sample matrix. Together with the amount of DNA needed for reliable quantification, PCR efficiency is the crucial parameter determining the reliability of quantitative results, therefore it was chosen as the primary criterion by which to evaluate the quality and performance on different matrixes and extraction techniques. The effect of PCR efficiency on the resulting GMO content is demonstrated. Conclusion The crucial influence of extraction technique and sample matrix properties on the results of GMO quantification is demonstrated. Appropriate extraction techniques for each matrix need to be determined to achieve accurate DNA quantification. Nevertheless, as it is shown that in the area of food and feed testing matrix with certain specificities is impossible to define strict quality controls need to be introduced to monitor PCR. The results of our study are also applicable to other fields of quantitative testing by real-time PCR. PMID:16907967

FT-IR spectroscopy characterization of schwannoma: a case study

NASA Astrophysics Data System (ADS)

Ferreira, Isabelle; Neto, Lazaro P. M.; das Chagas, Maurilio José; Carvalho, Luís. Felipe C. S.; dos Santos, Laurita; Ribas, Marcelo; Loddi, Vinicius; Martin, Airton A.

2016-03-01

Schwannoma are rare benign neural neoplasia. The clinical diagnosis could be improved if novel optical techniques are performed. Among these techniques, FT-IR is one of the currently techniques which has been applied for samples discrimination using biochemical information with minimum sample preparation. In this work, we report a case of a schwannoma in the cervical region. A histological examination described a benign process. An immunohistochemically examination demonstrated positivity to anti-S100 protein antibody, indicating a diagnosis of schwannoma. The aim of this analysis was to characterize FT-IR spectrum of the neoplastic and normal tissue in the fingerprint (1000-1800 cm-1) and high wavenumber region (2800-3600 cm-1). The IR spectra were collect from tumor tissue and normal nerve samples by a FT-IR spectrophotometer (Spotlight Perkin Elmer 400, USA) with 64 scans, and resolution of 4 cm-1. A total of twenty spectra were recorded (10 from schwannoma and 10 from nerve). Multivariate Analysis was used to classify the data. Through average and standard deviation analysis we observed that the main spectral change occurs at ≍1600 cm-1 (amide I) and ≍1400 cm-1 (amide III) in the fingerprint region, and in CH2/CH3 protein-lipids and OH-water vibrations for the high wavenumber region. In conclusion, FT-IR could be used as a technique for schwannoma analysis helping to establish specific diagnostic.

Development of non-destructive examination techniques for CFC-metal joints in annular geometry and their application to the manufacturing of plasma-facing components

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

Di Pietro, E.; Visca, E.; Orsini, A.

1995-12-31

The design of plasma-facing components for ITER, as for any of the envisaged next-step machines, relies heavily on the use of brazed junctions to couple armour materials to the heat sink and cooling tubes. Moreover, the typical number of brazed components and the envisaged effects of local overheating due to failure in a single brazed junction stress the importance of having a set of NDE techniques developed that can ensure the flawless quality of the joint. The qualification and application of two NDE techniques (ultrasonic and thermographic analysis) for inspection of CFC-to-metal joints is described with particular regard to themore » annular geometry typical of macroblock/monoblock solutions for divertor high-heat-flux components. The results of the eddy current inspection are not reported. The development has been focused specifically on the joint between carbon-fiber composite and TZM molybdenum alloy; techniques for the production of reference defect samples have been devised and a set of reference defect samples produced. The comparative results of the NDE inspections are reported and discussed, also on the basis of the destructive examination of the samples. The nature and size of relevant and detectable defects are discussed together with hints for a possible NDE strategy for divertor high-heat-flux components.« less

Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis

PubMed Central

Wu, Xiaoxin; Wang, Yeping; Weng, Tianhao; Hu, Chenyu; Wang, Frederick X.C.; Wu, Zhigang; Yu, Dongshan; Lu, Huoquan; Yao, Hangping

2017-01-01

Abstract The early secreted protein early secretory antigenic target 6(ESAT-6) and the culture filtrate protein 10 (CFP-10) are 2 antigens that are specific to Mycobacterium tuberculosis. These 2 antigens are good targets for tuberculosis (TB) detection. To rapidly diagnose TB across a variety of samples, we developed colloidal gold immunochromatographic strips (ICSs) based on ESAT-6 and CFP-10. The strips were evaluated using 233 samples, including sputum, plasma, and pleural effusion samples. The positive detection rates for ICSs for ESAT-6 and CFP-10 in sputum (culture-positive for M tuberculosis) were 100% and 91.2%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in plasma were 34.1% and 29.4%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in pleural effusion were 64.7% and 55.9%, respectively. Experimental analysis of culture supernatant showing that the ICS developed for ESAT-6 had a sensitivity of 100% and a specificity of 91.2%. While the ICS developed for CFP-10 had a sensitivity of 91.2% and a specificity of 88.2%. The validity of the test is limited by source of sample. The technique is sensitive and specific for samples in sputum and culture media but not for plasma or pleural effusion samples. Detection of M tuberculosis using ICSs is rapid, simple, and relatively effective; thus, ICSs are a potential screening tool for TB. PMID:29390519

Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis.

PubMed

Wu, Xiaoxin; Wang, Yeping; Weng, Tianhao; Hu, Chenyu; Wang, Frederick X C; Wu, Zhigang; Yu, Dongshan; Lu, Huoquan; Yao, Hangping

2017-12-01

The early secreted protein early secretory antigenic target 6(ESAT-6) and the culture filtrate protein 10 (CFP-10) are 2 antigens that are specific to Mycobacterium tuberculosis. These 2 antigens are good targets for tuberculosis (TB) detection.To rapidly diagnose TB across a variety of samples, we developed colloidal gold immunochromatographic strips (ICSs) based on ESAT-6 and CFP-10.The strips were evaluated using 233 samples, including sputum, plasma, and pleural effusion samples.The positive detection rates for ICSs for ESAT-6 and CFP-10 in sputum (culture-positive for M tuberculosis) were 100% and 91.2%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in plasma were 34.1% and 29.4%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in pleural effusion were 64.7% and 55.9%, respectively. Experimental analysis of culture supernatant showing that the ICS developed for ESAT-6 had a sensitivity of 100% and a specificity of 91.2%. While the ICS developed for CFP-10 had a sensitivity of 91.2% and a specificity of 88.2%.The validity of the test is limited by source of sample. The technique is sensitive and specific for samples in sputum and culture media but not for plasma or pleural effusion samples. Detection of M tuberculosis using ICSs is rapid, simple, and relatively effective; thus, ICSs are a potential screening tool for TB. Copyright © 2017 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved.

Individual-based analysis of hair corticosterone reveals factors influencing chronic stress in the American pika.

PubMed

Waterhouse, Matthew D; Sjodin, Bryson; Ray, Chris; Erb, Liesl; Wilkening, Jennifer; Russello, Michael A

2017-06-01

Glucocorticoids are often measured in wildlife to assess physiological responses to environmental or ecological stress. Hair, blood, saliva, or fecal samples are generally used depending on the timescale of the stress response being investigated and species-specific considerations. Here, we report the first use of hair samples to measure long-term corticosterone levels in the climate-sensitive American pika ( Ochotona princeps ). We validated an immunoassay-based measurement of corticosterone extracted from hair samples and compared corticosterone estimates obtained from plasma, hair, and fecal samples of nine pikas. To demonstrate an ecological application of this technique, we characterized physiological stress in 49 pikas sampled and released at eight sites along two elevational transects. Microclimate variation was measured at each site using both ambient and subsurface temperature sensors. We used an information theoretic approach to compare support for linear, mixed-effects models relating corticosterone estimates to microclimate, body size, and sex. Corticosterone was measured accurately in pika hair samples after correcting for the influence of sample mass on corticosterone extraction efficiency. Hair- and plasma-based estimates of corticosterone were weakly correlated. The best-supported model suggested that corticosterone was lower in larger, male pikas, and at locations with higher ambient temperatures in summer. Our results are consistent with a general negative relationship between body mass and glucocorticoid concentration observed across mammalian species, attributed to the higher mass-specific metabolic rates of smaller bodied animals. The higher corticosterone levels in female pikas likely reflected the physiological demands of reproduction, as observed in a wide array of mammalian species. Additionally, we establish the first direct physiological evidence for thermal stress in the American pika through nonlethal sampling of corticosterone. Interestingly, our data suggest evidence for cold stress likely induced during the summer molting period. This technique should provide a useful tool to researchers wishing to assess chronic stress in climate-sensitive mammals.

Characterization of anisotropic thermal conductivity of suspended nm-thick black phosphorus with frequency-resolved Raman spectroscopy

NASA Astrophysics Data System (ADS)

Wang, Tianyu; Han, Meng; Wang, Ridong; Yuan, Pengyu; Xu, Shen; Wang, Xinwei

2018-04-01

Frequency-resolved Raman spectroscopy (FR-Raman) is a new technique for nondestructive thermal characterization. Here, we apply this new technique to measure the anisotropic thermal conductivity of suspended nm-thick black phosphorus samples without the need of optical absorption and temperature coefficient. Four samples with thicknesses between 99.8 and 157.6 nm are studied. Based on steady state laser heating and Raman measurement of samples with a specifically designed thermal transport path, the thermal conductivity ratio (κZZ/κAC) is determined to be 1.86-3.06. Based on the FR-Raman measurements, the armchair thermal conductivity is measured as 14-22 W m-1 K-1, while the zigzag thermal conductivity is 40-63 W m-1 K-1. FR-Raman has great potential for studying the thermal properties of various nanomaterials. This study significantly advances our understanding of thermal transport in black phosphorus and facilitates the application of black phosphorus in novel devices.

Application of micro-X-ray fluorescence to chemical mapping of polar ice

NASA Astrophysics Data System (ADS)

Fourcade, M. C. Morel; Barnola, J. M.; Susini, J.; Baker, R.; Durand, G.; de Angelis, M.; Duval, P.

Synchrotron-based micro-X-ray fluorescence (μXRF) equipment has been used to analyze impurities in polar ice. A customized sample holder has been developed and the μXRF equipment has been adapted with a thermal control system to keep samples unaltered during analyses. Artificial ice samples prepared from ultra-pure water were analyzed to investigate possible contamination and/or experimental artefacts. Analyses of polar ice from Antarctica (Dome C and Vostok) confirm this μXRF technique is non-destructive and sensitive. Experiments can be reproduced to confirm or refine results by focusing on interesting spots such as crystal grain boundaries or specific inclusions. Integration times and resolution can be adjusted to optimize sensitivity. Investigation of unstable particles is possible due to the short analysis time. In addition to identification of elements in impurities, μXRF is able to determine their speciations. The accuracy and reliability of the results confirm the potential of this technique for research in glaciology.

TL and ESR based identification of gamma-irradiated frozen fish using different hydrolysis techniques

NASA Astrophysics Data System (ADS)

Ahn, Jae-Jun; Akram, Kashif; Shahbaz, Hafiz Muhammad; Kwon, Joong-Ho

2014-12-01

Frozen fish fillets (walleye Pollack and Japanese Spanish mackerel) were selected as samples for irradiation (0-10 kGy) detection trials using different hydrolysis methods. Photostimulated luminescence (PSL)-based screening analysis for gamma-irradiated frozen fillets showed low sensitivity due to limited silicate mineral contents on the samples. Same limitations were found in the thermoluminescence (TL) analysis on mineral samples isolated by density separation method. However, acid (HCl) and alkali (KOH) hydrolysis methods were effective in getting enough minerals to carry out TL analysis, which was reconfirmed through the normalization step by calculating the TL ratios (TL1/TL2). For improved electron spin resonance (ESR) analysis, alkali and enzyme (alcalase) hydrolysis methods were compared in separating minute-bone fractions. The enzymatic method provided more clear radiation-specific hydroxyapatite radicals than that of the alkaline method. Different hydrolysis methods could extend the application of TL and ESR techniques in identifying the irradiation history of frozen fish fillets.

High fidelity, radiation tolerant analog-to-digital converters

NASA Technical Reports Server (NTRS)

Wang, Charles Chang-I (Inventor); Linscott, Ivan Richard (Inventor); Inan, Umran S. (Inventor)

2012-01-01

Techniques for an analog-to-digital converter (ADC) using pipeline architecture includes a linearization technique for a spurious-free dynamic range (SFDR) over 80 deciBels. In some embodiments, sampling rates exceed a megahertz. According to a second approach, a switched-capacitor circuit is configured for correct operation in a high radiation environment. In one embodiment, the combination yields high fidelity ADC (>88 deciBel SFDR) while sampling at 5 megahertz sampling rates and consuming <60 milliWatts. Furthermore, even though it is manufactured in a commercial 0.25-.mu.m CMOS technology (1 .mu.m=12.sup.-6 meters), it maintains this performance in harsh radiation environments. Specifically, the stated performance is sustained through a highest tested 2 megarad(Si) total dose, and the ADC displays no latchup up to a highest tested linear energy transfer of 63 million electron Volts square centimeters per milligram at elevated temperature (131 degrees C.) and supply (2.7 Volts, versus 2.5 Volts nominal).

High-resolution light-sheet microscopy: a simulation of an optical illumination system for oil immersion

NASA Astrophysics Data System (ADS)

Lu, Xiang; Heintzmann, Rainer; Leischner, Ulrich

2015-09-01

Light sheet microscopy is a microscopy technique characterized by an illumination from the side, perpendicular to the direction of observation. While this is often used and easy to implement for imaging samples with water-immersion, the application in combination with oil-immersion is less often used and requires a specific optimization. In this paper we present our design of a light-sheet illumination optical system with a ~1μm illumination thickness, a long working distance through the immersion oil, and including a focusing system allowing for moving the focus-spot of the lightsheet laterally through the field of view. This optical design allows for the acquisition of fluorescence images in 3D with isotropic resolution of below 1 micrometer of whole-mount samples with a size of ~1mm diameter. This technique enables high-resolution insights in the 3D structure of biological samples, e.g. for research of insect anatomy or for imaging of biopsies in medical diagnostics.

Appreciating the difference between design-based and model-based sampling strategies in quantitative morphology of the nervous system.

PubMed

Geuna, S

2000-11-20

Quantitative morphology of the nervous system has undergone great developments over recent years, and several new technical procedures have been devised and applied successfully to neuromorphological research. However, a lively debate has arisen on some issues, and a great deal of confusion appears to exist that is definitely responsible for the slow spread of the new techniques among scientists. One such element of confusion is related to uncertainty about the meaning, implications, and advantages of the design-based sampling strategy that characterize the new techniques. In this article, to help remove this uncertainty, morphoquantitative methods are described and contrasted on the basis of the inferential paradigm of the sampling strategy: design-based vs model-based. Moreover, some recommendations are made to help scientists judge the appropriateness of a method used for a given study in relation to its specific goals. Finally, the use of the term stereology to label, more or less expressly, only some methods is critically discussed. Copyright 2000 Wiley-Liss, Inc.

Variable selection based cotton bollworm odor spectroscopic detection

NASA Astrophysics Data System (ADS)

Lü, Chengxu; Gai, Shasha; Luo, Min; Zhao, Bo

2016-10-01

Aiming at rapid automatic pest detection based efficient and targeting pesticide application and shooting the trouble of reflectance spectral signal covered and attenuated by the solid plant, the possibility of near infrared spectroscopy (NIRS) detection on cotton bollworm odor is studied. Three cotton bollworm odor samples and 3 blank air gas samples were prepared. Different concentrations of cotton bollworm odor were prepared by mixing the above gas samples, resulting a calibration group of 62 samples and a validation group of 31 samples. Spectral collection system includes light source, optical fiber, sample chamber, spectrometer. Spectra were pretreated by baseline correction, modeled with partial least squares (PLS), and optimized by genetic algorithm (GA) and competitive adaptive reweighted sampling (CARS). Minor counts differences are found among spectra of different cotton bollworm odor concentrations. PLS model of all the variables was built presenting RMSEV of 14 and RV2 of 0.89, its theory basis is insect volatilizes specific odor, including pheromone and allelochemics, which are used for intra-specific and inter-specific communication and could be detected by NIR spectroscopy. 28 sensitive variables are selected by GA, presenting the model performance of RMSEV of 14 and RV2 of 0.90. Comparably, 8 sensitive variables are selected by CARS, presenting the model performance of RMSEV of 13 and RV2 of 0.92. CARS model employs only 1.5% variables presenting smaller error than that of all variable. Odor gas based NIR technique shows the potential for cotton bollworm detection.

Police training in interviewing and interrogation methods: A comparison of techniques used with adult and juvenile suspects.

PubMed

Cleary, Hayley M D; Warner, Todd C

2016-06-01

Despite empirical progress in documenting and classifying various interrogation techniques, very little is known about how police are trained in interrogation methods, how frequently they use various techniques, and whether they employ techniques differentially with adult versus juvenile suspects. This study reports the nature and extent of formal (e.g., Reid Technique, PEACE, HUMINT) and informal interrogation training as well as self-reported technique usage in a diverse national sample (N = 340) of experienced American police officers. Officers were trained in a variety of different techniques ranging from comparatively benign pre-interrogation strategies (e.g., building rapport, observing body language or speech patterns) to more psychologically coercive techniques (e.g., blaming the victim, discouraging denials). Over half the sample reported being trained to use psychologically coercive techniques with both adults and juveniles. The majority (91%) receive informal, "on the job" interrogation training. Technique usage patterns indicate a spectrum of psychological intensity where information-gathering approaches were used most frequently and high-pressure tactics less frequently. Reid-trained officers (56%) were significantly more likely than officers without Reid training to use pre-interrogation and manipulation techniques. Across all analyses and techniques, usage patterns were identical for adult and juvenile suspects, suggesting that police interrogate youth in the same manner as adults. Overall, results suggest that training in specific interrogation methods is strongly associated with usage. Findings underscore the need for more law enforcement interrogation training in general, especially with juvenile suspects, and highlight the value of training as an avenue for reducing interrogation-induced miscarriages of justice. (PsycINFO Database Record (c) 2016 APA, all rights reserved).

GalWeight: A New and Effective Weighting Technique for Determining Galaxy Cluster and Group Membership

NASA Astrophysics Data System (ADS)

Abdullah, Mohamed H.; Wilson, Gillian; Klypin, Anatoly

2018-07-01

We introduce GalWeight, a new technique for assigning galaxy cluster membership. This technique is specifically designed to simultaneously maximize the number of bona fide cluster members while minimizing the number of contaminating interlopers. The GalWeight technique can be applied to both massive galaxy clusters and poor galaxy groups. Moreover, it is effective in identifying members in both the virial and infall regions with high efficiency. We apply the GalWeight technique to MDPL2 and Bolshoi N-body simulations, and find that it is >98% accurate in correctly assigning cluster membership. We show that GalWeight compares very favorably against four well-known existing cluster membership techniques (shifting gapper, den Hartog, caustic, SIM). We also apply the GalWeight technique to a sample of 12 Abell clusters (including the Coma cluster) using observations from the Sloan Digital Sky Survey. We conclude by discussing GalWeight’s potential for other astrophysical applications.

Bridging the gap between sample collection and laboratory analysis: using dried blood spots to identify human exposure to chemical agents

NASA Astrophysics Data System (ADS)

Hamelin, Elizabeth I.; Blake, Thomas A.; Perez, Jonas W.; Crow, Brian S.; Shaner, Rebecca L.; Coleman, Rebecca M.; Johnson, Rudolph C.

2016-05-01

Public health response to large scale chemical emergencies presents logistical challenges for sample collection, transport, and analysis. Diagnostic methods used to identify and determine exposure to chemical warfare agents, toxins, and poisons traditionally involve blood collection by phlebotomists, cold transport of biomedical samples, and costly sample preparation techniques. Use of dried blood spots, which consist of dried blood on an FDA-approved substrate, can increase analyte stability, decrease infection hazard for those handling samples, greatly reduce the cost of shipping/storing samples by removing the need for refrigeration and cold chain transportation, and be self-prepared by potentially exposed individuals using a simple finger prick and blood spot compatible paper. Our laboratory has developed clinical assays to detect human exposures to nerve agents through the analysis of specific protein adducts and metabolites, for which a simple extraction from a dried blood spot is sufficient for removing matrix interferents and attaining sensitivities on par with traditional sampling methods. The use of dried blood spots can bridge the gap between the laboratory and the field allowing for large scale sample collection with minimal impact on hospital resources while maintaining sensitivity, specificity, traceability, and quality requirements for both clinical and forensic applications.

Detection and molecular characterization of Vibrio parahaemolyticus isolated from seafood harvested along the southwest coast of India.

PubMed

Raghunath, Pendru; Acharya, Sadananda; Bhanumathi, Amarbahadur; Karunasagar, Iddya; Karunasagar, Indrani

2008-09-01

The levels of total and tdh(+)Vibrio parahaemolyticus were estimated in 83 seafood samples from southwest coast of India by colony hybridization. Conventional enrichment and isolation technique was also used to study the prevalence. Polymerase chain reaction (PCR) was performed on bacterial cell lyates for detection of total and pathogenic V. parahaemolyticus by amplification of specific genes. Of 83 samples tested, V. parahaemolyticus could be detected in 74 (89.2%) samples and tdh(+)V. parahaemolyticus in 5 (6.0%) samples by colony hybridization. V. parahaemolyticus was detected in 68 (81.9%) of 83 samples after 18 h of enrichment by PCR, and isolated from 63 (75.9%) of 83 samples by conventional isolation. The virulence genes tdh and trh could be detected in 8.4% and 25.3%, respectively, in the sample enrichment broths by PCR. Use of colony hybridization following enrichment to achieve sensitive detection of tdh(+)V. parahaemolyticus in seafood was evaluated using another set of 58 seafood samples. Thirty pathogenic V. parahaemolyticus strains isolated during the study were screened by PCR for genetic markers to be specific for the detection of the pandemic clone. Results of this study suggest that the GS-PCR may serve as a reliable genetic marker for the pandemic clone of V. parahaemolyticus.

Characterization of diesel fuel by chemical separation combined with capillary gas chromatography (GC) isotope ratio mass spectrometry (IRMS).

PubMed

Harvey, Scott D; Jarman, Kristin H; Moran, James J; Sorensen, Christina M; Wright, Bob W

2012-09-15

The purpose of this study was to perform a preliminary investigation of compound-specific isotope analysis (CSIA) of diesel fuels to evaluate whether the technique could distinguish diesel samples from different sources/locations. The ability to differentiate or correlate diesel samples could be valuable for discovering fuel tax evasion schemes or for environmental forensic studies. Two urea adduction-based techniques were used to isolate the n-alkanes from the fuel. Both carbon isotope ratio (δ(13)C) and hydrogen isotope ratio (δD) values for the n-alkanes were then determined by CSIA in each sample. The samples investigated had δ(13)C values that ranged from -30.1‰ to -26.8‰, whereas δD values ranged from -83‰ to -156‰. Plots of δD versus δ(13)C with sample n-alkane points connected in order of increasing carbon number gave well-separated clusters with characteristic shapes for each sample. Principal components analysis (PCA) with δ(13)C, δD, or combined δ(13)C and δD data was applied to extract the maximum information content. PCA scores plots could clearly differentiate the samples, thereby demonstrating the potential of this approach for distinguishing (e.g., fingerprinting) fuel samples using δ(13)C and δD values. Copyright © 2012 Elsevier B.V. All rights reserved.

Active Search on Carcasses versus Pitfall Traps: a Comparison of Sampling Methods.

PubMed

Zanetti, N I; Camina, R; Visciarelli, E C; Centeno, N D

2016-04-01

The study of insect succession in cadavers and the classification of arthropods have mostly been done by placing a carcass in a cage, protected from vertebrate scavengers, which is then visited periodically. An alternative is to use specific traps. Few studies on carrion ecology and forensic entomology involving the carcasses of large vertebrates have employed pitfall traps. The aims of this study were to compare both sampling methods (active search on a carcass and pitfall trapping) for each coleopteran family, and to establish whether there is a discrepancy (underestimation and/or overestimation) in the presence of each family by either method. A great discrepancy was found for almost all families with some of them being more abundant in samples obtained through active search on carcasses and others in samples from traps, whereas two families did not show any bias towards a given sampling method. The fact that families may be underestimated or overestimated by the type of sampling technique highlights the importance of combining both methods, active search on carcasses and pitfall traps, in order to obtain more complete information on decomposition, carrion habitat and cadaveric families or species. Furthermore, a hypothesis advanced on the reasons for the underestimation by either sampling method showing biases towards certain families. Information about the sampling techniques indicating which would be more appropriate to detect or find a particular family is provided.

Correlation spectrometer

DOEpatents

Sinclair, Michael B [Albuquerque, NM; Pfeifer, Kent B [Los Lunas, NM; Flemming, Jeb H [Albuquerque, NM; Jones, Gary D [Tijeras, NM; Tigges, Chris P [Albuquerque, NM

2010-04-13

A correlation spectrometer can detect a large number of gaseous compounds, or chemical species, with a species-specific mask wheel. In this mode, the spectrometer is optimized for the direct measurement of individual target compounds. Additionally, the spectrometer can measure the transmission spectrum from a given sample of gas. In this mode, infrared light is passed through a gas sample and the infrared transmission signature of the gasses present is recorded and measured using Hadamard encoding techniques. The spectrometer can detect the transmission or emission spectra in any system where multiple species are present in a generally known volume.

Protein blotting protocol for beginners.

PubMed

Petrasovits, Lars A

2014-01-01

The transfer and immobilization of biological macromolecules onto solid nitrocellulose or nylon (polyvinylidene difluoride (PVDF)) membranes subsequently followed by specific detection is referred to as blotting. DNA blots are called Southerns after the inventor of the technique, Edwin Southern. By analogy, RNA blots are referred to as northerns and protein blots as westerns (Burnette, Anal Biochem 112:195-203, 1981). With few exceptions, western blotting involves five steps, namely, sample collection, preparation, separation, immobilization, and detection. In this chapter, protocols for the entire process from sample collection to detection are described.

Spatially resolved in vivo plant metabolomics by laser ablation-based mass spectrometry imaging (MSI) techniques: LDI-MSI and LAESI

PubMed Central

Bartels, Benjamin; Svatoš, Aleš

2015-01-01

This short review aims to summarize the current developments and applications of mass spectrometry-based methods for in situ profiling and imaging of plants with minimal or no sample pre-treatment or manipulation. Infrared-laser ablation electrospray ionization and UV-laser desorption/ionization methods are reviewed. The underlying mechanisms of the ionization techniques–namely, laser ablation of biological samples and electrospray ionization–as well as variations of the LAESI ion source for specific targets of interest are described. PMID:26217345

The Effects of High Temperature on Gessoes with Different Admixtures

NASA Astrophysics Data System (ADS)

Budu, Ana-Maria; Sandu, Ion; Cristache, Raluca Anamaria

2014-11-01

This paper presents the effects of temperature on gessoes that have different substances added, usually used in painting or restoration to enhance the flexibility of the ground layer or to create a suitable gesso for the specific painting technique. Five samples of gesso were made and applied on Balsa wood (a dry, stable wood that is used in restoration for completing the missing elements of the panel). After the thermal treatment, the samples were analyzed optical, by microscopy and colorimetry. The results showed small differences in colour, but no cracks of the gessoes

Single Wall Nanotube Type-Specific Functionalization and Separation

NASA Technical Reports Server (NTRS)

Boul, Peter; Nikolaev, Pavel; Sosa, Edward; Arepalli, Sivaram; Yowell, Leonard

2008-01-01

Metallic single-wall carbon nanotubes were selectively solubilized in THF and separated from semiconducting nanotubes. Once separated, the functionalized metallic tubes were de-functionalized to restore their metallic band structure. Absorption and Raman spectroscopy of the enriched samples support conclusions of the enrichment of nanotube samples by metallic type. A scalable method for enriching nanotube conductive type has been developed. Raman and UV-Vis data indicate SWCNT reaction with dodecylbenzenediazonium results in metallic enrichment. It is expected that further refinement of this techniques will lead to more dramatic separations of types and diameters.

The application of compressed sensing to long-term acoustic emission-based structural health monitoring

NASA Astrophysics Data System (ADS)

Cattaneo, Alessandro; Park, Gyuhae; Farrar, Charles; Mascareñas, David

2012-04-01

The acoustic emission (AE) phenomena generated by a rapid release in the internal stress of a material represent a promising technique for structural health monitoring (SHM) applications. AE events typically result in a discrete number of short-time, transient signals. The challenge associated with capturing these events using classical techniques is that very high sampling rates must be used over extended periods of time. The result is that a very large amount of data is collected to capture a phenomenon that rarely occurs. Furthermore, the high energy consumption associated with the required high sampling rates makes the implementation of high-endurance, low-power, embedded AE sensor nodes difficult to achieve. The relatively rare occurrence of AE events over long time scales implies that these measurements are inherently sparse in the spike domain. The sparse nature of AE measurements makes them an attractive candidate for the application of compressed sampling techniques. Collecting compressed measurements of sparse AE signals will relax the requirements on the sampling rate and memory demands. The focus of this work is to investigate the suitability of compressed sensing techniques for AE-based SHM. The work explores estimating AE signal statistics in the compressed domain for low-power classification applications. In the event compressed classification finds an event of interest, ι1 norm minimization will be used to reconstruct the measurement for further analysis. The impact of structured noise on compressive measurements is specifically addressed. The suitability of a particular algorithm, called Justice Pursuit, to increase robustness to a small amount of arbitrary measurement corruption is investigated.

Sample presentation, sources of error and future perspectives on the application of vibrational spectroscopy in the wine industry.

PubMed

Cozzolino, Daniel

2015-03-30

Vibrational spectroscopy encompasses a number of techniques and methods including ultra-violet, visible, Fourier transform infrared or mid infrared, near infrared and Raman spectroscopy. The use and application of spectroscopy generates spectra containing hundreds of variables (absorbances at each wavenumbers or wavelengths), resulting in the production of large data sets representing the chemical and biochemical wine fingerprint. Multivariate data analysis techniques are then required to handle the large amount of data generated in order to interpret the spectra in a meaningful way in order to develop a specific application. This paper focuses on the developments of sample presentation and main sources of error when vibrational spectroscopy methods are applied in wine analysis. Recent and novel applications will be discussed as examples of these developments. © 2014 Society of Chemical Industry.

Label-free imaging of fatty acid content within yeast samples

NASA Astrophysics Data System (ADS)

Garrett, N.; Moger, J.

2013-02-01

Fungi have been found to be an underlying cause of 70% of all plant and animal extinctions caused by infectious diseases. Fungal infections are a growing problem affecting global health, food production and ecosystems. Lipid metabolism is a promising target for antifungal drugs and since effective treatment of fungal infections requires a better understanding of the effects of antifungal agents at the cellular level, new techniques are needed to investigate this problem. Recent advances in nonlinear microscopy allow chemically-specific contrast to be obtained non-invasively from intrinsic chemical bonds within live samples using advanced spectroscopy techniques probing Raman-active resonances. We present preliminary data using Stimulated Raman Scattering (SRS) microscopy as a means to visualise lipid droplets within individual living fungi by probing Raman resonances of the CH stretching region between 2825cm-1 and 3030cm-1.

Impact of changing from staining to culture techniques on detection rates of Campylobacter spp. in routine stool samples in Chile.

PubMed

Porte, Lorena; Varela, Carmen; Haecker, Thomas; Morales, Sara; Weitzel, Thomas

2016-05-13

Campylobacter is a leading cause of bacterial gastroenteritis, but sensitive diagnostic methods such as culture are expensive and often not available in resource limited settings. Therefore, direct staining techniques have been developed as a practical and economical alternative. We analyzed the impact of replacing Campylobacter staining with culture for routine stool examinations in a private hospital in Chile. From January to April 2014, a total of 750 consecutive stool samples were examined in parallel by Hucker stain and Campylobacter culture. Isolation rates of Campylobacter were determined and the performance of staining was evaluated against culture as the gold standard. Besides, isolation rates of Campylobacter and other enteric pathogens were compared to those of past years. Campylobacter was isolated by culture in 46 of 750 (6.1 %) stool samples. Direct staining only identified three samples as Campylobacter positive and reached sensitivity and specificity values of 6.5 and 100 %, respectively. In comparison to staining-based detection rates of previous years, we observed a significant increase of Campylobacter cases in our patients. Direct staining technique for Campylobacter had a very low sensitivity compared to culture. Staining methods might lead to a high rate of false negative results and an underestimation of the importance of campylobacteriosis. With the inclusion of Campylobacter culture, this pathogen became a leading cause of intestinal infection in our patient population.

Helminth Infections by Coprological Examination in Sheep-Dogs and Their Zoonotic Importance.

PubMed

Öge, Hatice; Öge, Semih; Özbakış, Gökben; Gürcan, I Safa

2017-03-01

This study was conducted to determine the prevalence of gastrointestinal helminths and diagnose the species of important zoonotic helminths in sheep dogs. Firstly, fecal samples were macroscopically examined; subsequently, formalin-ethyl acetate sedimentation and ZnSO4 centrifugal floatation techniques were applied for the examination of helminth eggs. PCR technique was utilized to determine the species of E. granulosus and T. canis in dogs found positive for Taenia spp. and Toxocara spp. Helminth infection was detected in 35.26% of sheep dogs. Taenia spp. was the most common helminth (12.05%), followed by Toxocara spp. (9.38%), Toxascaris leonina (6.25%), and Trichuris spp. (4.2%). The positive results in the E. granulosus and T. canis-specific PCR-based molecular tests were obtained in 14 of the Taenia egg-positive samples and in 5 of the Toxocara egg-positive samples from dogs. This study has suggested that coprophagy and feed raw offal and meat to dogs may be responsible for finding atypical helminth eggs in fecal samples from dogs in the absence of an actual infection. To make the diagnosis of their owned parasites of dogs, E. granulosus and T. canis which have zoonotic importance, feces must be examined by both conventional and copro-PCR techniques. In addition to dogs' feeding habits, other related factors must be taken into account in the epidemiology of helminth infection; thus, precaution and control measures will be more reliable.

Review of in situ derivatization techniques for enhanced bioanalysis using liquid chromatography with mass spectrometry.

PubMed

Baghdady, Yehia Z; Schug, Kevin A

2016-01-01

Accurate and specific analysis of target molecules in complex biological matrices remains a significant challenge, especially when ultra-trace detection limits are required. Liquid chromatography with mass spectrometry is often the method of choice for bioanalysis. Conventional sample preparation and clean-up methods prior to the analysis of biological fluids such as liquid-liquid extraction, solid-phase extraction, or protein precipitation are time-consuming, tedious, and can negatively affect target recovery and detection sensitivity. An alternative or complementary strategy is the use of an off-line or on-line in situ derivatization technique. In situ derivatization can be incorporated to directly derivatize target analytes in their native biological matrices, without any prior sample clean-up methods, to substitute or even enhance the extraction and preconcentration efficiency of these traditional sample preparation methods. Designed appropriately, it can reduce the number of sample preparation steps necessary prior to analysis. Moreover, in situ derivatization can be used to enhance the performance of the developed liquid chromatography with mass spectrometry-based bioanalysis methods regarding stability, chromatographic separation, selectivity, and ionization efficiency. This review presents an overview of the commonly used in situ derivatization techniques coupled to liquid chromatography with mass spectrometry-based bioanalysis to guide and to stimulate future research. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Evaluation of a nested-PCR for mycobacterium tuberculosis detection in blood and urine samples.

PubMed

da Cruz, Heidi Lacerda Alves; de Albuquerque Montenegro, Rosana; de Araújo Lima, Juliana Falcão; da Rocha Poroca, Diogo; da Costa Lima, Juliana Figueirêdo; Maria Lapa Montenegro, Lílian; Crovella, Sergio; Charifker Schindler, Haiana

2011-01-01

The polymerase chain reaction (PCR) and its variations, such as the nested-PCR, have been described as promising techniques for rapid diagnosis of tuberculosis (TB). With the aim of evaluating the usefulness of a nested-PCR method on samples of blood and urine of patients suspected of tuberculosis we analyzed 192 clinical samples, using as a molecular target the insertion element IS6110 specific of M. tuberculosis genome. Nested-PCR method showed higher sensitivity in patients with extrapulmonary tuberculosis (47.8% and 52% in blood and urine) when compared to patients with the pulmonary form of the disease (sensitivity of 29% and 26.9% in blood and urine), regardless of the type of biological sample used. The nested-PCR is a rapid technique that, even if not showing a good sensitivity, should be considered as a helpful tool especially in the extrapulmonary cases or in cases where confirmatory diagnosis is quite difficult to be achieved by routine methods. The performance of PCR-based techniques should be considered and tested in future works on other types of biological specimens besides sputum, like blood and urine, readily obtainable in most cases. The improving of M. tuberculosis nested-PCR detection in TB affected patients will give the possibility of an earlier detection of bacilli thus interrupting the transmission chain of the disease.

Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples.

PubMed

Gomes, Cláudia; Martinez-Puchol, Sandra; Pons, Maria J; Bazán, Jorge; Tinco, Carmen; del Valle, Juana; Ruiz, Joaquim

2016-03-01

The lack of an effective diagnostic tool for Carrion's disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion's disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques.

A new concept of efficient therapeutic drug monitoring using the high-resolution continuum source absorption spectrometry and the surface enhanced Raman spectroscopy

NASA Astrophysics Data System (ADS)

Xing, Yanlong; Fuss, Harald; Lademann, Jürgen; Huang, Mao Dong; Becker-Ross, Helmut; Florek, Stefan; Patzelt, Alexa; Meinke, Martina C.; Jung, Sora; Esser, Norbert

2018-04-01

In this study, a new therapeutic drug monitoring approach has been tested based on the combination of CaF molecular absorption using high-resolution continuum source absorption spectrometry (HR-CSAS) and surface enhanced Raman spectroscopy (SERS). HR-CSAS with mini graphite tube was successfully tested for clinical therapeutic drug monitoring of the fluorine-containing drug capecitabine in sweat samples of cancer patients: It showed advantageous features of high selectivity (no interference from Cl), high sensitivity (characteristic mass of 0.1 ng at CaF 583.069 nm), low sample consumption (down to 30 nL) and fast measurement (no sample pretreatment and less than 1 min of responding time) in tracing the fluorine signal out of capecitabine. However, this technique has the disadvantage of the total loss of the drug's structure information after burning the sample at very high temperature. Therefore, a new concept of combining HR-CSAS with a non-destructive spectroscopic method (SERS) was proposed for the sensitive sensing and specific identification of capecitabine. We tested and succeed in obtaining the molecular characteristics of the metabolite of capecitabine (named 5-fluorouracil) by the non-destructive SERS technique. With the results shown in this work, it is demonstrated that the combined spectroscopic technique of HR-CSAS and SERS will be very useful in efficient therapeutic drug monitoring in the future.

Spectroscopic properties for identifying sapphire samples from Ban Bo Kaew, Phrae Province, Thailand

NASA Astrophysics Data System (ADS)

Mogmued, J.; Monarumit, N.; Won-in, K.; Satitkune, S.

2017-09-01

Gemstone commercial is a high revenue for Thailand especially ruby and sapphire. Moreover, Phrae is a potential gem field located in the northern part of Thailand. The studies of spectroscopic properties are mainly to identify gemstone using advanced techniques (e.g. UV-Vis-NIR spectrophotometry, FTIR spectrometry and Raman spectroscopy). Typically, UV-Vis-NIR spectrophotometry is a technique to study the cause of color in gemstones. FTIR spectrometry is a technique to study the functional groups in gem-materials. Raman pattern can be applied to identify the mineral inclusions in gemstones. In this study, the natural sapphires from Ban Bo Kaew were divided into two groups based on colors including blue and green. The samples were analyzed by UV-Vis-NIR spectrophotometer, FTIR spectrometer and Raman spectroscope for studying spectroscopic properties. According to UV-Vis-NIR spectra, the blue sapphires show higher Fe3+/Ti4+ and Fe2+/Fe3+ absorption peaks than those of green sapphires. Otherwise, green sapphires display higher Fe3+/Fe3+ absorption peaks than blue sapphires. The FTIR spectra of both blue and green sapphire samples show the absorption peaks of -OH,-CH and CO2. The mineral inclusions such as ferrocolumbite and rutile in sapphires from this area were observed by Raman spectroscope. The spectroscopic properties of sapphire samples from Ban Bo Kaew, Phrae Province, Thailand are applied to be the specific evidence for gemstone identification.

HMM-based lexicon-driven and lexicon-free word recognition for online handwritten Indic scripts.

PubMed

Bharath, A; Madhvanath, Sriganesh

2012-04-01

Research for recognizing online handwritten words in Indic scripts is at its early stages when compared to Latin and Oriental scripts. In this paper, we address this problem specifically for two major Indic scripts--Devanagari and Tamil. In contrast to previous approaches, the techniques we propose are largely data driven and script independent. We propose two different techniques for word recognition based on Hidden Markov Models (HMM): lexicon driven and lexicon free. The lexicon-driven technique models each word in the lexicon as a sequence of symbol HMMs according to a standard symbol writing order derived from the phonetic representation. The lexicon-free technique uses a novel Bag-of-Symbols representation of the handwritten word that is independent of symbol order and allows rapid pruning of the lexicon. On handwritten Devanagari word samples featuring both standard and nonstandard symbol writing orders, a combination of lexicon-driven and lexicon-free recognizers significantly outperforms either of them used in isolation. In contrast, most Tamil word samples feature the standard symbol order, and the lexicon-driven recognizer outperforms the lexicon free one as well as their combination. The best recognition accuracies obtained for 20,000 word lexicons are 87.13 percent for Devanagari when the two recognizers are combined, and 91.8 percent for Tamil using the lexicon-driven technique.

Determination of formaldehyde by HPLC as the DNPH derivative following high-volume air sampling onto bisulfite-coated cellulose filters

NASA Astrophysics Data System (ADS)

de Andrade, Jailson B.; Tanner, Roger L.

A method is described for the specific collection of formaldehyde as hydroxymethanesulfonate on bisulfate-coated cellulose filters. Following extraction in aqueous acid and removal on unreacted bisulfite, the hydroxymethanesulfonate is decomposed by base, and HCHO is determined by DNPH (2,4-dinitrophenylhydrazine) derivatization and HPLC. Since the collection efficiency for formaldehyde is moderately high even when sampling ambient air at high-volume flow rates, a limit of detection of 0.2 ppbv is achieved with 30 min sampling times. Interference from acetaldehyde co-collected as 1-hydroxyethanesulfonate is <5% using this procedure. The technique shows promise for both short-term airborne sampling, and as a means of collecting mg-sized samples of HCHO on an inorganic matrix for carbon isotopic analyses.

Sustainable microbial water quality monitoring programme design using phage-lysis and multivariate techniques.

PubMed

Nnane, Daniel Ekane

2011-11-15

Contamination of surface waters is a pervasive threat to human health, hence, the need to better understand the sources and spatio-temporal variations of contaminants within river catchments. River catchment managers are required to sustainably monitor and manage the quality of surface waters. Catchment managers therefore need cost-effective low-cost long-term sustainable water quality monitoring and management designs to proactively protect public health and aquatic ecosystems. Multivariate and phage-lysis techniques were used to investigate spatio-temporal variations of water quality, main polluting chemophysical and microbial parameters, faecal micro-organisms sources, and to establish 'sentry' sampling sites in the Ouse River catchment, southeast England, UK. 350 river water samples were analysed for fourteen chemophysical and microbial water quality parameters in conjunction with the novel human-specific phages of Bacteroides GB-124 (Bacteroides GB-124). Annual, autumn, spring, summer, and winter principal components (PCs) explained approximately 54%, 75%, 62%, 48%, and 60%, respectively, of the total variance present in the datasets. Significant loadings of Escherichia coli, intestinal enterococci, turbidity, and human-specific Bacteroides GB-124 were observed in all datasets. Cluster analysis successfully grouped sampling sites into five clusters. Importantly, multivariate and phage-lysis techniques were useful in determining the sources and spatial extent of water contamination in the catchment. Though human faecal contamination was significant during dry periods, the main source of contamination was non-human. Bacteroides GB-124 could potentially be used for catchment routine microbial water quality monitoring. For a cost-effective low-cost long-term sustainable water quality monitoring design, E. coli or intestinal enterococci, turbidity, and Bacteroides GB-124 should be monitored all-year round in this river catchment. Copyright © 2011 Elsevier B.V. All rights reserved.

The efficacy of Sustained Natural Apophyseal Glides with and without Isometric Exercise Training in Non-specific Neck Pain.

PubMed

Ali, Abid; Shakil-Ur-Rehman, Syed; Sibtain, Fozia

2014-07-01

To determine the efficacy of Sustained Natural Apophyseal Glides (SNAGs) with and without Isometric Exercise Training Program (IETP) in Non-specific Neck Pain (NSNP) Methods: This randomized control trial of one year duration was conducted at out-patient department of Physiotherapy and Rehabilitation, Khyber Teaching Hospital (KTH) Peshawar, Pakistan from July 2012 to June 2013. The sample of 102 patients of NSNP were randomly selected through simple random sampling technique, and placed into two groups. The SNAGs manual physical therapy technique with IETP was applied on 51 patients in group A and SNAGs manual physical therapy techniques was applied alone on 51 patients in group B. The duration of intervention was 6 weeks, at 4 times per week. The Neck Disability Index (NDI) and Visual Analog Scale (VAS) for neck pain were assessment tools used for all patients before and after 6 weeks of physical therapy intervention. All the patients were assessed through NDI and VAS before intervention and at the completion of 6 weeks program. The data of all 102 was analyzed by SPSS-20 and statistical test was applied at 95% level of significance determine the efficacy of both the treatments interventions and compare with each other. The patients in group A, treated with SNAGs and followed by IETP for 6 weeks, demonstrated more improvement in pain and physical activity as assessed by VAS (p=0.013) and NDI (p=0.003), as compared to the patients treated with SNAGS alone, as pain and function assessed by VAS (p=0.047) and NDI (p=0.164). In group A the NDI score improved from 40 to 15 and VAS from 7 to 4, while in group B the NDI score improved from 42 to 30 and VAS from 7 to 4. Patients with non-specific neck pain treated with SNAGs manual physical therapy techniques and followed by IETP was more effective in reduction of pain and enhancement of function, as compared to those patients treated with SNAGs manual physical therapy techniques alone.

Methods development for total organic carbon accountability

NASA Technical Reports Server (NTRS)

Benson, Brian L.; Kilgore, Melvin V., Jr.

1991-01-01

This report describes the efforts completed during the contract period beginning November 1, 1990 and ending April 30, 1991. Samples of product hygiene and potable water from WRT 3A were supplied by NASA/MSFC prior to contract award on July 24, 1990. Humidity condensate samples were supplied on August 3, 1990. During the course of this contract chemical analyses were performed on these samples to qualitatively determine specific components comprising, the measured organic carbon concentration. In addition, these samples and known standard solutions were used to identify and develop methodology useful to future comprehensive characterization of similar samples. Standard analyses including pH, conductivity, and total organic carbon (TOC) were conducted. Colorimetric and enzyme linked assays for total protein, bile acid, B-hydroxybutyric acid, methylene blue active substances (MBAS), urea nitrogen, ammonia, and glucose were also performed. Gas chromatographic procedures for non-volatile fatty acids and EPA priority pollutants were also performed. Liquid chromatography was used to screen for non-volatile, water soluble compounds not amenable to GC techniques. Methods development efforts were initiated to separate and quantitate certain chemical classes not classically analyzed in water and wastewater samples. These included carbohydrates, organic acids, and amino acids. Finally, efforts were initiated to identify useful concentration techniques to enhance detection limits and recovery of non-volatile, water soluble compounds.

Comparison of a gel microcolumn assay with the conventional tube test for red blood cell alloantibody titration.

PubMed

Finck, Rachel; Lui-Deguzman, Carrie; Teng, Shih-Mao; Davis, Rebecca; Yuan, Shan

2013-04-01

Titration is a semiquantitative method used to estimate red blood cell (RBC) alloantibody reactivity. The conventional tube test (CTT) technique is the traditional method for performing titration studies. The gel microcolumn assay (GMA) is also a sensitive method to detect RBC alloantibodies. The aim of this study was to compare a GMA with the CTT technique in the performance of Rh and K alloantibody titration. Patient serum samples that contained an RBC alloantibody with a singular specificity were identified by routine blood bank workflow. Parallel titration studies were performed on these samples by both the CTT method and a GMA (ID-Micro Typing System anti-IgG gel card, Micro Typing Systems, Inc., an Ortho-Clinical Diagnostics Company). Forty-eight samples were included, including 11 anti-D, five anti-c, 13 anti-E, one anti-C, three anti-e, and 15 anti-K. Overall, the two methods generated identical results in 21 of 48 samples. For 42 samples (87.5%) the two methods generated results that were within one serial dilution, and for the remaining six samples, results were within two dilutions. GMA systems may perform comparably to the CTT in titrating alloantibodies to Rh and Kell antigens. © 2012 American Association of Blood Banks.

Enhanced Quality Control in Pharmaceutical Applications by Combining Raman Spectroscopy and Machine Learning Techniques

NASA Astrophysics Data System (ADS)

Martinez, J. C.; Guzmán-Sepúlveda, J. R.; Bolañoz Evia, G. R.; Córdova, T.; Guzmán-Cabrera, R.

2018-06-01

In this work, we applied machine learning techniques to Raman spectra for the characterization and classification of manufactured pharmaceutical products. Our measurements were taken with commercial equipment, for accurate assessment of variations with respect to one calibrated control sample. Unlike the typical use of Raman spectroscopy in pharmaceutical applications, in our approach the principal components of the Raman spectrum are used concurrently as attributes in machine learning algorithms. This permits an efficient comparison and classification of the spectra measured from the samples under study. This also allows for accurate quality control as all relevant spectral components are considered simultaneously. We demonstrate our approach with respect to the specific case of acetaminophen, which is one of the most widely used analgesics in the market. In the experiments, commercial samples from thirteen different laboratories were analyzed and compared against a control sample. The raw data were analyzed based on an arithmetic difference between the nominal active substance and the measured values in each commercial sample. The principal component analysis was applied to the data for quantitative verification (i.e., without considering the actual concentration of the active substance) of the difference in the calibrated sample. Our results show that by following this approach adulterations in pharmaceutical compositions can be clearly identified and accurately quantified.

Efficacy of a lead based paint XRF analyzer and a commercially available colorimetric lead test kit as qualitative field tools for determining presence of lead in religious powders.

PubMed

Shah, Manthan P; Shendell, Derek G; Meng, Qingyu; Ohman-Strickland, Pamela; Halperin, William

2018-04-23

The performances of a portable X-Ray Fluorescence (XRF) lead paint analyzer (RMD LPA-1, Protec Instrument Corp., Waltham, MA) and a commercially available colorimetric lead test kit (First Alert Lead Test Kit, eAccess Solutions, Inc., Palatine, IL) were evaluated for use by local or state health departments as potential cost-effective rapid analysis or "spot test" field techniques for tentative identification of lead content in sindoor powders. For both field-sampling methods, sensitivity, specificity and predictive values varied widely for samples containing <300,000 μg/g lead. For samples containing ≥300,000 μg/g lead, the aforementioned metrics were 100% (however, the CIs had a wide range). In addition, both field sampling methods showed clear, consistent positive readings only for samples containing ≥300,000 μg/g lead. Even samples with lead content as high as 5,110 μg/g were not positively identified by either field analysis technique. The results of this study suggest the XRF analyzer and colorimetric lead test kit cannot be used as a rapid field test for sindoor by health department inspectors.

Non-Destructive Study of Bulk Crystallinity and Elemental Composition of Natural Gold Single Crystal Samples by Energy-Resolved Neutron Imaging

PubMed Central

Tremsin, Anton S.; Rakovan, John; Shinohara, Takenao; Kockelmann, Winfried; Losko, Adrian S.; Vogel, Sven C.

2017-01-01

Energy-resolved neutron imaging enables non-destructive analyses of bulk structure and elemental composition, which can be resolved with high spatial resolution at bright pulsed spallation neutron sources due to recent developments and improvements of neutron counting detectors. This technique, suitable for many applications, is demonstrated here with a specific study of ~5–10 mm thick natural gold samples. Through the analysis of neutron absorption resonances the spatial distribution of palladium (with average elemental concentration of ~0.4 atom% and ~5 atom%) is mapped within the gold samples. At the same time, the analysis of coherent neutron scattering in the thermal and cold energy regimes reveals which samples have a single-crystalline bulk structure through the entire sample volume. A spatially resolved analysis is possible because neutron transmission spectra are measured simultaneously on each detector pixel in the epithermal, thermal and cold energy ranges. With a pixel size of 55 μm and a detector-area of 512 by 512 pixels, a total of 262,144 neutron transmission spectra are measured concurrently. The results of our experiments indicate that high resolution energy-resolved neutron imaging is a very attractive analytical technique in cases where other conventional non-destructive methods are ineffective due to sample opacity. PMID:28102285

Influence of disorder on the signature of the pseudogap and multigap superconducting behavior in FeSe

NASA Astrophysics Data System (ADS)

Rößler, Sahana; Huang, Chien-Lung; Jiao, Lin; Koz, Cevriye; Schwarz, Ulrich; Wirth, Steffen

2018-03-01

We investigated several FeSe single crystals grown by two different methods by utilizing experimental techniques, namely, resistivity, magnetoresistance, specific heat, scanning tunneling microscopy, and spectroscopy. The residual resistivity ratio (RRR) shows systematic differences between samples grown by chemical vapor transport and flux vapor transport, indicating variance in the amount of scattering centers. Although the superconducting transition temperature Tc is not directly related to RRR, our study evidences subtle differences in the features of an incipient ordering mode related to a depletion of density of states at the Fermi level. For instance, the onset temperature of anisotropic spin fluctuations at T*≈75 K, and the temperature of the opening up of a partial gap in the density of states at T**≈30 K, are not discernible in the samples with lower RRR. Further, we show that the functional dependence of the electronic specific heat below 2 K, which allows us to determine the nodal features as well as the small superconducting gap, differs significantly in crystals grown by these two different methods. Our investigation suggests that some of the controversies about the driving mechanism for the superconducting gap or its structure and symmetry are related to minute differences in the crystals arising due to the growth techniques used and the total amount of scattering centers present in the sample.

The Multiwavelength Survey by Yale-Chile (MUSYC): Deep Near-Infrared Imaging and the Selection of Distant Galaxies

NASA Astrophysics Data System (ADS)

Quadri, Ryan; Marchesini, Danilo; van Dokkum, Pieter; Gawiser, Eric; Franx, Marijn; Lira, Paulina; Rudnick, Gregory; Urry, C. Megan; Maza, José; Kriek, Mariska; Barrientos, L. Felipe; Blanc, Guillermo A.; Castander, Francisco J.; Christlein, Daniel; Coppi, Paolo S.; Hall, Patrick B.; Herrera, David; Infante, Leopoldo; Taylor, Edward N.; Treister, Ezequiel; Willis, Jon P.

2007-09-01

We present deep near-infrared JHK imaging of four 10' × 10' fields. The observations were carried out as part of the Multiwavelength Survey by Yale-Chile (MUSYC) with ISPI on the CTIO 4 m telescope. The typical point-source limiting depths are J ~ 22.5, H ~ 21.5, and K ~ 21 (5 σ Vega). The effective seeing in the final images is ~1.0″. We combine these data with MUSYC UBVRIz imaging to create K-selected catalogs that are unique for their uniform size, depth, filter coverage, and image quality. We investigate the rest-frame optical colors and photometric redshifts of galaxies that are selected using common color selection techniques, including distant red galaxies (DRGs), star-forming and passive BzKs, and the rest-frame UV-selected BM, BX, and Lyman break galaxies (LBGs). These techniques are effective at isolating large samples of high-redshift galaxies, but none provide complete or uniform samples across the targeted redshift ranges. The DRG and BM/BX/LBG criteria identify populations of red and blue galaxies, respectively, as they were designed to do. The star-forming BzKs have a very wide redshift distribution, extending down to z ~ 1, a wide range of colors, and may include galaxies with very low specific star formation rates. In comparison, the passive BzKs are fewer in number, have a different distribution of K magnitudes, and have a somewhat different redshift distribution. By combining either the DRG and BM/BX/LBG criteria, or the star-forming and passive BzK criteria, it appears possible to define a reasonably complete sample of galaxies to our flux limit over specific redshift ranges. However, the redshift dependence of both the completeness and sampled range of rest-frame colors poses an ultimate limit to the usefulness of these techniques.

High resolution x-ray microtomography of biological samples: Requirements and strategies for satisfying them

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

Loo, B.W. Jr.

High resolution x-ray microscopy has been made possible in recent years primarily by two new technologies: microfabricated diffractive lenses for soft x-rays with about 30-50 nm resolution, and high brightness synchrotron x-ray sources. X-ray microscopy occupies a special niche in the array of biological microscopic imaging methods. It extends the capabilities of existing techniques mainly in two areas: a previously unachievable combination of sub-visible resolution and multi-micrometer sample size, and new contrast mechanisms. Because of the soft x-ray wavelengths used in biological imaging (about 1-4 nm), XM is intermediate in resolution between visible light and electron microscopies. Similarly, the penetrationmore » depth of soft x-rays in biological materials is such that the ideal sample thickness for XM falls in the range of 0.25 - 10 {mu}m, between that of VLM and EM. XM is therefore valuable for imaging of intermediate level ultrastructure, requiring sub-visible resolutions, in intact cells and subcellular organelles, without artifacts produced by thin sectioning. Many of the contrast producing and sample preparation techniques developed for VLM and EM also work well with XM. These include, for example, molecule specific staining by antibodies with heavy metal or fluorescent labels attached, and sectioning of both frozen and plastic embedded tissue. However, there is also a contrast mechanism unique to XM that exists naturally because a number of elemental absorption edges lie in the wavelength range used. In particular, between the oxygen and carbon absorption edges (2.3 and 4.4 nm wavelength), organic molecules absorb photons much more strongly than does water, permitting element-specific imaging of cellular structure in aqueous media, with no artifically introduced contrast agents. For three-dimensional imaging applications requiring the capabilities of XM, an obvious extension of the technique would therefore be computerized x-ray microtomography (XMT).« less

Evaluation of digestion methods for analysis of trace metals in mammalian tissues and NIST 1577c.

PubMed

Binder, Grace A; Metcalf, Rainer; Atlas, Zachary; Daniel, Kenyon G

2018-02-15

Digestion techniques for ICP analysis have been poorly studied for biological samples. This report describes an optimized method for analysis of trace metals that can be used across a variety of sample types. Digestion methods were tested and optimized with the analysis of trace metals in cancerous as compared to normal tissue as the end goal. Anthropological, forensic, oncological and environmental research groups can employ this method reasonably cheaply and safely whilst still being able to compare between laboratories. We examined combined HNO 3 and H 2 O 2 digestion at 170 °C for human, porcine and bovine samples whether they are frozen, fresh or lyophilized powder. Little discrepancy is found between microwave digestion and PFA Teflon pressure vessels. The elements of interest (Cu, Zn, Fe and Ni) yielded consistently higher and more accurate values on standard reference material than samples heated to 75 °C or samples that utilized HNO 3 alone. Use of H 2 SO 4 does not improve homogeneity of the sample and lowers precision during ICP analysis. High temperature digestions (>165 °C) using a combination of HNO 3 and H 2 O 2 as outlined are proposed as a standard technique for all mammalian tissues, specifically, human tissues and yield greater than 300% higher values than samples digested at 75 °C regardless of the acid or acid combinations used. The proposed standardized technique is designed to accurately quantify potential discrepancies in metal loads between cancerous and healthy tissues and applies to numerous tissue studies requiring quick, effective and safe digestions. Copyright © 2017 Elsevier Inc. All rights reserved.

Development of a Loop Mediated Isothermal Amplification (LAMP) - Surface Enhanced Raman spectroscopy (SERS) Assay for the Detection of Salmonella Enterica Serotype Enteritidis.

PubMed

Draz, Mohamed Shehata; Lu, Xiaonan

2016-01-01

As a major foodborne pathogen, Salmonella enterica serotype Enteritidis is increasingly rising as a global health concern. Here, we developed an integrated assay that combines loop mediated isothermal amplification (LAMP) and surface enhanced Raman spectroscopy (SERS) for DNA detection of S. Enteritidis using specifically designed Raman active Au-nanoprobes. The target DNA was amplified by LAMP and then labeled with Au-nanoprobes comprised of gold nanoparticle-modified with specific cy5/DNA probes to allow the detection by SERS. The sensitivity of the developed LAMP-SERS detection assay (66 CFU/mL) was ~100-fold higher than the conventional polymerase chain reaction (PCR) method. Significantly, this technique allowed highly specific detection of the target DNA of S. Enteritidis and could differentiate it from the DNA of closely related bacterial species or non-specific contamination, making it more accurate and reliable than the standard LAMP technique. The applicability of detection of S. Enteritidis in milk samples using LAMP-SERS assay was validated as well. In sum, the developed LAMP-SERS assay is highly specific and sensitive, and has the potential to be applied for rapid detection of different foodborne pathogens and other microbial contaminants.

Biological application of laser induced breakdown spectroscopy technique for determination of trace elements in hair.

PubMed

Emara, Elshaimaa M; Imam, Hisham; Hassan, Mouyed A; Elnaby, Salah H

2013-12-15

Analysis of trace elements in mammalian hair has the potential to reveal retrospective information about an individual's nutritional status and exposure. As trace elements are incorporated into the hair during the growth process, longitudinal segments of the hair may reflect the body burden during growth. Using LIBS technique, Na, K, Ca, Mg, Si, Fe, Pb and Zn were detected in a single strand of horse hair. The results obtained through LIBS technique on hair samples were compared with the traditional technique (AAS) on digested acidified solution of the same samples. The effects of the experimental parameters on the emission lines were studied and the local thermodynamic equilibrium (LTE) in produced plasma was investigated. The transient plasma condition was verified at specific time region (1500-2000 ns) in the plasma evolution corresponding to its dynamic expanding characteristic. The relative mass concentrations of Fe and Zn were calculated by setting the concentration of C as the calibration. The information obtained from the trace elements' spectra of horse hair in this study substantiates the potential of hair as a biomarker. © 2013 Elsevier B.V. All rights reserved.

Spectral pattern recognition of controlled substances in street samples using artificial neural network system

NASA Astrophysics Data System (ADS)

Poryvkina, Larisa; Aleksejev, Valeri; Babichenko, Sergey M.; Ivkina, Tatjana

2011-04-01

The NarTest fluorescent technique is aimed at the detection of analyte of interest in street samples by recognition of its specific spectral patterns in 3-dimentional Spectral Fluorescent Signatures (SFS) measured with NTX2000 analyzer without chromatographic or other separation of controlled substances from a mixture with cutting agents. The illicit drugs have their own characteristic SFS features which can be used for detection and identification of narcotics, however typical street sample consists of a mixture with cutting agents: adulterants and diluents. Many of them interfere the spectral shape of SFS. The expert system based on Artificial Neural Networks (ANNs) has been developed and applied for such pattern recognition in SFS of street samples of illicit drugs.

No Evidence for Infection of UK Prostate Cancer Patients with XMRV, BK Virus, Trichomonas vaginalis or Human Papilloma Viruses

PubMed Central

Groom, Harriet C. T.; Warren, Anne Y.; Neal, David E.; Bishop, Kate N.

2012-01-01

The prevalence of specific infections in UK prostate cancer patients was investigated. Serum from 84 patients and 62 controls was tested for neutralisation of xenotropic murine leukaemia virus-related virus (XMRV) Envelope. No reactivity was found in the patient samples. In addition, a further 100 prostate DNA samples were tested for XMRV, BK virus, Trichomonas vaginalis and human papilloma viruses by nucleic acid detection techniques. Despite demonstrating DNA integrity and assay sensitivity, we failed to detect the presence of any of these agents in DNA samples, bar one sample that was weakly positive for HPV16. Therefore we conclude that these infections are absent in this typical cohort of men with prostate cancer. PMID:22470540

No evidence for infection of UK prostate cancer patients with XMRV, BK virus, Trichomonas vaginalis or human papilloma viruses.

PubMed

Groom, Harriet C T; Warren, Anne Y; Neal, David E; Bishop, Kate N

2012-01-01

The prevalence of specific infections in UK prostate cancer patients was investigated. Serum from 84 patients and 62 controls was tested for neutralisation of xenotropic murine leukaemia virus-related virus (XMRV) Envelope. No reactivity was found in the patient samples. In addition, a further 100 prostate DNA samples were tested for XMRV, BK virus, Trichomonas vaginalis and human papilloma viruses by nucleic acid detection techniques. Despite demonstrating DNA integrity and assay sensitivity, we failed to detect the presence of any of these agents in DNA samples, bar one sample that was weakly positive for HPV16. Therefore we conclude that these infections are absent in this typical cohort of men with prostate cancer.

Image-based ELISA on an activated polypropylene microtest plate--a spectrophotometer-free low cost assay technique.

PubMed

Parween, Shahila; Nahar, Pradip

2013-10-15

In this communication, we report ELISA technique on an activated polypropylene microtest plate (APPµTP) as an illustrative example of a low cost diagnostic assay. Activated test zone in APPµTP binds a capture biomolecule through covalent linkage thereby, eliminating non-specific binding often prevalent in absorption based techniques. Efficacy of APPµTP is demonstrated by detecting human immunoglobulin G (IgG), human immunoglobulin E (IgE) and Aspergillus fumigatus antibody in patient's sera. Detection is done by taking the image of the assay solution by a desktop scanner and analyzing the color of the image. Human IgE quantification by color saturation in the image-based assay shows excellent correlation with absorbance-based assay (Pearson correlation coefficient, r=0.992). Significance of the relationship is seen from its p value which is 4.087e-11. Performance of APPµTP is also checked with respect to microtiter plate and paper-based ELISA. APPµTP can quantify an analyte as precisely as in microtiter plate with insignificant non-specific binding, a necessary prerequisite for ELISA assay. In contrast, paper-ELISA shows high non-specific binding in control sera (false positive). Finally, we have carried out ELISA steps on APPµTP by ultrasound waves on a sonicator bath and the results show that even in 8 min, it can convincingly differentiate a test sample from a control sample. In short, spectrophotometer-free image-based miniaturized ELISA on APPµTP is precise, reliable, rapid, and sensitive and could be a good substitute for conventional immunoassay procedures widely used in clinical and research laboratories. Copyright © 2013 Elsevier B.V. All rights reserved.

[Assessment of two DNA extraction methods to amplify the pneumolysin gene (PLY) from blood culture samples of Streptococcus pneumoniae].

PubMed

Hernández, Carolina; Durán, Claudia; Ulloa, María Teresa; Prado, Valeria

2004-05-01

Streptococcus pneumoniae is a common etiologic agent of invasive respiratory infections among children under 5 years of age and older adults. Isolation rates of S. pneumoniae by traditional culture techniques are low. To study the sensitivity and specificity of two different DNA extraction methods to amplify the ply gene, applied to three different types of blood culture broths, experimentally inoculated with S. pneumoniae. DNA was extracted from the cultures using an organic method or a technique that consists in dilution, washing with NaOH and concentration of the sample. This was followed by PCR amplification of a 355 pb fragment of the pneumolysin gene (ply). The organic DNA extraction method inhibited the PCR reaction at all concentrations studied (0.6 to 10(6) colony forming units/mL). Using the NaOH extraction, ply gene amplification was positive in all three blood culture broths, but only at concentrations of 10(3) colony forming units/mL, or higher. Using the same DNA extraction method, PCR was negative when the broths were inoculated with seven other related bacterial species, which results in a 100% specificity. Detection of S. pneumoniae by amplification of ply gene from blood cultures using the protocol of NaOH for DNA extraction is specific and provides results in a short lapse. However, the diagnostic sensitivity is not optimal, which limits its clinical use.

Microbial monitoring of spacecraft and associated environments

NASA Technical Reports Server (NTRS)

La Duc, M. T.; Kern, R.; Venkateswaran, K.

2004-01-01

Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore-specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained. Copyright 2004 Springer-Verlag.

Commodity-Free Calibration

NASA Technical Reports Server (NTRS)

2008-01-01

Commodity-free calibration is a reaction rate calibration technique that does not require the addition of any commodities. This technique is a specific form of the reaction rate technique, where all of the necessary reactants, other than the sample being analyzed, are either inherent in the analyzing system or specifically added or provided to the system for a reason other than calibration. After introduction, the component of interest is exposed to other reactants or flow paths already present in the system. The instrument detector records one of the following to determine the rate of reaction: the increase in the response of the reaction product, a decrease in the signal of the analyte response, or a decrease in the signal from the inherent reactant. With this data, the initial concentration of the analyte is calculated. This type of system can analyze and calibrate simultaneously, reduce the risk of false positives and exposure to toxic vapors, and improve accuracy. Moreover, having an excess of the reactant already present in the system eliminates the need to add commodities, which further reduces cost, logistic problems, and potential contamination. Also, the calculations involved can be simplified by comparison to those of the reaction rate technique. We conducted tests with hypergols as an initial investigation into the feasiblility of the technique.

Uranium Detection - Technique Validation Report

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

Colletti, Lisa Michelle; Garduno, Katherine; Lujan, Elmer J.

As a LANL activity for DOE/NNSA in support of SHINE Medical Technologies™ ‘Accelerator Technology’ we have been investigating the application of UV-vis spectroscopy for uranium analysis in solution. While the technique has been developed specifically for sulfate solutions, the proposed SHINE target solutions, it can be adapted to a range of different solution matrixes. The FY15 work scope incorporated technical development that would improve accuracy, specificity, linearity & range, precision & ruggedness, and comparative analysis. Significant progress was achieved throughout FY 15 addressing these technical challenges, as is summarized in this report. In addition, comparative analysis of unknown samples usingmore » the Davies-Gray titration technique highlighted the importance of controlling temperature during analysis (impacting both technique accuracy and linearity/range). To fully understand the impact of temperature, additional experimentation and data analyses were performed during FY16. The results from this FY15/FY16 work were presented in a detailed presentation, LA-UR-16-21310, and an update of this presentation is included with this short report summarizing the key findings. The technique is based on analysis of the most intense U(VI) absorbance band in the visible region of the uranium spectra in 1 M H 2SO 4, at λ max = 419.5 nm.« less

A novel task specific magnetic polymeric ionic liquid for selective preconcentration of potassium in oil samples using centrifuge-less dispersive liquid-liquid microextraction technique and its determination by flame atomic emission spectroscopy.

PubMed

Beiraghi, Asadollah; Shokri, Masood

2018-02-01

In the present study a new centrifuge-less dispersive liquid-liquid microextraction technique based on application of a new task specific magnetic polymeric ionic liquid (TSMPIL) as a chelating and extraction solvent for selective preconcentration of trace amounts of potassium from oil samples is developed, for the first time. After extraction, the fine droplets of TSMPIL were transferred into an eppendorf tube and diluted to 500µL using distilled water. Then, the enriched analyte was determined by flame atomic emission spectroscopy (FAES). Several important factors affecting both the complexation and extraction efficiency including extraction time, rate of vortex agitator, amount of carbonyl iron powder, pH of sample solution, volume of ionic liquid as well as effects of interfering species were investigated and optimized. Under the optimal conditions, the limits of detection (LOD) and quantification (LOQ) were 0.5 and 1.6µgL -1 respectively with the preconcentration factor of 128. The precision (RSD %) for seven replicate determinations at 10µgL -1 of potassium was better than 3.9%. The relative recoveries for the spiked samples were in the acceptable range of 95-104%. The results demonstrated that no remarkable interferences are created by other various ions in the determination of potassium, so that the tolerance limits (W Ion /W K ) of major cations and anions were in the range of 2500-10,000. The purposed method was successfully applied for the analysis of potassium in some oil samples. Copyright © 2017 Elsevier B.V. All rights reserved.

From tails to toes: developing nonlethal tissue indicators of mercury exposure in five amphibian species

USGS Publications Warehouse

Pfleeger, Adam Z.; Eagles-Smith, Collin A.; Kowalski, Brandon M.; Herring, Garth; Willacker, James J.; Jackson, Allyson K.; Pierce, John

2016-01-01

Exposure to environmental contaminants has been implicated as a factor in global amphibian decline. Mercury (Hg) is a particularly widespread contaminant that biomagnifies in amphibians and can cause a suite of deleterious effects. However, monitoring contaminant exposure in amphibian tissues may conflict with conservation goals if lethal take is required. Thus, there is a need to develop non-lethal tissue sampling techniques to quantify contaminant exposure in amphibians. Some minimally invasive sampling techniques, such as toe-clipping, are common in population-genetic research, but it is unclear if these methods can adequately characterize contaminant exposure. We examined the relationships between mercury (Hg) concentrations in non-lethally sampled tissues and paired whole-bodies in five amphibian species. Specifically, we examined the utility of three different tail-clip sections from four salamander species and toe-clips from one anuran species. Both tail and toe-clips accurately predicted whole-body THg concentrations, but the relationships differed among species and the specific tail-clip section or toe that was used. Tail-clips comprised of the distal 0–2 cm segment performed the best across all salamander species, explaining between 82 and 92 % of the variation in paired whole-body THg concentrations. Toe-clips were less effective predictors of frog THg concentrations, but THg concentrations in outer rear toes accounted for up to 79 % of the variability in frog whole-body THg concentrations. These findings suggest non-lethal sampling of tails and toes has potential applications for monitoring contaminant exposure and risk in amphibians, but care must be taken to ensure consistent collection and interpretation of samples.

Exhaled breath condensate methods adapted from human studies using longitudinal metabolomics for predicting early health alterations in dolphins.

PubMed

Borras, Eva; Aksenov, Alexander A; Baird, Mark; Novick, Brittany; Schivo, Michael; Zamuruyev, Konstantin O; Pasamontes, Alberto; Parry, Celeste; Foutouhi, Soraya; Venn-Watson, Stephanie; Weimer, Bart C; Davis, Cristina E

2017-11-01

Monitoring health conditions is essential to detect early asymptomatic stages of a disease. To achieve this, blood, urine and breath samples are commonly used as a routine clinical diagnostic. These samples offer the opportunity to detect specific metabolites related to diseases and provide a better understanding of their development. Although blood samples are commonly used routinely to monitor health, the implementation of a relatively noninvasive technique, such as exhaled breath condensate (EBC) analysis, may further benefit the well-being of both humans and other animals. EBC analysis can be used to track possible physical or biochemical alterations caused by common diseases of the bottlenose dolphin (Tursiops truncatus), such as infections or inflammatory-mediated processes. We have used an untargeted metabolomic method with liquid chromatography-mass spectrometry analysis of EBC samples to determine biomarkers related to disease development. In this study, five dolphins under human care were followed up for 1 year. We collected paired blood, physical examination information, and EBC samples. We then statistically correlated this information to predict specific health alterations. Three dolphins provided promising case study information about biomarkers related to cutaneous infections, respiratory infections, dental disease, or hormonal changes (pregnancy). The use of complementary liquid chromatography platforms, with hydrophilic interaction chromatography and reverse-phased columns, allowed us to detect a wide spectrum of EBC biomarker compounds that could be related to these health alterations. Moreover, these two analytical techniques not only provided complementary metabolite information but in both cases they also provided promising diagnostic information for these health conditions. Graphical abstract Collection of the exhaled condensed breath from a bottlenose dolphin from U.S. Navy Marine Mammal Program (MMP).

The determination of ethanol in blood and urine by mass fragmentography

NASA Technical Reports Server (NTRS)

Pereira, W. E.; Summons, R. E.; Rindfleisch, T. C.; Duffield, A. M.

1974-01-01

A mass fragmentographic technique for a rapid, specific and sensitive determination of ethanol in blood and urine is described. A Varian gas chromatograph coupled through an all-glass membrane separator to a Finnigan quadripole mass spectrometer and interfaced to a computer system is used for ethanol determination in blood and urine samples. A procedure for plotting calibration curves for ethanol quantitation is also described. Quantitation is achieved by plotting the peak area ratios of undeuterated-to-deuterated ethanol fragment ions against the amount of ethanol added. Representative results obtained by this technique are included.

Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.

PubMed

Burgess, B A; Noyes, N R; Bolte, D S; Hyatt, D R; van Metre, D C; Morley, P S

2015-01-01

Salmonella enterica is the most commonly reported cause of outbreaks of nosocomial infections in large animal veterinary teaching hospitals and the closure of equine hospitals. Rapid detection may facilitate effective control practices in equine populations. Shipping and laboratory testing typically require ≥48 h to obtain results. Lateral flow immunoassays developed for use in food-safety microbiology provide an alternative that has not been evaluated for use with faeces or environmental samples. We aimed to identify enrichment methods that would allow commercially available rapid Salmonella detection systems (lateral flow immunoassays) to be used in clinical practice with equine faecal and environmental samples, providing test results in 18-24 h. In vitro experiment. Equine faecal and environmental samples were inoculated with known quantities of S. enterica serotype Typhimurium and cultured using 2 different enrichment techniques for faeces and 4 enrichment techniques for environmental samples. Samples were tested blindly using 2 different lateral flow immunoassays and plated on agar media for confirmatory testing. In general, commercial lateral flow immunoassays resulted in fewer false-negative test results with enrichment of 1 g faecal samples in tetrathionate for 18 h, while all environmental sample enrichment techniques resulted in similar detection rates. The limit of detection from spiked samples, ∼4 colony-forming units/g, was similar for all methods evaluated. The lateral flow immunoassays evaluated could reliably detect S. enterica within 18 h, indicating that they may be useful for rapid point-of-care testing in equine practice applications. Additional evaluation is needed using samples from naturally infected cases and the environment to gain an accurate estimate of test sensitivity and specificity and to substantiate further the true value of these tests in clinical practice. © 2014 EVJ Ltd.

454 next generation-sequencing outperforms allele-specific PCR, Sanger sequencing, and pyrosequencing for routine KRAS mutation analysis of formalin-fixed, paraffin-embedded samples

PubMed Central

Altimari, Annalisa; de Biase, Dario; De Maglio, Giovanna; Gruppioni, Elisa; Capizzi, Elisa; Degiovanni, Alessio; D’Errico, Antonia; Pession, Annalisa; Pizzolitto, Stefano; Fiorentino, Michelangelo; Tallini, Giovanni

2013-01-01

Detection of KRAS mutations in archival pathology samples is critical for therapeutic appropriateness of anti-EGFR monoclonal antibodies in colorectal cancer. We compared the sensitivity, specificity, and accuracy of Sanger sequencing, ARMS-Scorpion (TheraScreen®) real-time polymerase chain reaction (PCR), pyrosequencing, chip array hybridization, and 454 next-generation sequencing to assess KRAS codon 12 and 13 mutations in 60 nonconsecutive selected cases of colorectal cancer. Twenty of the 60 cases were detected as wild-type KRAS by all methods with 100% specificity. Among the 40 mutated cases, 13 were discrepant with at least one method. The sensitivity was 85%, 90%, 93%, and 92%, and the accuracy was 90%, 93%, 95%, and 95% for Sanger sequencing, TheraScreen real-time PCR, pyrosequencing, and chip array hybridization, respectively. The main limitation of Sanger sequencing was its low analytical sensitivity, whereas TheraScreen real-time PCR, pyrosequencing, and chip array hybridization showed higher sensitivity but suffered from the limitations of predesigned assays. Concordance between the methods was k = 0.79 for Sanger sequencing and k > 0.85 for the other techniques. Tumor cell enrichment correlated significantly with the abundance of KRAS-mutated deoxyribonucleic acid (DNA), evaluated as ΔCt for TheraScreen real-time PCR (P = 0.03), percentage of mutation for pyrosequencing (P = 0.001), ratio for chip array hybridization (P = 0.003), and percentage of mutation for 454 next-generation sequencing (P = 0.004). Also, 454 next-generation sequencing showed the best cross correlation for quantification of mutation abundance compared with all the other methods (P < 0.001). Our comparison showed the superiority of next-generation sequencing over the other techniques in terms of sensitivity and specificity. Next-generation sequencing will replace Sanger sequencing as the reference technique for diagnostic detection of KRAS mutation in archival tumor tissues. PMID:23950653

Update to core reporting practices in structural equation modeling.

PubMed

Schreiber, James B

This paper is a technical update to "Core Reporting Practices in Structural Equation Modeling." 1 As such, the content covered in this paper includes, sample size, missing data, specification and identification of models, estimation method choices, fit and residual concerns, nested, alternative, and equivalent models, and unique issues within the SEM family of techniques. Copyright © 2016 Elsevier Inc. All rights reserved.

Empirical Validation of Indices for Consideration in the Revision of Recommended Senior School Financial Accounting Textbooks in Southwestern Nigeria

ERIC Educational Resources Information Center

Oyetoro, Oyebode Stephen; Ojo, Oloyede Ezekiel

2017-01-01

The study determined a significant difference in teachers' overall evaluations of six recommended Financial Accounting Textbooks in Southwestern Nigeria. It also assessed the specific evaluation parameters that account for the difference. It adopted the survey research design. The multistage sampling technique was used to select a total of 80…

Logical Framework of Forensic Identification: Ability to Resist Fabricated DNA.

PubMed

Wang, Zheng; Zhou, Di; Zhang, Suhua; Bian, Yingnan; Hu, Zhen; Zhu, Ruxin; Lu, Daru; Li, Chengtao

2015-12-01

Over the past 30 years, DNA analysis has revolutionized forensic science and has become the most useful single tool in the multifaceted fight against crime. Today, DNA profiling with sets of highly polymorphic autosomal short tandem repeat markers is widely employed and accepted in the courts due to its high discriminating power and reliability. However, an artificial bloodstain purposefully created using molecular biology techniques succeeded in tricking a leading forensic DNA laboratory. The disturbing possibility that a forensic DNA profile can be faked shocked the general public and the mass media, and generated serious discussion about the credibility of DNA evidence. Herein, we present two exemplary assays based on tissue-specific methylation patterns and cell-specific mRNA expression, respectively. These two assays can be integrated into the DNA analysis pipelines without consumption of additional samples. We show that the two assays can not only distinguish between artificial and genuine samples, but also provide information on tissue origin. The two assays were tested on natural and artificial bloodstains (generated by polymerase chain reaction and whole genome amplification technique) and the results illustrated that the logical framework of forensic identification is still useful for forensic identification with the high credibility.

Heterogeneity and frequency of BRAF mutations in primary melanoma: Comparison between molecular methods and immunohistochemistry

PubMed Central

Bruno, William; Martinuzzi, Claudia; Andreotti, Virginia; Pastorino, Lorenza; Spagnolo, Francesco; Dalmasso, Bruna; Cabiddu, Francesco; Gualco, Marina; Ballestrero, Alberto; Bianchi-Scarrà, Giovanna; Queirolo, Paola

2017-01-01

Finding the best technique to identify BRAF mutations with a high sensitivity and specificity is mandatory for accurate patient selection for target therapy. BRAF mutation frequency ranges from 40 to 60% depending on melanoma clinical characteristics and detection technique used. Intertumoral heterogeneity could lead to misinterpretation of BRAF mutational status; this is especially important if testing is performed on primary specimens, when metastatic lesions are unavailable. Aim of this study was to identify the best combination of methods for detecting BRAF mutations (among peptide nucleic acid – PNA-clamping real-time PCR, immunohistochemistry and capillary sequencing) and investigate BRAF mutation heterogeneity in a series of 100 primary melanomas and a subset of 25 matched metastatic samples. Overall, we obtained a BRAF mutation frequency of 62%, based on the combination of at least two techniques. Concordance between mutation status in primary and metastatic tumor was good but not complete (67%), when agreement of at least two techniques were considered. Next generation sequencing was used to quantify the threshold of detected mutant alleles in discordant samples. Combining different methods excludes that the observed heterogeneity is technique-based. We propose an algorithm for BRAF mutation testing based on agreement between immunohistochemistry and PNA; a third molecular method could be added in case of discordance of the results. Testing the primary tumor when the metastatic sample is unavailable is a good option if at least two methods of detection are used, however the presence of intertumoral heterogeneity or the occurrence of additional primaries should be carefully considered. PMID:28039443

Discussing State-of-the-Art Spatial Visualization Techniques Applicable for the Epidemiological Surveillance Data on the Example of Campylobacter spp. in Raw Chicken Meat.

PubMed

Plaza-Rodríguez, C; Appel, B; Kaesbohrer, A; Filter, M

2016-08-01

Within the European activities for the 'Monitoring and Collection of Information on Zoonoses', annually EFSA publishes a European report, including information related to the prevalence of Campylobacter spp. in Germany. Spatial epidemiology becomes here a fundamental tool for the generation of these reports, including the representation of prevalence as an essential element. Until now, choropleth maps are the default visualization technique applied in epidemiological monitoring and surveillance reports made by EFSA and German authorities. However, due to its limitations, it seems to be reasonable to explore alternative chart type. Four maps including choropleth, cartogram, graduated symbols and dot-density maps were created to visualize real-world sample data on the prevalence of Campylobacter spp. in raw chicken meat samples in Germany in 2011. In addition, adjacent and coincident maps were created to visualize also the associated uncertainty. As an outcome, we found that there is not a single data visualization technique that encompasses all the necessary features to visualize prevalence data alone or prevalence data together with their associated uncertainty. All the visualization techniques contemplated in this study demonstrated to have both advantages and disadvantages. To determine which visualization technique should be used for future reports, we recommend to create a dialogue between end-users and epidemiologists on the basis of sample data and charts. The final decision should also consider the knowledge and experience of end-users as well as the specific objective to be achieved with the charts. © 2015 The Authors. Zoonoses and Public Health Published by Blackwell Verlag GmbH.

Label-free imaging of trabecular meshwork cells using Coherent Anti-Stokes Raman Scattering (CARS) microscopy

PubMed Central

Lei, Tim C.; Ammar, David A.; Masihzadeh, Omid; Gibson, Emily A.

2011-01-01

Purpose To image the human trabecular meshwork (TM) using a non-invasive, non-destructive technique without the application of exogenous label. Methods Flat-mounted TM samples from a human cadaver eye were imaged using two nonlinear optical techniques: coherent anti-Stokes Raman scattering (CARS) and two-photon autofluorescence (TPAF). In TPAF, two optical photons are simultaneously absorbed and excite molecules in the sample that then emit a higher energy photon. The signal is predominately from collagen and elastin. The CARS technique uses two laser frequencies to specifically excite carbon-hydrogen bonds, allowing the visualization of lipid-rich cell membranes. Multiple images were taken along an axis perpendicular to the surface of the TM for subsequent analysis. Results Analysis of multiple TPAF images of the TM reveals the characteristic overlapping bundles of collagen of various sizes. Simultaneous CARS imaging revealed elliptical structures of ~7×10 µm in diameter populating the meshwork which were consistent with TM cells. Irregularly shaped objects of ~4 µm diameter appeared in both the TPAF and CARS channels, and are consistent with melanin granules. Conclusions CARS techniques were successful in imaging live TM cells in freshly isolated human TM samples. Similar images have been obtained with standard histological techniques, however the method described here has the advantage of being performed on unprocessed, unfixed tissue free from the potential distortions of the fine tissue morphology that can occur due to infusion of fixatives and treatment with alcohols. CARS imaging of the TM represents a new avenue for exploring details of aqueous outflow and TM cell physiology. PMID:22025898

Improved removal of blood contamination from ThinPrep cervical cytology samples for Raman spectroscopic analysis.

PubMed

Traynor, Damien; Duraipandian, Shiyamala; Martin, Cara M; O'Leary, John J; Lyng, Fiona M

2018-05-01

There is an unmet need for methods to help in the early detection of cervical precancer. Optical spectroscopy-based techniques, such as Raman spectroscopy, have shown great potential for diagnosis of different cancers, including cervical cancer. However, relatively few studies have been carried out on liquid-based cytology (LBC) pap test specimens and confounding factors, such as blood contamination, have been identified. Previous work reported a method to remove blood contamination before Raman spectroscopy by pretreatment of the slides with hydrogen peroxide. The aim of the present study was to extend this work to excessively bloody samples to see if these could be rendered suitable for Raman spectroscopy. LBC ThinPrep specimens were treated by adding hydrogen peroxide directly to the vial before slide preparation. Good quality Raman spectra were recorded from negative and high grade (HG) cytology samples with no blood contamination and with heavy blood contamination. Good classification between negative and HG cytology could be achieved for samples with no blood contamination (sensitivity 92%, specificity 93%) and heavy blood contamination (sensitivity 89%, specificity 88%) with poorer classification when samples were combined (sensitivity 82%, specificity 87%). This study demonstrates for the first time the improved potential of Raman spectroscopy for analysis of ThinPrep specimens regardless of blood contamination. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Quantum dots for a high-throughput Pfu polymerase based multi-round polymerase chain reaction (PCR).

PubMed

Sang, Fuming; Zhang, Zhizhou; Yuan, Lin; Liu, Deli

2018-02-26

Multi-round PCR is an important technique for obtaining enough target DNA from rare DNA resources, and is commonly used in many fields including forensic science, ancient DNA analysis and cancer research. However, multi-round PCR is often aborted, largely due to the accumulation of non-specific amplification during repeated amplifications. Here, we developed a Pfu polymerase based multi-round PCR technique assisted by quantum dots (QDs). Different PCR assays, DNA polymerases (Pfu and Taq), DNA sizes and GC amounts were compared in this study. In the presence of QDs, PCR specificity could be retained even in the ninth-round amplification. Moreover, the longer and more complex the targets were, the earlier the abortion happened in multi-round PCR. However, no obvious enhancement of specificity was found in multi-round PCR using Taq DNA polymerase. Significantly, the fidelity of Pfu polymerase based multi-round PCR was not sacrificed in the presence of QDs. Besides, pre-incubation at 50 °C for an hour had no impact on multi-round PCR performance, which further authenticated the hot start effect of QDs modulated in multi-round PCR. The findings of this study demonstrated that a cost-effective and promising multi-round PCR technique for large-scale and high-throughput sample analysis could be established with high specificity, sensibility and accuracy.

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

Timmermans, F. J.; Otto, C.

New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemicallymore » or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy.« less

Contributed review: Review of integrated correlative light and electron microscopy.

PubMed

Timmermans, F J; Otto, C

2015-01-01

New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemically or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy.

Nitrogen doped activated carbon from pea skin for high performance supercapacitor

NASA Astrophysics Data System (ADS)

Ahmed, Sultan; Ahmed, Ahsan; Rafat, M.

2018-04-01

In this work, nitrogen doped porous carbon (NDC) has been synthesized employing a facile two-step process. Firstly, carbon precursor (pea skin) was heated with melamine (acting as nitrogen source) followed by activation with KOH in different ratios. The dependence of porosity and nitrogen content on impregnation ratio was extensively studied. Other textural properties of prepared NDC sample were studied using standard techniques of material characterization. The electrochemical performance of NDC sample as an electrode was studied in two-electrode symmetric supercapacitor system. 1 M LiTFSI (lithium bis-trifluoromethanesulfonimide) solution in IL EMITFSI (1-Ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide), was used as electrolyte. It was found that the fabricated supercapacitor cell offers high values of specific capacitance (141.1 F g‑1), specific energy (19.6 Wh kg‑1) and specific power (25.4 kW kg‑1) at current density of 1.3 A g‑1. More importantly, the fabricated supercapacitor cell shows capacitance retention of ∼75%, for more than 5000 cycles. The enhanced performance of NDC sample is primarily due to large surface area with favorable surface structure (contributing to double layer capacitance) and presence of nitrogen functionalities (contributing to pseudo-capacitance). Such important features make the synthesized NDC sample, an attractive choice for electrode material in high performance supercapacitor.

Sensitivity and specificity of the circulating cathodic antigen rapid urine test in the diagnosis of Schistosomiasis mansoni infection and evaluation of morbidity in a low- endemic area in Brazil.

PubMed

Ferreira, Fernanda Teixeira; Fidelis, Thiago André; Pereira, Thiago Almeida; Otoni, Alba; Queiroz, Leonardo Campos; Amâncio, Frederico Figueiredo; Antunes, Carlos Maurício; Lambertucci, José Roberto

2017-01-01

The Kato-Katz technique is the standard diagnostic test for Schistosoma mansoni infection in rural areas. However, the utility of this method is severely limited by the day-to-day variability in host egg excretion in the stool. In high-transmission areas, the point-of-care circulating cathodic antigen (POC-CCA) urine assay has proven to be a reliable test. However, investigations of the reliability of the POC-CCA assay in low-transmission regions are under way. This study aimed to evaluate the sensitivity and specificity of the POC-CCA assay and the morbidity of schistosomiasis in a low-endemic area in Brazil. Pains City is a low-transmission zone for schistosomiasis. A total of 300 subjects aged 7-76 years were randomly selected for the POC-CCA cassette test. For S. mansoni diagnosis, three stool samples on six slides were compared with one urine sample for each subject. The sensitivity and specificity in the absence of a gold standard were calculated using latent class analysis. Clinical examinations and abdominal ultrasounds were performed in 181 volunteers to evaluate morbidity associated with schistosomiasis. The sensitivity and specificity of the Kato-Katz technique were 25.6% and 94.6%, respectively. By contrast, the sensitivity and specificity of the POC-CCA assay were 68.1% and 72.8%, respectively. Hepatosplenic schistosomiasis was diagnosed in two patients (1.1%). Overall, the POC-CCA urine assay proved to be a useful test for diagnosing S. mansoni in a low-endemic area in Brazil. Severe clinical forms of schistosomiasis can be present even in such low-endemic areas.

Optical biopsy of head and neck cancer using hyperspectral imaging and convolutional neural networks

NASA Astrophysics Data System (ADS)

Halicek, Martin; Little, James V.; Wang, Xu; Patel, Mihir; Griffith, Christopher C.; El-Deiry, Mark W.; Chen, Amy Y.; Fei, Baowei

2018-02-01

Successful outcomes of surgical cancer resection necessitate negative, cancer-free surgical margins. Currently, tissue samples are sent to pathology for diagnostic confirmation. Hyperspectral imaging (HSI) is an emerging, non-contact optical imaging technique. A reliable optical method could serve to diagnose and biopsy specimens in real-time. Using convolutional neural networks (CNNs) as a tissue classifier, we developed a method to use HSI to perform an optical biopsy of ex-vivo surgical specimens, collected from 21 patients undergoing surgical cancer resection. Training and testing on samples from different patients, the CNN can distinguish squamous cell carcinoma (SCCa) from normal aerodigestive tract tissues with an area under the curve (AUC) of 0.82, 81% accuracy, 81% sensitivity, and 80% specificity. Additionally, normal oral tissues can be sub-classified into epithelium, muscle, and glandular mucosa using a decision tree method, with an average AUC of 0.94, 90% accuracy, 93% sensitivity, and 89% specificity. After separately training on thyroid tissue, the CNN differentiates between thyroid carcinoma and normal thyroid with an AUC of 0.95, 92% accuracy, 92% sensitivity, and 92% specificity. Moreover, the CNN can discriminate medullary thyroid carcinoma from benign multi-nodular goiter (MNG) with an AUC of 0.93, 87% accuracy, 88% sensitivity, and 85% specificity. Classical-type papillary thyroid carcinoma is differentiated from benign MNG with an AUC of 0.91, 86% accuracy, 86% sensitivity, and 86% specificity. Our preliminary results demonstrate that an HSI-based optical biopsy method using CNNs can provide multi-category diagnostic information for normal head-and-neck tissue, SCCa, and thyroid carcinomas. More patient data are needed in order to fully investigate the proposed technique to establish reliability and generalizability of the work.

Rapid analysis method for the determination of 14C specific activity in irradiated graphite

PubMed Central

Remeikis, Vidmantas; Lagzdina, Elena; Garbaras, Andrius; Gudelis, Arūnas; Garankin, Jevgenij; Juodis, Laurynas; Duškesas, Grigorijus; Lingis, Danielius; Abdulajev, Vladimir; Plukis, Artūras

2018-01-01

14C is one of the limiting radionuclides used in the categorization of radioactive graphite waste; this categorization is crucial in selecting the appropriate graphite treatment/disposal method. We propose a rapid analysis method for 14C specific activity determination in small graphite samples in the 1–100 μg range. The method applies an oxidation procedure to the sample, which extracts 14C from the different carbonaceous matrices in a controlled manner. Because this method enables fast online measurement and 14C specific activity evaluation, it can be especially useful for characterizing 14C in irradiated graphite when dismantling graphite moderator and reflector parts, or when sorting radioactive graphite waste from decommissioned nuclear power plants. The proposed rapid method is based on graphite combustion and the subsequent measurement of both CO2 and 14C, using a commercial elemental analyser and the semiconductor detector, respectively. The method was verified using the liquid scintillation counting (LSC) technique. The uncertainty of this rapid method is within the acceptable range for radioactive waste characterization purposes. The 14C specific activity determination procedure proposed in this study takes approximately ten minutes, comparing favorably to the more complicated and time consuming LSC method. This method can be potentially used to radiologically characterize radioactive waste or used in biomedical applications when dealing with the specific activity determination of 14C in the sample. PMID:29370233

Rapid analysis method for the determination of 14C specific activity in irradiated graphite.

PubMed

Remeikis, Vidmantas; Lagzdina, Elena; Garbaras, Andrius; Gudelis, Arūnas; Garankin, Jevgenij; Plukienė, Rita; Juodis, Laurynas; Duškesas, Grigorijus; Lingis, Danielius; Abdulajev, Vladimir; Plukis, Artūras

2018-01-01

14C is one of the limiting radionuclides used in the categorization of radioactive graphite waste; this categorization is crucial in selecting the appropriate graphite treatment/disposal method. We propose a rapid analysis method for 14C specific activity determination in small graphite samples in the 1-100 μg range. The method applies an oxidation procedure to the sample, which extracts 14C from the different carbonaceous matrices in a controlled manner. Because this method enables fast online measurement and 14C specific activity evaluation, it can be especially useful for characterizing 14C in irradiated graphite when dismantling graphite moderator and reflector parts, or when sorting radioactive graphite waste from decommissioned nuclear power plants. The proposed rapid method is based on graphite combustion and the subsequent measurement of both CO2 and 14C, using a commercial elemental analyser and the semiconductor detector, respectively. The method was verified using the liquid scintillation counting (LSC) technique. The uncertainty of this rapid method is within the acceptable range for radioactive waste characterization purposes. The 14C specific activity determination procedure proposed in this study takes approximately ten minutes, comparing favorably to the more complicated and time consuming LSC method. This method can be potentially used to radiologically characterize radioactive waste or used in biomedical applications when dealing with the specific activity determination of 14C in the sample.

Detection of distribution of avian influenza H5N1 virus by immunohistochemistry, chromogenic in situ hybridization and real-time PCR techniques in experimentally infected chickens.

PubMed

Chamnanpood, Chanpen; Sanguansermsri, Donruedee; Pongcharoen, Sutatip; Sanguansermsri, Phanchana

2011-03-01

Ten specific pathogen free (SPF) chickens were inoculated intranasally with avian influenza virus subtype H5N1. Evaluation revealed distribution of the virus in twelve organs: liver, intestine, bursa, lung, trachea, thymus, heart, pancreas, brain, spleen, kidney, and esophagus. Immunohistochemistry (IHC), chromogenic in situ hybridization (CISH), and real-time polymerase chain reaction (PCR) were developed and compared for detection of the virus from the organs. The distribution of avian influenza H5N1 in chickens varied by animal and detecting technique. The heart, kidneys, intestines, lungs, and pancreas were positive with all three techniques, while the others varied by techique. The three techniques can be used to detect avian influenza effectively, but the pros and cons of each technique need to be determined. The decision of which technique to use depends on the objective of the examination, budget, type and quality of samples, laboratory facilities and technician skills.

Applications of Flow Cytometry to Clinical Microbiology†

PubMed Central

Álvarez-Barrientos, Alberto; Arroyo, Javier; Cantón, Rafael; Nombela, César; Sánchez-Pérez, Miguel

2000-01-01

Classical microbiology techniques are relatively slow in comparison to other analytical techniques, in many cases due to the need to culture the microorganisms. Furthermore, classical approaches are difficult with unculturable microorganisms. More recently, the emergence of molecular biology techniques, particularly those on antibodies and nucleic acid probes combined with amplification techniques, has provided speediness and specificity to microbiological diagnosis. Flow cytometry (FCM) allows single- or multiple-microbe detection in clinical samples in an easy, reliable, and fast way. Microbes can be identified on the basis of their peculiar cytometric parameters or by means of certain fluorochromes that can be used either independently or bound to specific antibodies or oligonucleotides. FCM has permitted the development of quantitative procedures to assess antimicrobial susceptibility and drug cytotoxicity in a rapid, accurate, and highly reproducible way. Furthermore, this technique allows the monitoring of in vitro antimicrobial activity and of antimicrobial treatments ex vivo. The most outstanding contribution of FCM is the possibility of detecting the presence of heterogeneous populations with different responses to antimicrobial treatments. Despite these advantages, the application of FCM in clinical microbiology is not yet widespread, probably due to the lack of access to flow cytometers or the lack of knowledge about the potential of this technique. One of the goals of this review is to attempt to mitigate this latter circumstance. We are convinced that in the near future, the availability of commercial kits should increase the use of this technique in the clinical microbiology laboratory. PMID:10755996

Timothy-specific IgG antibody levels vary with the pollen seasons.

PubMed

Nordvall, S L; Larsson, P H; Johansson, S G

1986-11-01

Serum samples were collected from eight grass pollen hypersensitive children during a 4-year period. The sera were assayed for contents of timothy-specific IgE antibodies by RAST. Timothy-specific IgG and IgA antibodies were quantified by a refined ELISA in which covalent binding of the antigen to the polystyrene solid phase had been performed. IgG antibodies were also assayed by a Sepharose-protein-A technique with radiolabelled timothy allergens as the antigen. It was possible to register clearcut seasonal variations with postseasonally boosted antibody levels not only of timothy-specific IgE but also of IgG antibody. Both IgG1 and IgG4 antibodies specific for timothy showed seasonal variations of a similar degree. It was not possible to register seasonal variations of the same magnitude of timothy-specific IgA antibodies.

Direct detection of fibrinogen in human plasma using electric-double-layer gated AlGaN/GaN high electron mobility transistors

NASA Astrophysics Data System (ADS)

Regmi, Abiral; Sarangadharan, Indu; Chen, Yen-Wen; Hsu, Chen-Pin; Lee, Geng-Yen; Chyi, Jen-Inn; Shiesh, Shu-Chu; Lee, Gwo-Bin; Wang, Yu-Lin

2017-08-01

Fibrinogen found in blood plasma is an important protein biomarker for potentially fatal diseases such as cardiovascular diseases. This study focuses on the development of an assay to detect plasmatic fibrinogen using electrical double layer gated AlGaN/GaN high electron mobility transistor biosensors without complex sample pre-treatment methods used in the traditional assays. The test results in buffer solution and clinical plasma samples show high sensitivity, specificity, and dynamic range. The sensor exhibits an ultra-low detection limit of 0.5 g/l and a detection range of 0.5-4.5 g/l in 1× PBS with 1% BSA. The concentration dependent sensor signal in human serum samples demonstrates the specificity to fibrinogen in a highly dense matrix of background proteins. The sensor does not require complicated automation, and quantitative results are obtained in 5 min with <5 μl sample volume. This sensing technique is ideal for speedy blood based diagnostics such as POC (point of care) tests, homecare tests, or personalized healthcare.

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

Amblard, A.; Riguccini, L.; Temi, P.

We compute the properties of a sample of 221 local, early-type galaxies with a spectral energy distribution (SED) modeling software, CIGALEMC. Concentrating on the star-forming (SF) activity and dust contents, we derive parameters such as the specific star formation rate (sSFR), the dust luminosity, dust mass, and temperature. In our sample, 52% is composed of elliptical (E) galaxies and 48% of lenticular (S0) galaxies. We find a larger proportion of S0 galaxies among galaxies with a large sSFR and large specific dust emission. The stronger activity of S0 galaxies is confirmed by larger dust masses. We investigate the relative proportionmore » of active galactic nuclei (AGNs) and SF galaxies in our sample using spectroscopic Sloan Digital Sky Survey data and near-infrared selection techniques, and find a larger proportion of AGN-dominated galaxies in the S0 sample than the E one. This could corroborate a scenario where blue galaxies evolve into red ellipticals by passing through an S0 AGN active period while quenching its star formation. Finally, we find a good agreement comparing our estimates with color indicators.« less

Sampling Approaches for Multi-Domain Internet Performance Measurement Infrastructures

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

Calyam, Prasad

2014-09-15

The next-generation of high-performance networks being developed in DOE communities are critical for supporting current and emerging data-intensive science applications. The goal of this project is to investigate multi-domain network status sampling techniques and tools to measure/analyze performance, and thereby provide “network awareness” to end-users and network operators in DOE communities. We leverage the infrastructure and datasets available through perfSONAR, which is a multi-domain measurement framework that has been widely deployed in high-performance computing and networking communities; the DOE community is a core developer and the largest adopter of perfSONAR. Our investigations include development of semantic scheduling algorithms, measurement federationmore » policies, and tools to sample multi-domain and multi-layer network status within perfSONAR deployments. We validate our algorithms and policies with end-to-end measurement analysis tools for various monitoring objectives such as network weather forecasting, anomaly detection, and fault-diagnosis. In addition, we develop a multi-domain architecture for an enterprise-specific perfSONAR deployment that can implement monitoring-objective based sampling and that adheres to any domain-specific measurement policies.« less

Heterodyne method for high specificity gas detection.

NASA Technical Reports Server (NTRS)

Dimeff, J.; Donaldson, R. W.; Gunter, W. D., Jr.; Jaynes, D. N.; Margozzi, A. P.; Deboo, G. J.; Mcclatchie, E. A.; Williams, K. G.

1971-01-01

This paper describes a new technique for measuring trace quantities of gases. The technique involves the use of a reference cell (containing a known amount of the gas being sought) and a sample cell (containing an unknown amount of the same gas) wherein the gas densities are modulated. Light passing through the two cells in sequence is modulated in intensity at the vibrational-rotational lines characteristic of the absorption spectrum for the gas of interest. Since the absorption process is nonlinear, modulating the two absorption cells at two different frequencies gives rise to a heterodyning effect, which in turn introduces sum and difference frequencies in the detected signal. Measuring the ratio of the difference frequency signal for example, to the signal introduced by the reference cell provides a normalized measure of the amount of the gas in the sample cell. The readings produced are thereby independent of source intensity, window transparency, and detector sensitivity. Experimental evaluation of the technique suggests that it should be applicable to a wide range of gases, that it should be able to reject spurious signals due to unwanted gases, and that it should be sensitive to concentrations of the order of 10 to the minus 8th power when used with a sample cell of only 20 cm length.

Assessment of the Authenticity of Herbal Dietary Supplements: Comparison of Chemical and DNA Barcoding Methods.

PubMed

Pawar, Rahul S; Handy, Sara M; Cheng, Raymond; Shyong, Nicole; Grundel, Erich

2017-07-01

About 7 % of the U. S. population reports using botanical dietary supplements. Increased use of such supplements has led to discussions related to their authenticity and quality. Reports of adulteration with substandard materials or pharmaceuticals are of concern because such substitutions, whether inadvertent or deliberate, may reduce the efficacy of specific botanicals or lead to adverse events. Methods for verifying the identity of botanicals include macroscopic and microscopic examinations, chemical analysis, and DNA-based methods including DNA barcoding. Macroscopic and microscopic examinations may fail when a supplement consists of botanicals that have been processed beyond the ability to provide morphological characterizations. Chemical analysis of specific marker compounds encounters problems when these compounds are not distinct to a given species or when purified reference standards are not available. Recent investigations describing DNA barcoding analysis of botanical dietary supplements have raised concerns about the authenticity of the supplements themselves as well as the appropriateness of using DNA barcoding techniques with finished botanical products. We collected 112 market samples of frequently consumed botanical dietary supplements of ginkgo, soy, valerian, yohimbe, and St. John's wort and analyzed each for specific chemical markers (i.e., flavonol glycosides, total isoflavones, total valerenic acids, yohimbine, and hypericins, respectively). We used traditional DNA barcoding techniques targeting the nuclear ITS2 gene and the chloroplast gene psb A- trn H on the same samples to determine the presence of DNA of the labelled ingredient. We compared the results obtained by both methods to assess the contribution of each in determining the identity of the samples. Georg Thieme Verlag KG Stuttgart · New York.

Quantitative analysis of aberrant protein glycosylation in liver cancer plasma by AAL-enrichment and MRM mass spectrometry.

PubMed

Ahn, Yeong Hee; Shin, Park Min; Kim, Yong-Sam; Oh, Na Ree; Ji, Eun Sun; Kim, Kwang Hoe; Lee, Yeon Jung; Kim, Sung Ho; Yoo, Jong Shin

2013-11-07

A lectin-coupled mass spectrometry (MS) approach was employed to quantitatively monitor aberrant protein glycosylation in liver cancer plasma. To do this, we compared the difference in the total protein abundance of a target glycoprotein between hepatocellular carcinoma (HCC) plasmas and hepatitis B virus (HBV) plasmas, as well as the difference in lectin-specific protein glycoform abundance of the target glycoprotein. Capturing the lectin-specific protein glycoforms from a plasma sample was accomplished by using a fucose-specific aleuria aurantia lectin (AAL) immobilized onto magnetic beads via a biotin-streptavidin conjugate. Following tryptic digestion of both the total plasma and its AAL-captured fraction of each HCC and HBV sample, targeted proteomic mass spectrometry was conducted quantitatively by a multiple reaction monitoring (MRM) technique. From the MRM-based analysis of the total plasmas and AAL-captured fractions, differences between HCC and HBV plasma groups in fucosylated glycoform levels of target glycoproteins were confirmed to arise from both the change in the total protein abundance of the target proteins and the change incurred by aberrant fucosylation on target glycoproteins in HCC plasma, even when no significant change occurs in the total protein abundance level. Combining the MRM-based analysis method with the lectin-capturing technique proved to be a successful means of quantitatively investigating aberrant protein glycosylation in cancer plasma samples. Additionally, it was elucidated that the differences between HCC and control groups in fucosylated biomarker candidates A1AT and FETUA mainly originated from an increase in fucosylation levels on these target glycoproteins, rather than an increase in the total protein abundance of the target glycoproteins.

Differences in head impulse test results due to analysis techniques.

PubMed

Cleworth, Taylor W; Carpenter, Mark G; Honegger, Flurin; Allum, John H J

2017-01-01

Different analysis techniques are used to define vestibulo-ocular reflex (VOR) gain between eye and head angular velocity during the video head impulse test (vHIT). Comparisons would aid selection of gain techniques best related to head impulse characteristics and promote standardisation. Compare and contrast known methods of calculating vHIT VOR gain. We examined lateral canal vHIT responses recorded from 20 patients twice within 13 weeks of acute unilateral peripheral vestibular deficit onset. Ten patients were tested with an ICS Impulse system (GN Otometrics) and 10 with an EyeSeeCam (ESC) system (Interacoustics). Mean gain and variance were computed with area, average sample gain, and regression techniques over specific head angular velocity (HV) and acceleration (HA) intervals. Results for the same gain technique were not different between measurement systems. Area and average sample gain yielded equally lower variances than regression techniques. Gains computed over the whole impulse duration were larger than those computed for increasing HV. Gain over decreasing HV was associated with larger variances. Gains computed around peak HV were smaller than those computed around peak HA. The median gain over 50-70 ms was not different from gain around peak HV. However, depending on technique used, the gain over increasing HV was different from gain around peak HA. Conversion equations between gains obtained with standard ICS and ESC methods were computed. For low gains, the conversion was dominated by a constant that needed to be added to ESC gains to equal ICS gains. We recommend manufacturers standardize vHIT gain calculations using 2 techniques: area gain around peak HA and peak HV.

Gaussian graphical modeling reveals specific lipid correlations in glioblastoma cells

NASA Astrophysics Data System (ADS)

Mueller, Nikola S.; Krumsiek, Jan; Theis, Fabian J.; Böhm, Christian; Meyer-Bäse, Anke

2011-06-01

Advances in high-throughput measurements of biological specimens necessitate the development of biologically driven computational techniques. To understand the molecular level of many human diseases, such as cancer, lipid quantifications have been shown to offer an excellent opportunity to reveal disease-specific regulations. The data analysis of the cell lipidome, however, remains a challenging task and cannot be accomplished solely based on intuitive reasoning. We have developed a method to identify a lipid correlation network which is entirely disease-specific. A powerful method to correlate experimentally measured lipid levels across the various samples is a Gaussian Graphical Model (GGM), which is based on partial correlation coefficients. In contrast to regular Pearson correlations, partial correlations aim to identify only direct correlations while eliminating indirect associations. Conventional GGM calculations on the entire dataset can, however, not provide information on whether a correlation is truly disease-specific with respect to the disease samples and not a correlation of control samples. Thus, we implemented a novel differential GGM approach unraveling only the disease-specific correlations, and applied it to the lipidome of immortal Glioblastoma tumor cells. A large set of lipid species were measured by mass spectrometry in order to evaluate lipid remodeling as a result to a combination of perturbation of cells inducing programmed cell death, while the other perturbations served solely as biological controls. With the differential GGM, we were able to reveal Glioblastoma-specific lipid correlations to advance biomedical research on novel gene therapies.

Establishment and operation of the National Accident Sampling System (NASS) team within the cities of Ft. Lauderdale/Hollywood, Florida

NASA Astrophysics Data System (ADS)

Beddow, B.; Roberts, C.; Rankin, J.; Bloch, A.; Peizer, J.

1981-01-01

The National Accident Sampling System (NASS) is described. The study area discussed is one of the original ten sites selected for NASS implementation. In addition to collecting data from the field, the original ten sites address questions of feasibility of the plan, projected results of the data collection effort, and specific operational topics, e.g., team size, sampling requirements, training approaches, quality control procedures, and field techniques. Activities and results of the first three years of the project, for both major tasks (establishment and operation) are addressed. Topics include: study area documentation; team description, function and activities; problems and solutions; and recommendations.

Studies into the microbial spectrum of apical periodontitis.

PubMed

Brauner, A W; Conrads, G

1995-09-01

This study examined the variety of obligate and facultative anaerobic bacterial species recovered from cases of acute apical periodontitis. A total of 19 root canal samples and 24 periapical granuloma samples were taken from patients suffering pain and discomfort. Bacteria were identified by applying the following techniques: culturing on various media, Gram-staining and using commercially available biochemical test strips. In addition, Prevotella intermedia and Porphyromonas endodontalis were differentiated on a molecular genetic level using species-specific oligodeoxynucleotide probes. The most frequently identified bacteria were Prevotella intermedia, Bifidobacterium spp., Streptococcus sanguis, Streptococcus milleri-group and Bacteroides spp. Obligate anaerobes occurred at a rate of 82.3%, and the average number of isolates was 6.4 per sample.

Improvements in the gaseous hydrogen-water equilibration technique for hydrogen isotope ratio analysis

USGS Publications Warehouse

Coplen, T.B.; Wildman, J.D.; Chen, J.

1991-01-01

Improved precision in the H2-H2O equilibration method for ??D analysis has been achieved in an automated system. Reduction in 1-?? standard deviation of a single mass-spectrometer analysis to 1.3??? is achieved by (1) bonding catalyst to glass rods and assigning use to specific equilibration chambers to monitor performance of catalyst, (2) improving the apparatus design, and (3) reducing the H3+ contribution of the mass-spectrometer ion source. For replicate analysis of a water sample, the standard deviation improved to 0.8???. H2S-bearing samples and samples as small as 0.1 mL can be analyzed routinely with this method.

Investigation of Zerodur material processing

NASA Technical Reports Server (NTRS)

Johnson, R. Barry

1993-01-01

The Final Report of the Center for Applied Optics (CAO), of The University of Alabama (UAH) study entitled 'Investigation of Zerodur Material Processing' is presented. The objectives of the effort were to prepare glass samples by cutting, grinding, etching, and polishing block Zerodur to desired specifications using equipment located in the optical shop located in the Optical System Branch at NASA/MSFC; characterize samples for subsurface damage and surface roughness; utilize Zerodur samples for coating investigations; and perform investigations into enhanced optical fabrication and metrology techniques. The results of this investigation will be used to support the Advanced X Ray Astrophysics Facility (AXAF) program as well as other NASA/MSFC research programs. The results of the technical effort are presented and discussed.

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

Greenberg, Jim; Penuelas, J.; Guenther, Alex B.

To survey landscape-scale fluxes of biogenic gases, a100-meterTeflon tube was attached to a tethered balloon as a sampling inlet for a fast response Proton Transfer Reaction Mass Spectrometer (PTRMS). Along with meteorological instruments deployed on the tethered balloon and at 3-mand outputs from a regional weather model, these observations were used to estimate landscape scale biogenic volatile organic compound fluxes with two micrometeorological techniques: mixed layer variance and surface layer gradients. This highly mobile sampling system was deployed at four field sites near Barcelona to estimate landscape-scale BVOC emission factors in a relatively short period (3 weeks). The two micrometeorologicalmore » techniques agreed within the uncertainty of the flux measurements at all four sites even though the locations had considerable heterogeneity in species distribution and complex terrain. The observed fluxes were significantly different than emissions predicted with an emission model using site-specific emission factors and land-cover characteristics. Considering the wide range in reported BVOC emission factors of VOCs for individual vegetation species (more than an order of magnitude), this flux estimation technique is useful for constraining BVOC emission factors used as model inputs.« less

The growth and perfection of β-cyclotetramethylene-tetranitramine (HMX) studied by laboratory and synchrotron X-ray topography

NASA Astrophysics Data System (ADS)

Gallagher, H. G.; Sherwood, J. N.; Vrcelj, R. M.

2017-10-01

An examination has been made of the defect structure of crystals of the energetic material β-cyclotetramethylene-tetranitramine (HMX) using both Laboratory (Lang method) and Synchrotron (Bragg Reflection and Laue method) techniques. The results of the three methods are compared with particular attention to the influence of potential radiation damage caused to the samples by the latter, more energetic, technique. The comparison shows that both techniques can be confidently used to evaluate the defect structures yielding closely similar results. The results show that, even under the relatively casual preparative methods used (slow evaporation of unstirred solutions at constant temperature), HMX crystals of high perfection can be produced. The crystals show well defined bulk defect structures characteristic of organic materials in general: growth dislocations, twins, growth sector boundaries, growth banding and solvent inclusions. The distribution of the defects in specific samples is correlated with the morphological variation of the grown crystals. The results show promise for the further evaluation and characterisation of the structure and properties of dislocations and other defects and their involvement in mechanical and energetic processes in this material.

Detection of Methylated Circulating DNA as Noninvasive Biomarkers for Breast Cancer Diagnosis

PubMed Central

Cheuk, Isabella Wai Yin; Shin, Vivian Yvonne

2017-01-01

Internationally, breast cancer is the most common female cancer, and is induced by a combination of environmental, genetic, and epigenetic risk factors. Despite the advancement of imaging techniques, invasive sampling of breast epithelial cells is the only definitive diagnostic procedure for patients with breast cancer. To date, molecular biomarkers with high sensitivity and specificity for the screening and early detection of breast cancer are lacking. Recent evidence suggests that the detection of methylated circulating cell-free DNA in the peripheral blood of patients with cancer may be a promising quantitative and noninvasive method for cancer diagnosis. Methylation detection based on a multi-gene panel, rather than on the methylation status of a single gene, may be used to increase the sensitivity and specificity of breast cancer screening. In this review, the results of 14 relevant studies, investigating the efficacy of cell-free DNA methylation screening for breast cancer diagnosis, have been summarized. The genetic risk factors for breast cancer, the methods used for breast cancer detection, and the techniques and limitations related to the detection of cell-free DNA methylation status, have also been reviewed and discussed. From this review, we conclude that the analysis of peripheral blood or other samples to detect differentially methylated cell-free DNA is a promising technique for use in clinical settings, and may improve the sensitivity of screening for both, early detection and disease relapse, and thus improve the future prognosis of patients with breast cancer. PMID:28382090

Clinical diagnosis of ventilator associated pneumonia revisited: comparative validation using immediate post-mortem lung biopsies.

PubMed

Fàbregas, N; Ewig, S; Torres, A; El-Ebiary, M; Ramirez, J; de La Bellacasa, J P; Bauer, T; Cabello, H

1999-10-01

A study was undertaken to assess the diagnostic value of different clinical criteria and the impact of microbiological testing on the accuracy of clinical diagnosis of suspected ventilator associated pneumonia (VAP). Twenty five deceased mechanically ventilated patients were studied prospectively. Immediately after death, multiple bilateral lung biopsy specimens (16 specimens/patient) were obtained for histological examination and quantitative lung cultures. The presence of both histological pneumonia and positive lung cultures was used as a reference test. The presence of infiltrates on the chest radiograph and two of three clinical criteria (leucocytosis, purulent secretions, fever) had a sensitivity of 69% and a specificity of 75%; the corresponding numbers for the clinical pulmonary infection score (CPIS) were 77% and 42%. Non-invasive as well as invasive sampling techniques had comparable values. The combination of all techniques achieved a sensitivity of 85% and a specificity of 50%, and these values remained virtually unchanged despite the presence of previous treatment with antibiotics. When microbiological results were added to clinical criteria, adequate diagnoses originating from microbiological results which might have corrected false positive and false negative clinical judgements (n = 5) were countered by a similar proportion of inadequate diagnoses (n = 6). Clinical criteria had reasonable diagnostic values. CPIS was not superior to conventional clinical criteria. Non-invasive and invasive sampling techniques had diagnostic values comparable to clinical criteria. An algorithm guiding antibiotic treatment exclusively by microbiological results does not increase the overall diagnostic accuracy and carries the risk of undertreatment.

High-Grading Lunar Samples

NASA Technical Reports Server (NTRS)

Allen, Carlton; Sellar, Glenn; Nunez, Jorge; Mosie, Andrea; Schwarz, Carol; Parker, Terry; Winterhalter, Daniel; Farmer, Jack

2009-01-01

Astronauts on long-duration lunar missions will need the capability to high-grade their samples to select the highest value samples for transport to Earth and to leave others on the Moon. We are supporting studies to define the necessary and sufficient measurements and techniques for high-grading samples at a lunar outpost. A glovebox, dedicated to testing instruments and techniques for high-grading samples, is in operation at the JSC Lunar Experiment Laboratory. A reference suite of lunar rocks and soils, spanning the full compositional range found in the Apollo collection, is available for testing in this laboratory. Thin sections of these samples are available for direct comparison. The Lunar Sample Compendium, on-line at http://www-curator.jsc.nasa.gov/lunar/compendium.cfm, summarizes previous analyses of these samples. The laboratory, sample suite, and Compendium are available to the lunar research and exploration community. In the first test of possible instruments for lunar sample high-grading, we imaged 18 lunar rocks and four soils from the reference suite using the Multispectral Microscopic Imager (MMI) developed by Arizona State University and JPL (see Farmer et. al. abstract). The MMI is a fixed-focus digital imaging system with a resolution of 62.5 microns/pixel, a field size of 40 x 32 mm, and a depth-of-field of approximately 5 mm. Samples are illuminated sequentially by 21 light emitting diodes in discrete wavelengths spanning the visible to shortwave infrared. Measurements of reflectance standards and background allow calibration to absolute reflectance. ENVI-based software is used to produce spectra for specific minerals as well as multi-spectral images of rock textures.

Efficient electromagnetic source imaging with adaptive standardized LORETA/FOCUSS.

PubMed

Schimpf, Paul H; Liu, Hesheng; Ramon, Ceon; Haueisen, Jens

2005-05-01

Functional brain imaging and source localization based on the scalp's potential field require a solution to an ill-posed inverse problem with many solutions. This makes it necessary to incorporate a priori knowledge in order to select a particular solution. A computational challenge for some subject-specific head models is that many inverse algorithms require a comprehensive sampling of the candidate source space at the desired resolution. In this study, we present an algorithm that can accurately reconstruct details of localized source activity from a sparse sampling of the candidate source space. Forward computations are minimized through an adaptive procedure that increases source resolution as the spatial extent is reduced. With this algorithm, we were able to compute inverses using only 6% to 11% of the full resolution lead-field, with a localization accuracy that was not significantly different than an exhaustive search through a fully-sampled source space. The technique is, therefore, applicable for use with anatomically-realistic, subject-specific forward models for applications with spatially concentrated source activity.

Detection of Legionella pneumophila by PCR-ELISA method in industrial cooling tower water.

PubMed

Soheili, Majid; Nejadmoghaddam, Mohammad Reza; Babashamsi, Mohammad; Ghasemi, Jamileh; Jeddi Tehrani, Mahmood

2007-11-15

Water supply and Cooling Tower Water (CTW) are among the most common sources of Legionella pneumophila (LP) contamination. A nonradio active method is described to detect LP in industrial CTW samples. DNA was purified and amplified by nested -PCR with amplimers specific for the 16s rRNA gene of LP. The 5' end biotinylated oligomer probe was immobilized on sterptavidin B coated microtiter plates. The nested-PCR product was labeled with digoxigenin and then hybridized with 5'-biotinylated probes. The amplification products were detected by using proxidase-labled anti dioxygenin antibody in a colorimetric reaction. The assay detected LP present in 1 L of 5 CTW samples examined. All of the samples were Legionella positive in both culture and PCR-ELISA methods. The PCR-ELISA assay appears to exhibit high specificity and is a more rapid technique in comparison with bacterial culture method. Thus could prove suitable for use in the routine examination of industrial CTW contamination.

In-house PCR with DNA extracted directly from positive slides to confirm or exclude the diagnosis of tuberculosis: focus on biosafety.

PubMed

de Almeida, Isabela Neves; Aleixo, Agdemir Valéria; Carvalho, Wânia da Silva; de Miranda, Silvana Spindola

2015-01-01

The possibility to obtain DNA from smears is a valuable alternative to remedy the lack of samples when they are totally used for bacilloscopy; this technique solves the biosafety problem related to a possible accident with the transportation of flasks containing potentially transmissible clinical samples. Hence, the purpose of this study was to utilize the insertion sequence IS6110 for amplification of DNA from a smear-positive sample for tuberculosis (TB) diagnosis. Among the 52 positive bacilloscopies, sensitivity, specificity, positive predictive value and negative predictive value were 52.3%, 100%, 100% and 89.7%, respectively whereas accuracy was 90.7%. The IS6110-based PCR for TB diagnosis developed in DNA extracted from a positive smear is a fast, simple, specific, and safe method. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories

PubMed Central

Akimoto, Chizuru; Volk, Alexander E; van Blitterswijk, Marka; Van den Broeck, Marleen; Leblond, Claire S; Lumbroso, Serge; Camu, William; Neitzel, Birgit; Onodera, Osamu; van Rheenen, Wouter; Pinto, Susana; Weber, Markus; Smith, Bradley; Proven, Melanie; Talbot, Kevin; Keagle, Pamela; Chesi, Alessandra; Ratti, Antonia; van der Zee, Julie; Alstermark, Helena; Birve, Anna; Calini, Daniela; Nordin, Angelica; Tradowsky, Daniela C; Just, Walter; Daoud, Hussein; Angerbauer, Sabrina; DeJesus-Hernandez, Mariely; Konno, Takuya; Lloyd-Jani, Anjali; de Carvalho, Mamede; Mouzat, Kevin; Landers, John E; Veldink, Jan H; Silani, Vincenzo; Gitler, Aaron D; Shaw, Christopher E; Rouleau, Guy A; van den Berg, Leonard H; Van Broeckhoven, Christine; Rademakers, Rosa; Andersen, Peter M; Kubisch, Christian

2014-01-01

Background The GGGGCC-repeat expansion in C9orf72 is the most frequent mutation found in patients with amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Most of the studies on C9orf72 have relied on repeat-primed PCR (RP-PCR) methods for detection of the expansions. To investigate the inherent limitations of this technique, we compared methods and results of 14 laboratories. Methods The 14 laboratories genotyped DNA from 78 individuals (diagnosed with ALS or FTD) in a blinded fashion. Eleven laboratories used a combination of amplicon-length analysis and RP-PCR, whereas three laboratories used RP-PCR alone; Southern blotting techniques were used as a reference. Results Using PCR-based techniques, 5 of the 14 laboratories got results in full accordance with the Southern blotting results. Only 50 of the 78 DNA samples got the same genotype result in all 14 laboratories. There was a high degree of false positive and false negative results, and at least one sample could not be genotyped at all in 9 of the 14 laboratories. The mean sensitivity of a combination of amplicon-length analysis and RP-PCR was 95.0% (73.9–100%), and the mean specificity was 98.0% (87.5–100%). Overall, a sensitivity and specificity of more than 95% was observed in only seven laboratories. Conclusions Because of the wide range seen in genotyping results, we recommend using a combination of amplicon-length analysis and RP-PCR as a minimum in a research setting. We propose that Southern blotting techniques should be the gold standard, and be made obligatory in a clinical diagnostic setting. PMID:24706941

[The diagnosis of malaria by the thick film and the QBC: a comparative study of both technics].

PubMed

Cabezos, J; Bada, J L

1993-06-12

The diagnosis of paludism is important because of the severity of the clinical picture caused by Plasmodium falciparum, the increasing number of travellers to endemic zones and the emigration from these zones. A comparative study of the QBC techniques (staining with acridin orange and observation with ultraviolet light) and the thick film with Giemsa staining was carried out. The QBC and thick film were performed parallelly for 17 months in a total of 623 samples pertaining to subjects from endemic zones of paludism (emigrants, immigrants and travellers). Of the 623 samples studied 49 were positive for paludism by both techniques. Ten were positive with only the thick film and six were positive only with QBC. The sensitivity of QBC versus thick film was 83% and specificity 98.9%. The time used to determine diagnosis with the QBC technique ranged from 6 to 12 minutes from withdrawal of the sample, while with the thick film the time spent was more than 2 hours. The cases positive by thick film and negative with QBC corresponded to patients with very low parasitation. The intensity of parasitation was difficult to determine quantitatively by QBC. Although the QBC technique has the advantage of speed it is inexact with respect to the quantification of parasitemia. Moreover, it is less sensitive than the thick film in patient with very low parasitations and cannot thus substitute the thick film.

Single-molecule fluorescence microscopy review: shedding new light on old problems

PubMed Central

Shashkova, Sviatlana

2017-01-01

Fluorescence microscopy is an invaluable tool in the biosciences, a genuine workhorse technique offering exceptional contrast in conjunction with high specificity of labelling with relatively minimal perturbation to biological samples compared with many competing biophysical techniques. Improvements in detector and dye technologies coupled to advances in image analysis methods have fuelled recent development towards single-molecule fluorescence microscopy, which can utilize light microscopy tools to enable the faithful detection and analysis of single fluorescent molecules used as reporter tags in biological samples. For example, the discovery of GFP, initiating the so-called ‘green revolution’, has pushed experimental tools in the biosciences to a completely new level of functional imaging of living samples, culminating in single fluorescent protein molecule detection. Today, fluorescence microscopy is an indispensable tool in single-molecule investigations, providing a high signal-to-noise ratio for visualization while still retaining the key features in the physiological context of native biological systems. In this review, we discuss some of the recent discoveries in the life sciences which have been enabled using single-molecule fluorescence microscopy, paying particular attention to the so-called ‘super-resolution’ fluorescence microscopy techniques in live cells, which are at the cutting-edge of these methods. In particular, how these tools can reveal new insights into long-standing puzzles in biology: old problems, which have been impossible to tackle using other more traditional tools until the emergence of new single-molecule fluorescence microscopy techniques. PMID:28694303

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

PubMed

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

2009-01-01

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

Nanoscale chemical imaging by photoinduced force microscopy

PubMed Central

Nowak, Derek; Morrison, William; Wickramasinghe, H. Kumar; Jahng, Junghoon; Potma, Eric; Wan, Lei; Ruiz, Ricardo; Albrecht, Thomas R.; Schmidt, Kristin; Frommer, Jane; Sanders, Daniel P.; Park, Sung

2016-01-01

Correlating spatial chemical information with the morphology of closely packed nanostructures remains a challenge for the scientific community. For example, supramolecular self-assembly, which provides a powerful and low-cost way to create nanoscale patterns and engineered nanostructures, is not easily interrogated in real space via existing nondestructive techniques based on optics or electrons. A novel scanning probe technique called infrared photoinduced force microscopy (IR PiFM) directly measures the photoinduced polarizability of the sample in the near field by detecting the time-integrated force between the tip and the sample. By imaging at multiple IR wavelengths corresponding to absorption peaks of different chemical species, PiFM has demonstrated the ability to spatially map nm-scale patterns of the individual chemical components of two different types of self-assembled block copolymer films. With chemical-specific nanometer-scale imaging, PiFM provides a powerful new analytical method for deepening our understanding of nanomaterials. PMID:27051870

Combination of complementary data mining methods for geographical characterization of extra virgin olive oils based on mineral composition.

PubMed

Sayago, Ana; González-Domínguez, Raúl; Beltrán, Rafael; Fernández-Recamales, Ángeles

2018-09-30

This work explores the potential of multi-element fingerprinting in combination with advanced data mining strategies to assess the geographical origin of extra virgin olive oil samples. For this purpose, the concentrations of 55 elements were determined in 125 oil samples from multiple Spanish geographic areas. Several unsupervised and supervised multivariate statistical techniques were used to build classification models and investigate the relationship between mineral composition of olive oils and their provenance. Results showed that Spanish extra virgin olive oils exhibit characteristic element profiles, which can be differentiated on the basis of their origin in accordance with three geographical areas: Atlantic coast (Huelva province), Mediterranean coast and inland regions. Furthermore, statistical modelling yielded high sensitivity and specificity, principally when random forest and support vector machines were employed, thus demonstrating the utility of these techniques in food traceability and authenticity research. Copyright © 2018 Elsevier Ltd. All rights reserved.

Halal authenticity issues in meat and meat products.

PubMed

Nakyinsige, Khadijah; Man, Yaakob Bin Che; Sazili, Awis Qurni

2012-07-01

In the recent years, Muslims have become increasingly concerned about the meat they eat. Proper product description is very crucial for consumers to make informed choices and to ensure fair trade, particularly in the ever growing halal food market. Globally, Muslim consumers are concerned about a number of issues concerning meat and meat products such as pork substitution, undeclared blood plasma, use of prohibited ingredients, pork intestine casings and non-halal methods of slaughter. Analytical techniques which are appropriate and specific have been developed to deal with particular issues. The most suitable technique for any particular sample is often determined by the nature of the sample itself. This paper sets out to identify what makes meat halal, highlight the halal authenticity issues that occur in meat and meat products and provide an overview of the possible analytical methods for halal authentication of meat and meat products. Copyright © 2012 Elsevier Ltd. All rights reserved.

Design of Measurement Apparatus for Electromagnetic Shielding Effectiveness Using Flanged Double Ridged Waveguide

NASA Astrophysics Data System (ADS)

Kwon, Jong Hwa; Choi, Jae Ick; Yook, Jong Gwan

In this paper, we design and manufacture a flanged double ridged waveguide with a tapered section as a sample holder for measuring the electromagnetic shielding effectiveness (SE) of planar material in broadband frequency ranges up to 10GHz. The proposed technique overcomes the limitations of the conventional ASTM D4935 test method at high frequencies. The simulation results for the designed sample holders agree well with the fabricated ones in consideration of the design specification of S11 < -20dB within the frequency range of 1-10GHz. To verify the proposed measurement apparatus, the measured SE data of the commercial shielding materials from 1 to 10GHz were indirectly compared with those obtained from the ASTM D4935 from 30MHz to 1GHz. We observed that the SE data obtained by using both experimental techniques agree with each other.

Metal and proton adsorption capacities of natural and cloned Sphagnum mosses.

PubMed

Gonzalez, Aridane G; Pokrovsky, Oleg S; Beike, Anna K; Reski, Ralf; Di Palma, Anna; Adamo, Paola; Giordano, Simonetta; Angel Fernandez, J

2016-01-01

Terrestrial mosses are commonly used as bioindicators of atmospheric pollution. However, there is a lack of standardization of the biomonitoring preparation technique and the efficiency of metal adsorption by various moss species is poorly known. This is especially true for in vitro-cultivated moss clones, which are promising candidates for a standardized moss-bag technique. We studied the adsorption of copper and zinc on naturally grown Sphagnum peat moss in comparison with in vitro-cultivated Sphagnum palustre samples in order to provide their physico-chemical characterization and to test the possibility of using cloned peat mosses as bioindicators within the protocol of moss-bag technique. We demonstrate that in vitro-grown clones of S. palustre exhibit acid-base properties similar to those of naturally grown Sphagnum samples, whereas the zinc adsorption capacity of the clones is approx. twice higher than that of the samples from the field. At the same time, the field samples adsorbed 30-50% higher amount of Cu(2+) compared to that of the clones. This contrast may be related to fine differences in the bulk chemical composition, specific surface area, morphological features, type and abundance of binding sites at the cell surfaces and in the aqueous solution of natural and cloned Sphagnum. The clones exhibited much lower concentration of most metal pollutants in their tissues relative to the natural samples thus making the former better indicators of low metal loading. Overall, in vitro-produced clones of S. palustre can be considered as an adequate, environmentally benign substitution for protected natural Sphagnum sp. samples to be used in moss-bags for atmospheric monitoring. Copyright © 2015 Elsevier Inc. All rights reserved.

Separation techniques for the clean-up of radioactive mixed waste for ICP-AES/ICP-MS analysis

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

Swafford, A.M.; Keller, J.M.

1993-03-17

Two separation techniques were investigated for the clean-up of typical radioactive mixed waste samples requiring elemental analysis by Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) or Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). These measurements frequently involve regulatory or compliance criteria which include the determination of elements on the EPA Target Analyte List (TAL). These samples usually consist of both an aqueous phase and a solid phase which is mostly an inorganic sludge. Frequently, samples taken from the waste tanks contain high levels of uranium and thorium which can cause spectral interferences in ICP-AES or ICP-MS analysis. The removal of these interferences ismore » necessary to determine the presence of the EPA TAL elements in the sample. Two clean-up methods were studied on simulated aqueous waste samples containing the EPA TAL elements. The first method studied was a classical procedure based upon liquid-liquid extraction using tri-n- octylphosphine oxide (TOPO) dissolved in cyclohexane. The second method investigated was based on more recently developed techniques using extraction chromatography; specifically the use of a commercially available Eichrom TRU[center dot]Spec[trademark] column. Literature on these two methods indicates the efficient removal of uranium and thorium from properly prepared samples and provides considerable qualitative information on the extraction behavior of many other elements. However, there is a lack of quantitative data on the extraction behavior of elements on the EPA Target Analyte List. Experimental studies on these two methods consisted of determining whether any of the analytes were extracted by these methods and the recoveries obtained. Both methods produced similar results; the EPA target analytes were only slightly or not extracted. Advantages and disadvantages of each method were evaluated and found to be comparable.« less

Electrical field-induced extraction and separation techniques: promising trends in analytical chemistry--a review.

PubMed

Yamini, Yadollah; Seidi, Shahram; Rezazadeh, Maryam

2014-03-03

Sample preparation is an important issue in analytical chemistry, and is often a bottleneck in chemical analysis. So, the major incentive for the recent research has been to attain faster, simpler, less expensive, and more environmentally friendly sample preparation methods. The use of auxiliary energies, such as heat, ultrasound, and microwave, is one of the strategies that have been employed in sample preparation to reach the above purposes. Application of electrical driving force is the current state-of-the-art, which presents new possibilities for simplifying and shortening the sample preparation process as well as enhancing its selectivity. The electrical driving force has scarcely been utilized in comparison with other auxiliary energies. In this review, the different roles of electrical driving force (as a powerful auxiliary energy) in various extraction techniques, including liquid-, solid-, and membrane-based methods, have been taken into consideration. Also, the references have been made available, relevant to the developments in separation techniques and Lab-on-a-Chip (LOC) systems. All aspects of electrical driving force in extraction and separation methods are too specific to be treated in this contribution. However, the main aim of this review is to provide a brief knowledge about the different fields of analytical chemistry, with an emphasis on the latest efforts put into the electrically assisted membrane-based sample preparation systems. The advantages and disadvantages of these approaches as well as the new achievements in these areas have been discussed, which might be helpful for further progress in the future. Copyright © 2013 Elsevier B.V. All rights reserved.

Laboratory blood analysis in Strigiformes-Part I: hematologic reference intervals and agreement between manual blood cell counting techniques.

PubMed

Ammersbach, Mélanie; Beaufrère, Hugues; Gionet Rollick, Annick; Tully, Thomas

2015-03-01

While hematologic reference intervals (RI) are available for multiple raptorial species of the order Accipitriformes and Falconiformes, there is a lack of valuable hematologic information in Strigiformes that can be used for diagnostic and health monitoring purposes. The objective was to report RI in Strigiformes for hematologic variables and to assess agreement between manual cell counting techniques. A multi-center prospective study was designed to assess hematologic RI and blood cell morphology in owl species. Samples were collected from individuals representing 13 Strigiformes species, including Great Horned Owl, Snowy Owl, Eurasian Eagle Owl, Barred Owl, Great Gray Owl, Ural Owl, Northern Saw-Whet Owls, Northern Hawk Owl, Spectacled Owl, Barn Owl, Eastern Screech Owl, Long-Eared Owl, and Short-Eared Owl. Red blood cell count was determined manually using a hemocytometer. White blood cell count was determined using 3 manual counting techniques: (1) phloxine B technique, (2) Natt and Herrick technique, and (3) estimation from the smear. Differential counts and blood cell morphology were determined on smears. Reference intervals were determined and agreement between methods was calculated. Important species-specific differences were observed in blood cell counts and granulocyte morphology. Differences in WBC count between species did not appear to be predictable based on phylogenetic relationships. Overall, most boreal owl species exhibited a lower WBC count than other species. Important disagreements were found between different manual WBC counting techniques. Disagreements observed between manual counting techniques suggest that technique-specific RI should be used in Strigiformes. © 2015 American Society for Veterinary Clinical Pathology.

Rapid Radiochemical Methods for Asphalt Paving Material ...

EPA Pesticide Factsheets

Technical Brief Validated rapid radiochemical methods for alpha and beta emitters in solid matrices that are commonly encountered in urban environments were previously unavailable for public use by responding laboratories. A lack of tested rapid methods would delay the quick determination of contamination levels and the assessment of acceptable site-specific exposure levels. Of special concern are matrices with rough and porous surfaces, which allow the movement of radioactive material deep into the building material making it difficult to detect. This research focuses on methods that address preparation, radiochemical separation, and analysis of asphalt paving materials and asphalt roofing shingles. These matrices, common to outdoor environments, challenge the capability and capacity of very experienced radiochemistry laboratories. Generally, routine sample preparation and dissolution techniques produce liquid samples (representative of the original sample material) that can be processed using available radiochemical methods. The asphalt materials are especially difficult because they do not readily lend themselves to these routine sample preparation and dissolution techniques. The HSRP and ORIA coordinate radiological reference laboratory priorities and activities in conjunction with HSRP’s Partner Process. As part of the collaboration, the HSRP worked with ORIA to publish rapid radioanalytical methods for selected radionuclides in building material matrice

UV EFFECTS IN TOOTH ENAMEL AND THEIR POSSIBLE APPLICATION IN EPR DOSIMETRY WITH FRONT TEETH

PubMed Central

Sholom, S.; Desrosiers, M.; Chumak, V.; Luckyanov, N.; Simon, S.L.; Bouville, A.

2009-01-01

The effects of ultraviolet (UV) radiation on ionizing radiation biodosimetry were studied in human tooth enamel samples using the technique of electron paramagnetic resonance (EPR) in X-band. For samples in the form of grains, UV-specific EPR spectra were spectrally distinct from that produced by exposure to gamma radiation. From larger enamel samples, the UV penetration depth was determined to be in the 60–120 μm range. The difference in EPR spectra from UV exposure and from exposure to gamma radiation samples was found to be a useful marker of UV equivalent dose (defined as the apparent contribution to the gamma dose in mGy that results from UV radiation absorption) in tooth enamel. This concept was preliminarily tested on front teeth from inhabitants of the region of the Semipalatinsk Nuclear Test Site (Kazakhstan) who might have received some exposure to gamma radiation from the nuclear tests conducted there as well as from normal UV radiation in sunlight. The technique developed here to quantify and subtract the UV contribution to the measured tooth is currently limited to cumulative dose measurements with a component of UV equivalent dose equal to or greater than 300 mGy. PMID:20065706

Modulated Raman spectroscopy for enhanced identification of bladder tumor cells in urine samples.

PubMed

Canetta, Elisabetta; Mazilu, Michael; De Luca, Anna Chiara; Carruthers, Antonia E; Dholakia, Kishan; Neilson, Sam; Sargeant, Harry; Briscoe, Tina; Herrington, C Simon; Riches, Andrew C

2011-03-01

Standard Raman spectroscopy (SRS) is a noninvasive technique that is used in the biomedical field to discriminate between normal and cancer cells. However, the presence of a strong fluorescence background detracts from the use of SRS in real-time clinical applications. Recently, we have reported a novel modulated Raman spectroscopy (MRS) technique to extract the Raman spectra from the background. In this paper, we present the first application of MRS to the identification of human urothelial cells (SV-HUC-1) and bladder cancer cells (MGH) in urine samples. These results are compared to those obtained by SRS. Classification using the principal component analysis clearly shows that MRS allows discrimination between Raman spectra of SV-HUC-1 and MGH cells with high sensitivity (98%) and specificity (95%). MRS is also used to distinguish between SV-HUC-1 and MGH cells after exposure to urine for up to 6 h. We observe a marked change in the MRS of SV-HUC-1 and MGH cells with time in urine, indicating that the conditions of sample collection will be important for the application of this methodology to clinical urine samples.

Laser-induced breakdown spectroscopy for detection of heavy metals in environmental samples

NASA Astrophysics Data System (ADS)

Wisbrun, Richard W.; Schechter, Israel; Niessner, Reinhard; Schroeder, Hartmut

1993-03-01

The application of LIBS technology as a sensor for heavy metals in solid environmental samples has been studied. This specific application introduces some new problems in the LIBS analysis. Some of them are related to the particular distribution of contaminants in the grained samples. Other problems are related to mechanical properties of the samples and to general matrix effects, like the water and organic fibers content of the sample. An attempt has been made to optimize the experimental set-up for the various involved parameters. The understanding of these factors has enabled the adjustment of the technique to the substrates of interest. The special importance of the grain size and of the laser-induced aerosol production is pointed out. Calibration plots for the analysis of heavy metals in diverse sand and soil samples have been carried out. The detection limits are shown to be usually below the recent regulation restricted concentrations.

Measurements of 55Fe activity in activated steel samples with GEMPix

NASA Astrophysics Data System (ADS)

Curioni, A.; Dinar, N.; La Torre, F. P.; Leidner, J.; Murtas, F.; Puddu, S.; Silari, M.

2017-03-01

In this paper we present a novel method, based on the recently developed GEMPix detector, to measure the 55Fe content in samples of metallic material activated during operation of CERN accelerators and experimental facilities. The GEMPix, a gas detector with highly pixelated read-out, has been obtained by coupling a triple Gas Electron Multiplier (GEM) to a quad Timepix ASIC. Sample preparation, measurements performed on 45 samples and data analysis are described. The calibration factor (counts per second per unit specific activity) has been obtained via measurements of the 55Fe activity determined by radiochemical analysis of the same samples. Detection limit and sensitivity to the current Swiss exemption limit are calculated. Comparison with radiochemical analysis shows inconsistency for the sensitivity for only two samples, most likely due to underestimated uncertainties of the GEMPix analysis. An operative test phase of this technique is already planned at CERN.

Optimal design in pediatric pharmacokinetic and pharmacodynamic clinical studies.

PubMed

Roberts, Jessica K; Stockmann, Chris; Balch, Alfred; Yu, Tian; Ward, Robert M; Spigarelli, Michael G; Sherwin, Catherine M T

2015-03-01

It is not trivial to conduct clinical trials with pediatric participants. Ethical, logistical, and financial considerations add to the complexity of pediatric studies. Optimal design theory allows investigators the opportunity to apply mathematical optimization algorithms to define how to structure their data collection to answer focused research questions. These techniques can be used to determine an optimal sample size, optimal sample times, and the number of samples required for pharmacokinetic and pharmacodynamic studies. The aim of this review is to demonstrate how to determine optimal sample size, optimal sample times, and the number of samples required from each patient by presenting specific examples using optimal design tools. Additionally, this review aims to discuss the relative usefulness of sparse vs rich data. This review is intended to educate the clinician, as well as the basic research scientist, whom plan on conducting a pharmacokinetic/pharmacodynamic clinical trial in pediatric patients. © 2015 John Wiley & Sons Ltd.

Rapid and sensitive detection of human astrovirus in water samples by loop-mediated isothermal amplification with hydroxynaphthol blue dye.

PubMed

Yang, Bo-Yun; Liu, Xiao-Lu; Wei, Yu-Mei; Wang, Jing-Qi; He, Xiao-Qing; Jin, Yi; Wang, Zi-Jian

2014-02-14

The aim of this paper was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid, sensitive and inexpensive detection of astrovirus. The detection limit of LAMP using in vitro RNA transcripts was 3.6 × 10 copies·μL⁻¹, which is as sensitive as the presently used PCR assays. However, the LAMP products could be identified as different colors with the naked eye following staining with hydroxynaphthol blue dye (HNB). No cross-reactivity with other gastroenteric viruses (rotavirus and norovirus) was observed, indicating the relatively high specificity of LAMP. The RT-LAMP method with HNB was used to effectively detect astrovirus in reclaimed water samples. The LAMP technique described in this study is a cheap, sensitive, specific and rapid method for the detection of astrovirus. The RT-LAMP method can be simply applied for the specific detection of astrovirus and has the potential to be utilized in the field as a screening test.

The vector homology problem in diagnostic nucleic acid hybridization of clinical specimens.

PubMed Central

Ambinder, R F; Charache, P; Staal, S; Wright, P; Forman, M; Hayward, S D; Hayward, G S

1986-01-01

Nucleic acid hybridization techniques using cloned probes are finding application in assays of clinical specimens in research and diagnostic laboratories. The probes that we and others have used are recombinant plasmids composed of viral inserts and bacterial plasmid vectors such as pBR322. We suspected that there was material homologous to pBR322 present in many clinical samples. because hybridization occurred in samples which lacked evidence of virus by other techniques. If the presence of this vector-homologous material was unrecognized, hybridization in the test sample might erroneously be interpreted as indicating the presence of viral sequences. In this paper we demonstrate specific hybridization of labeled pBR322 DNA with DNA from various clinical samples. Evidence is presented that nonspecific probe trapping could not account for this phenomenon. In mixing experiments, it is shown that contamination of clinical samples with bacteria would explain such a result. Approaches tested to circumvent this problem included the use of isolated insert probes, alternate cloning vectors, and cold competitor pBR322 DNA in prehybridization and hybridization mixes. None proved entirely satisfactory. We therefore emphasize that it is essential that all hybridization detection systems use a control probe of the vector alone in order to demonstrate the absence of material with vector homology in the specimen tested. Images PMID:3013928

Measurement of plutonium isotope ratios in nuclear fuel samples by HPLC-MC-ICP-MS

NASA Astrophysics Data System (ADS)

Günther-Leopold, I.; Waldis, J. Kobler; Wernli, B.; Kopajtic, Z.

2005-04-01

Radioactive isotopes are traditionally quantified by means of radioactivity counting techniques ([alpha], [beta], [gamma]). However, these methods often require extensive matrix separation and sample purification before the identification of specific isotopes and their relative abundance is possible as it is necessary in the frame of post-irradiation examinations on nuclear fuel samples. The technique of multicollector inductively coupled plasma mass spectrometry (MC-ICP-MS) is attracting much attention because it permits the precise measurement of the isotope compositions for a wide range of elements combined with excellent limits of detection due to high ionization efficiencies. The present paper describes one of the first applications of an online high-performance liquid chromatographic separation system coupled to a MC-ICP-MS in order to overcome isobaric interferences for the determination of the plutonium isotope composition and concentrations in irradiated nuclear fuels. The described chromatographic separation is sufficient to prevent any isobaric interference between 238Pu present at trace concentrations and 238U present as the main component of the fuel samples. The external reproducibility of the uncorrected plutonium isotope ratios was determined to be between 0.04 and 0.2% (2 s) resulting in a precision in the [per mille sign] range for the isotopic vectors of the irradiated fuel samples.