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Sample records for high-resolution melt polymerase

  1. Automated Microfluidic Platform for Serial Polymerase Chain Reaction and High-Resolution Melting Analysis.

    Science.gov (United States)

    Cao, Weidong; Bean, Brian; Corey, Scott; Coursey, Johnathan S; Hasson, Kenton C; Inoue, Hiroshi; Isano, Taisuke; Kanderian, Sami; Lane, Ben; Liang, Hongye; Murphy, Brian; Owen, Greg; Shinoda, Nobuhiko; Zeng, Shulin; Knight, Ivor T

    2016-06-01

    We report the development of an automated genetic analyzer for human sample testing based on microfluidic rapid polymerase chain reaction (PCR) with high-resolution melting analysis (HRMA). The integrated DNA microfluidic cartridge was used on a platform designed with a robotic pipettor system that works by sequentially picking up different test solutions from a 384-well plate, mixing them in the tips, and delivering mixed fluids to the DNA cartridge. A novel image feedback flow control system based on a Canon 5D Mark II digital camera was developed for controlling fluid movement through a complex microfluidic branching network without the use of valves. The same camera was used for measuring the high-resolution melt curve of DNA amplicons that were generated in the microfluidic chip. Owing to fast heating and cooling as well as sensitive temperature measurement in the microfluidic channels, the time frame for PCR and HRMA was dramatically reduced from hours to minutes. Preliminary testing results demonstrated that rapid serial PCR and HRMA are possible while still achieving high data quality that is suitable for human sample testing. © 2015 Society for Laboratory Automation and Screening.

  2. Duchenne muscular dystrophy: High-resolution melting curve ...

    African Journals Online (AJOL)

    Duchenne muscular dystrophy: High-resolution melting curve analysis as an affordable diagnostic mutation scanning tool in a South African cohort. ... Genetic screening for D/BMD in South Africa currently includes multiple ligase-dependent probe amplification (MLPA) for exonic deletions and duplications and linkage ...

  3. Methylation-Sensitive High Resolution Melting (MS-HRM).

    Science.gov (United States)

    Hussmann, Dianna; Hansen, Lise Lotte

    2018-01-01

    Methylation-Sensitive High Resolution Melting (MS-HRM) is an in-tube, PCR-based method to detect methylation levels at specific loci of interest. A unique primer design facilitates a high sensitivity of the assays enabling detection of down to 0.1-1% methylated alleles in an unmethylated background.Primers for MS-HRM assays are designed to be complementary to the methylated allele, and a specific annealing temperature enables these primers to anneal both to the methylated and the unmethylated alleles thereby increasing the sensitivity of the assays. Bisulfite treatment of the DNA prior to performing MS-HRM ensures a different base composition between methylated and unmethylated DNA, which is used to separate the resulting amplicons by high resolution melting.The high sensitivity of MS-HRM has proven useful for detecting cancer biomarkers in a noninvasive manner in urine from bladder cancer patients, in stool from colorectal cancer patients, and in buccal mucosa from breast cancer patients. MS-HRM is a fast method to diagnose imprinted diseases and to clinically validate results from whole-epigenome studies. The ability to detect few copies of methylated DNA makes MS-HRM a key player in the quest for establishing links between environmental exposure, epigenetic changes, and disease.

  4. High Resolution Melting (HRM) applied to wine authenticity.

    Science.gov (United States)

    Pereira, Leonor; Gomes, Sónia; Castro, Cláudia; Eiras-Dias, José Eduardo; Brazão, João; Graça, António; Fernandes, José R; Martins-Lopes, Paula

    2017-02-01

    Wine authenticity methods are in increasing demand mainly in Denomination of Origin designations. The DNA-based methodologies are a reliable means of tracking food/wine varietal composition. The main aim of this work was the study of High Resolution Melting (HRM) application as a screening method for must and wine authenticity. Three sample types (leaf, must and wine) were used to validate the three developed HRM assays (Vv1-705bp; Vv2-375bp; and Vv3-119bp). The Vv1 HRM assay was only successful when applied to leaf and must samples. The Vv2 HRM assay successfully amplified all sample types, allowing genotype discrimination based on melting temperature values. The smallest amplicon, Vv3, produced a coincident melting curve shape in all sample types (leaf and wine) with corresponding genotypes. This study presents sensitive, rapid and efficient HRM assays applied for the first time to wine samples suitable for wine authenticity purposes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Identification of a new genetic marker in Mycoplasma synoviae vaccine strain MS-H and development of a strategy using polymerase chain reaction and high-resolution melting curve analysis for differentiating MS-H from field strains.

    Science.gov (United States)

    Zhu, Ling; Konsak, Barbara M; Olaogun, Olusola M; Agnew-Crumptona, Rebecca; Kanci, Anna; Marenda, Marc S; Browning, Glenn F; Noormohammadi, Amir H

    2017-10-01

    Mycoplasma synoviae (MS) is an economically important avian pathogen worldwide, causing subclinical respiratory tract infection and infectious synovitis in chickens and turkeys. A temperature-sensitive (ts + ) live attenuated vaccine MS-H, derived from the Australian field strain 86079/7NS, is now widely used in many countries to control the disease induced by MS. Differentiation of MS-H vaccine from field strains is crucial for monitoring vaccination programs in commercial poultry. Comparison of genomic sequences of MS-H and its parent strain revealed an adenine deletion at nucleotide position 468 of the MS-H oppF-1 gene. This mutation was shown to be unique to MS-H in further comparative analyses of oppF-1 genes of MS-H re-isolates and field strains from Australia and other countries. Based on this single nucleotide, a combination of nested PCR and high-resolution melting (HRM) curve analysis was used to evaluate its potential for use in differentiation of MS-H from field strains. The mean genotype confidence percentages of 99.27 and 48.20 for MS-H and field strains, respectively, demonstrated the high discriminative power of the newly developed assay (oppF PCR-HRM). A set of 13 tracheal swab samples collected from MS-H vaccinated specific pathogen free birds and commercial chicken flocks infected with MS were tested using the oppF PCR-HRM test and results were totally consistent with those obtained using vlhA genotyping. The nested-PCR HRM method established in this study proved to be a rapid, simple and cost effective tool for discriminating the MS-H vaccine strain from Australian and international strains in pure cultures and on tracheal swabs. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. High resolution melting (HRM) analysis of DNA--its role and potential in food analysis.

    Science.gov (United States)

    Druml, Barbara; Cichna-Markl, Margit

    2014-09-01

    DNA based methods play an increasing role in food safety control and food adulteration detection. Recent papers show that high resolution melting (HRM) analysis is an interesting approach. It involves amplification of the target of interest in the presence of a saturation dye by the polymerase chain reaction (PCR) and subsequent melting of the amplicons by gradually increasing the temperature. Since the melting profile depends on the GC content, length, sequence and strand complementarity of the product, HRM analysis is highly suitable for the detection of single-base variants and small insertions or deletions. The review gives an introduction into HRM analysis, covers important aspects in the development of an HRM analysis method and describes how HRM data are analysed and interpreted. Then we discuss the potential of HRM analysis based methods in food analysis, i.e. for the identification of closely related species and cultivars and the identification of pathogenic microorganisms. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Genotyping of Listeria monocytogenes isolates from poultry carcasses using high resolution melting (HRM) analysis.

    Science.gov (United States)

    Sakaridis, Ioannis; Ganopoulos, Ioannis; Madesis, Panagiotis; Tsaftaris, Athanasios; Argiriou, Anagnostis

    2014-01-02

    An outbreak situation of human listeriosis requires a fast and accurate protocol for typing Listeria monocytogenes . Existing techniques are either characterized by low discriminatory power or are laborious and require several days to give a final result. Polymerase chain reaction (PCR) coupled with high resolution melting (HRM) analysis was investigated in this study as an alternative tool for a rapid and precise genotyping of L. monocytogenes isolates. Fifty-five isolates of L. monocytogenes isolated from poultry carcasses and the environment of four slaughterhouses were typed by HRM analysis using two specific markers, internalin B and ssrA genes. The analysis of genotype confidence percentage of L. monocytogenes isolates produced by HRM analysis generated dendrograms with two major groups and several subgroups. Furthermore, the analysis of the HRM curves revealed that all L. monocytogenes isolates could easily be distinguished. In conclusion, HRM was proven to be a fast and powerful tool for genotyping isolates of L. monocytogenes .

  8. High Resolution Melting Analysis for fast and cheap polymorphism screening of marine populations

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Anne-Leila Meistertzheim, Isabelle Calves, Sébastien Artigaud, Carolyn S. Friedman, Christine Paillard, Jean Laroche & Claude Ferec ### Abstract This protocol permits the mutation scanning of PCR products by high-resolution DNA melting analysis requiring the inclusion of a saturating intercalating dye in the PCR mix without labelled probe. During a scanning process, fluorescent melting curves of PCR amplicons are analyzed. Mutations modifying melting curve shapes, are allowed...

  9. Rapid Detection and Identification of Human Hookworm Infections through High Resolution Melting (HRM) Analysis

    Science.gov (United States)

    Ngui, Romano; Lim, Yvonne A. L.; Chua, Kek Heng

    2012-01-01

    Background Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species. Methods Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples. Conclusion The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species. PMID:22844538

  10. Rapid detection and identification of human hookworm infections through high resolution melting (HRM analysis.

    Directory of Open Access Journals (Sweden)

    Romano Ngui

    Full Text Available BACKGROUND: Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR coupled with high resolution melting-curve (HRM analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species. METHODS: Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2 of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples. CONCLUSION: The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.

  11. High-Resolution Phenotypic Landscape of the RNA Polymerase II Trigger Loop.

    Directory of Open Access Journals (Sweden)

    Chenxi Qiu

    2016-11-01

    Full Text Available The active sites of multisubunit RNA polymerases have a "trigger loop" (TL that multitasks in substrate selection, catalysis, and translocation. To dissect the Saccharomyces cerevisiae RNA polymerase II TL at individual-residue resolution, we quantitatively phenotyped nearly all TL single variants en masse. Three mutant classes, revealed by phenotypes linked to transcription defects or various stresses, have distinct distributions among TL residues. We find that mutations disrupting an intra-TL hydrophobic pocket, proposed to provide a mechanism for substrate-triggered TL folding through destabilization of a catalytically inactive TL state, confer phenotypes consistent with pocket disruption and increased catalysis. Furthermore, allele-specific genetic interactions among TL and TL-proximal domain residues support the contribution of the funnel and bridge helices (BH to TL dynamics. Our structural genetics approach incorporates structural and phenotypic data for high-resolution dissection of transcription mechanisms and their evolution, and is readily applicable to other essential yeast proteins.

  12. Rapid multiplex high resolution melting method to analyze inflammatory related SNPs in preterm birth

    Directory of Open Access Journals (Sweden)

    Pereyra Silvana

    2012-01-01

    Full Text Available Abstract Background Complex traits like cancer, diabetes, obesity or schizophrenia arise from an intricate interaction between genetic and environmental factors. Complex disorders often cluster in families without a clear-cut pattern of inheritance. Genomic wide association studies focus on the detection of tens or hundreds individual markers contributing to complex diseases. In order to test if a subset of single nucleotide polymorphisms (SNPs from candidate genes are associated to a condition of interest in a particular individual or group of people, new techniques are needed. High-resolution melting (HRM analysis is a new method in which polymerase chain reaction (PCR and mutations scanning are carried out simultaneously in a closed tube, making the procedure fast, inexpensive and easy. Preterm birth (PTB is considered a complex disease, where genetic and environmental factors interact to carry out the delivery of a newborn before 37 weeks of gestation. It is accepted that inflammation plays an important role in pregnancy and PTB. Methods Here, we used real time-PCR followed by HRM analysis to simultaneously identify several gene variations involved in inflammatory pathways on preterm labor. SNPs from TLR4, IL6, IL1 beta and IL12RB genes were analyzed in a case-control study. The results were confirmed either by sequencing or by PCR followed by restriction fragment length polymorphism. Results We were able to simultaneously recognize the variations of four genes with similar accuracy than other methods. In order to obtain non-overlapping melting temperatures, the key step in this strategy was primer design. Genotypic frequencies found for each SNP are in concordance with those previously described in similar populations. None of the studied SNPs were associated with PTB. Conclusions Several gene variations related to the same inflammatory pathway were screened through a new flexible, fast and non expensive method with the purpose of analyzing

  13. High-Resolution Melt Analysis for Rapid Comparison of Bacterial Community Compositions

    DEFF Research Database (Denmark)

    Hjelmsø, Mathis Hjort; Hansen, Lars Hestbjerg; Bælum, Jacob

    2014-01-01

    In the study of bacterial community composition, 16S rRNA gene amplicon sequencing is today among the preferred methods of analysis. The cost of nucleotide sequence analysis, including requisite computational and bioinformatic steps, however, takes up a large part of many research budgets. High......-resolution melt (HRM) analysis is the study of the melt behavior of specific PCR products. Here we describe a novel high-throughput approach in which we used HRM analysis targeting the 16S rRNA gene to rapidly screen multiple complex samples for differences in bacterial community composition. We hypothesized...... that HRM analysis of amplified 16S rRNA genes from a soil ecosystem could be used as a screening tool to identify changes in bacterial community structure. This hypothesis was tested using a soil microcosm setup exposed to a total of six treatments representing different combinations of pesticide...

  14. Differentiation of minute virus of mice and mouse parvovirus by high resolution melting curve analysis.

    Science.gov (United States)

    Rao, Dan; Wu, Miaoli; Wang, Jing; Yuan, Wen; Zhu, Yujun; Cong, Feng; Xu, Fengjiao; Lian, Yuexiao; Huang, Bihong; Wu, Qiwen; Chen, Meili; Zhang, Yu; Huang, Ren; Guo, Pengju

    2017-12-01

    Murine parvovirus is one of the most prevalent infectious pathogens in mouse colonies. A specific primer pair targeting the VP2 gene of minute virus of mice (MVM) and mouse parvovirus (MPV) was utilized for high resolution melting (HRM) analysis. The resulting melting curves could distinguish these two virus strains and there was no detectable amplification of the other mouse pathogens which included rat parvovirus (KRV), ectromelia virus (ECT), mouse adenovirus (MAD), mouse cytomegalovirus (MCMV), polyoma virus (Poly), Helicobactor hepaticus (H. hepaticus) and Salmonella typhimurium (S. typhimurium). The detection limit of the standard was 10 copies/μL. This study showed that the PCR-HRM assay could be an alternative useful method with high specificity and sensitivity for differentiating murine parvovirus strains MVM and MPV. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. High-Resolution Melting (HRM) of Hypervariable Mitochondrial DNA Regions for Forensic Science.

    Science.gov (United States)

    Dos Santos Rocha, Alípio; de Amorim, Isis Salviano Soares; Simão, Tatiana de Almeida; da Fonseca, Adenilson de Souza; Garrido, Rodrigo Grazinoli; Mencalha, Andre Luiz

    2018-03-01

    Forensic strategies commonly are proceeding by analysis of short tandem repeats (STRs); however, new additional strategies have been proposed for forensic science. Thus, this article standardized the high-resolution melting (HRM) of DNA for forensic analyzes. For HRM, mitochondrial DNA (mtDNA) from eight individuals were extracted from mucosa swabs by DNAzol reagent, samples were amplified by PCR and submitted to HRM analysis to identify differences in hypervariable (HV) regions I and II. To confirm HRM, all PCR products were DNA sequencing. The data suggest that is possible discriminate DNA from different samples by HRM curves. Also, uncommon dual-dissociation was identified in a single PCR product, increasing HRM analyzes by evaluation of melting peaks. Thus, HRM is accurate and useful to screening small differences in HVI and HVII regions from mtDNA and increase the efficiency of laboratory routines based on forensic genetics. © 2017 American Academy of Forensic Sciences.

  16. Use of a New High Resolution Melting Method for Genotyping Pathogenic Leptospira spp.

    Directory of Open Access Journals (Sweden)

    Florence Naze

    Full Text Available Leptospirosis is a worldwide zoonosis that is endemic in tropical areas, such as Reunion Island. The species Leptospira interrogans is the primary agent in human infections, but other pathogenic species, such as L. kirschner and L. borgpetersenii, are also associated with human leptospirosis.In this study, a melting curve analysis of the products that were amplified with the primer pairs lfb1 F/R and G1/G2 facilitated an accurate species classification of Leptospira reference strains. Next, we combined an unsupervised high resolution melting (HRM method with a new statistical approach using primers to amplify a two variable-number tandem-repeat (VNTR for typing at the subspecies level. The HRM analysis, which was performed with ScreenClust Software, enabled the identification of genotypes at the serovar level with high resolution power (Hunter-Gaston index 0.984. This method was also applied to Leptospira DNA from blood samples that were obtained from Reunion Island after 1998. We were able to identify a unique genotype that is identical to that of the L. interrogans serovars Copenhageni and Icterohaemorrhagiae, suggesting that this genotype is the major cause of leptospirosis on Reunion Island.Our simple, rapid, and robust genotyping method enables the identification of Leptospira strains at the species and subspecies levels and supports the direct genotyping of Leptospira in biological samples without requiring cultures.

  17. Optimization of PCR Condition: The First Study of High Resolution Melting Technique for Screening of APOA1 Variance.

    Science.gov (United States)

    Wahyuningsih, Hesty; K Cayami, Ferdy; Bahrudin, Udin; A Sobirin, Mochamad; Ep Mundhofir, Farmaditya; Mh Faradz, Sultana; Hisatome, Ichiro

    2017-03-01

    High resolution melting (HRM) is a post-PCR technique for variant screening and genotyping based on the different melting points of DNA fragments. The advantages of this technique are that it is fast, simple, and efficient and has a high output, particularly for screening of a large number of samples. APOA1 encodes apolipoprotein A1 (apoA1) which is a major component of high density lipoprotein cholesterol (HDL-C). This study aimed to obtain an optimal quantitative polymerase chain reaction (qPCR)-HRM condition for screening of APOA1 variance. Genomic DNA was isolated from a peripheral blood sample using the salting out method. APOA1 was amplified using the RotorGeneQ 5Plex HRM. The PCR product was visualized with the HRM amplification curve and confirmed using gel electrophoresis. The melting profile was confirmed by looking at the melting curve. Five sets of primers covering the translated region of APOA1 exons were designed with expected PCR product size of 100-400 bps. The amplified segments of DNA were amplicons 2, 3, 4A, 4B, and 4C. Amplicons 2, 3 and 4B were optimized at an annealing temperature of 60 °C at 40 PCR cycles. Amplicon 4A was optimized at an annealing temperature of 62 °C at 45 PCR cycles. Amplicon 4C was optimized at an annealing temperature of 63 °C at 50 PCR cycles. In addition to the suitable procedures of DNA isolation and quantification, primer design and an estimated PCR product size, the data of this study showed that appropriate annealing temperature and PCR cycles were important factors in optimization of HRM technique for variant screening in APOA1 .

  18. High-resolution melting-curve (HRM) analysis for C. meleagridis identification in stool samples.

    Science.gov (United States)

    Chelbi, Hanen; Essid, Rym; Jelassi, Refka; Bouzekri, Nesrine; Zidi, Ines; Ben Salah, Hamza; Mrad, Ilhem; Ben Sghaier, Ines; Abdelmalek, Rym; Aissa, Sameh; Bouratbine, Aida; Aoun, Karim

    2018-02-01

    Cryptosporidiosis represents a major public health problem. This infection, caused by a protozoan parasite of the genus Cryptosporidium, has been reported worldwide as a frequent cause of diarrhoea. In the immunocompetent host, the typical watery diarrhea can be self-limiting. However, it is severe and chronic, in the immunocompromised host and may cause death. Cryptosporidium spp. are coccidians, which complete their life cycle in both humans and animals. The two species C. hominis and C. parvum are the major cause of human infection. Compared to studies on C. hominis and C. parvum, only a few studies have developed methods to identify C. meleagridis. To develop a new real time PCR-coupled High resolution melting assay allowing the detection for C. meleagridis, in addition of the other dominant species (C. hominis and C. parvum). The polymorphic sequence on the dihydrofolate reductase gene (DHFR) of three species was sequenced to design primers pair and establish a sensitive real-time PCR coupled to a high-resolution melting-curve (HRM) analysis method, allowing the detection of Cryptosporidium sp. and discrimination between three prevalent species in Tunisia. We analyzed a collection of 42 archived human isolates of the three studied species. Real-time PCR coupled to HRM assay allowed detection of Cryptosporidium, using the new designed primers, and basing on melting profile, we can distinguish C. meleagridis species in addition to C. parvum and C. hominis. We developed a qPCR-HRM assay that allows Cryptosporidium genotyping. This method is sensitive and able to distinguish three Cryptosporidium species. Copyright © 2017. Published by Elsevier Ltd.

  19. High resolution melt curve analysis based on methylation status for human semen identification.

    Science.gov (United States)

    Fachet, Caitlyn; Quarino, Lawrence; Karnas, K Joy

    2017-03-01

    A high resolution melt curve assay to differentiate semen from blood, saliva, urine, and vaginal fluid based on methylation status at the Dapper Isoform 1 (DACT1) gene was developed. Stains made from blood, saliva, urine, semen, and vaginal fluid were obtained from volunteers and DNA was isolated using either organic extraction (saliva, urine, and vaginal fluid) or Chelex ® 100 extraction (blood and semen). Extracts were then subjected to bisulfite modification in order to convert unmethylated cytosines to uracil, consequently creating sequences whose amplicons have melt curves that vary depending on their initial methylation status. When primers designed to amplify the promoter region of the DACT1 gene were used, DNA from semen samples was distinguishable from other fluids by a having a statistically significant lower melting temperature. The assay was found to be sperm-significant since semen from a vasectomized man produced a melting temperature similar to the non-semen body fluids. Blood and semen stains stored up to 5 months and tested at various intervals showed little variation in melt temperature indicating the methylation status was stable during the course of the study. The assay is a more viable method for forensic science practice than most molecular-based methods for body fluid stain identification since it is time efficient and utilizes instrumentation common to forensic biology laboratories. In addition, the assay is advantageous over traditional presumptive chemical methods for body fluid identification since results are confirmatory and the assay offers the possibility of multiplexing which may test for multiple body fluids simultaneously.

  20. The spa typing of methicillin-resistant Staphylococcus aureus isolates by High Resolution Melting (HRM) analysis.

    Science.gov (United States)

    Fasihi, Yasser; Fooladi, Saba; Mohammadi, Mohammad Ali; Emaneini, Mohammad; Kalantar-Neyestanaki, Davood

    2017-09-06

    Molecular typing is an important tool for control and prevention of infection. A suitable molecular typing method for epidemiological investigation must be easy to perform, highly reproducible, inexpensive, rapid and easy to interpret. In this study, two molecular typing methods including the conventional PCR-sequencing method and high resolution melting (HRM) analysis were used for staphylococcal protein A (spa) typing of 30 Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from clinical samples. Based on PCR-sequencing method results, 16 different spa types were identified among the 30 MRSA isolates. Among the 16 different spa types, 14 spa types separated by HRM method. Two spa types including t4718 and t2894 were not separated from each other. According to our results, spa typing based on HRM analysis method is very rapid, easy to perform and cost-effective, but this method must be standardized for different regions, spa types, and real-time machinery.

  1. High-resolution melt analysis for species identification of coagulase-negative staphylococci derived from bovine milk.

    Science.gov (United States)

    Ajitkumar, Praseeda; Barkema, Herman W; Zadoks, Ruth N; Morck, Douglas W; van der Meer, Frank J U M; De Buck, Jeroen

    2013-03-01

    Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens isolated from bovine milk. In this study, we report a rapid assay for species identification of CNS using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time polymerase chain reaction amplification of 16S rRNA gene fragment, spanning the variable region V1 and V2, was performed with a resulting amplicon of 215 bp. A library of distinct melt curves of reference strains of 13 common CNS species was created using HRMA. Sequencing of 16S rRNA and rpoB genes, and, when needed, tuf gene, of 100 CNS isolates obtained from Canadian Bovine Mastitis Research Network was done to determine their species identity, allowing for subsequent evaluation of the performance of HRMA for field isolates of bovine CNS. A combination of HRMA and sequencing revealed that Staphylococcus chromogenes, S. xylosus, S. simulans, and S. sciuri had multiple genotypes, complicating their resolution by HRMA. As the 3 genotypes of S. chromogenes had distinct melt curves, the 3 distinct genotypes were employed as reference strains in a blinded trial of 156 CNS isolates to identify S. chromogenes. HRMA correctly identified all S. chromogenes isolates which were later confirmed by sequencing. Staphylococcus chromogenes (68%) was most frequently found among the CNS isolates, followed by S. haemolyticus (10%) and S. xylosus (6%). The present study revealed that HRMA of 16S rRNA gene (V1-V2) could be used as a rapid, efficient, low-cost, and minimally cumbersome technique for S. chromogenes identification, the most common CNS derived from bovine milk. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Genome-wide SNP discovery in tetraploid alfalfa using 454 sequencing and high resolution melting analysis

    Directory of Open Access Journals (Sweden)

    Zhao Patrick X

    2011-07-01

    Full Text Available Abstract Background Single nucleotide polymorphisms (SNPs are the most common type of sequence variation among plants and are often functionally important. We describe the use of 454 technology and high resolution melting analysis (HRM for high throughput SNP discovery in tetraploid alfalfa (Medicago sativa L., a species with high economic value but limited genomic resources. Results The alfalfa genotypes selected from M. sativa subsp. sativa var. 'Chilean' and M. sativa subsp. falcata var. 'Wisfal', which differ in water stress sensitivity, were used to prepare cDNA from tissue of clonally-propagated plants grown under either well-watered or water-stressed conditions, and then pooled for 454 sequencing. Based on 125.2 Mb of raw sequence, a total of 54,216 unique sequences were obtained including 24,144 tentative consensus (TCs sequences and 30,072 singletons, ranging from 100 bp to 6,662 bp in length, with an average length of 541 bp. We identified 40,661 candidate SNPs distributed throughout the genome. A sample of candidate SNPs were evaluated and validated using high resolution melting (HRM analysis. A total of 3,491 TCs harboring 20,270 candidate SNPs were located on the M. truncatula (MT 3.5.1 chromosomes. Gene Ontology assignments indicate that sequences obtained cover a broad range of GO categories. Conclusions We describe an efficient method to identify thousands of SNPs distributed throughout the alfalfa genome covering a broad range of GO categories. Validated SNPs represent valuable molecular marker resources that can be used to enhance marker density in linkage maps, identify potential factors involved in heterosis and genetic variation, and as tools for association mapping and genomic selection in alfalfa.

  3. High resolution melting for mutation scanning of TP53 exons 5–8

    International Nuclear Information System (INIS)

    Krypuy, Michael; Dobrovic, Alexander; Ahmed, Ahmed Ashour; Etemadmoghadam, Dariush; Hyland, Sarah J; Australian Ovarian Cancer Study Group; Fazio, Anna de; Fox, Stephen B; Brenton, James D; Bowtell, David D

    2007-01-01

    p53 is commonly inactivated by mutations in the DNA-binding domain in a wide range of cancers. As mutant p53 often influences response to therapy, effective and rapid methods to scan for mutations in TP53 are likely to be of clinical value. We therefore evaluated the use of high resolution melting (HRM) as a rapid mutation scanning tool for TP53 in tumour samples. We designed PCR amplicons for HRM mutation scanning of TP53 exons 5 to 8 and tested them with DNA from cell lines hemizygous or homozygous for known mutations. We assessed the sensitivity of each PCR amplicon using dilutions of cell line DNA in normal wild-type DNA. We then performed a blinded assessment on ovarian tumour DNA samples that had been previously sequenced for mutations in TP53 to assess the sensitivity and positive predictive value of the HRM technique. We also performed HRM analysis on breast tumour DNA samples with unknown TP53 mutation status. One cell line mutation was not readily observed when exon 5 was amplified. As exon 5 contained multiple melting domains, we divided the exon into two amplicons for further screening. Sequence changes were also introduced into some of the primers to improve the melting characteristics of the amplicon. Aberrant HRM curves indicative of TP53 mutations were observed for each of the samples in the ovarian tumour DNA panel. Comparison of the HRM results with the sequencing results revealed that each mutation was detected by HRM in the correct exon. For the breast tumour panel, we detected seven aberrant melt profiles by HRM and subsequent sequencing confirmed the presence of these and no other mutations in the predicted exons. HRM is an effective technique for simple and rapid scanning of TP53 mutations that can markedly reduce the amount of sequencing required in mutational studies of TP53

  4. High resolution melting analysis (HRM) for the assessment of clonality in feline B-cell lymphomas.

    Science.gov (United States)

    Henrich, Manfred; Scheffold, Svenja; Hecht, Werner; Reinacher, Manfred

    2018-06-01

    Analysis of clonality is gaining importance in diagnosing lymphomas in veterinary medicine. Usually, PCR for the analysis of antigen receptor rearrangement (PARR) is followed by electrophoretic separation of the PCR products. Aim of this study was to test the feasibility of HRM for the assessment of clonality in B-cell lymphomas of cats. High resolution melting analysis differentiates PCR products by their different melting point using the decrease in fluorescence of an intercalating dye during melting of the PCR product. Additionally, the method is easy to use with no post-PCR manipulation of the samples. Forty-seven feline B-cell lymphomas and 31 reactive lymphatic proliferations of cats were investigated by PARR followed either by capillary electrophoresis or an HRM assay. To objectify the interpretation of the HRM results a recently published mathematical approach was applied to the melting curve. To overcome discrepancies between the visual interpretation and the mathematical approach, the latter was modified to include testing of reproducibility and recognition of pseudoclonality. In 11 of 47 lymphoma cases clonal populations were detectable by HRM assay compared to 14 of 47 lymphomas in which clonal populations were detected by capillary electrophoresis assay. Neither of the methods showed a clonal pattern in any of the reactive samples. However, the HRM assay showed a unique pattern in cases of follicular lymphatic hyperplasia that had no corresponding pattern in capillary electrophoresis. The capillary electrophoresis assay could identify 3 lymphomas that were not detected by the HRM assay and is therefore regarded superior to the HRM assay. The comparison however, was hampered by the overall bad performance of the PARR, that might be the consequence of insufficient primer binding due to somatic hypermutation of the binding sites during antigen stimulated proliferation of the B lymphocytes. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Evaluation of PCR and high-resolution melt curve analysis for differentiation of Salmonella isolates.

    Science.gov (United States)

    Saeidabadi, Mohammad Sadegh; Nili, Hassan; Dadras, Habibollah; Sharifiyazdi, Hassan; Connolly, Joanne; Valcanis, Mary; Raidal, Shane; Ghorashi, Seyed Ali

    2017-06-01

    Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the serovar/serotype.

  6. Applying high-resolution melting (HRM) technology to identify five commonly used Artemisia species.

    Science.gov (United States)

    Song, Ming; Li, Jingjian; Xiong, Chao; Liu, Hexia; Liang, Junsong

    2016-10-04

    Many members of the genus Artemisia are important for medicinal purposes with multiple pharmacological properties. Often, these herbal plants sold on the markets are in processed forms so it is difficult to authenticate. Routine testing and identification of these herbal materials should be performed to ensure that the raw materials used in pharmaceutical products are suitable for their intended use. In this study, five commonly used Artemisia species included Artemisia argyi, Artemisia annua, Artemisia lavandulaefolia, Artemisia indica, and Artemisia atrovirens were analyzed using high resolution melting (HRM) analysis based on the internal transcribed spacer 2 (ITS2) sequences. The melting profiles of the ITS2 amplicons of the five closely related herbal species are clearly separated so that they can be differentiated by HRM method. The method was further applied to authenticate commercial products in powdered. HRM curves of all the commercial samples tested are similar to the botanical species as labeled. These congeneric medicinal products were also clearly separated using the neighbor-joining (NJ) tree. Therefore, HRM method could provide an efficient and reliable authentication system to distinguish these commonly used Artemisia herbal products on the markets and offer a technical reference for medicines quality control in the drug supply chain.

  7. Rapid detection and identification of four major Schistosoma species by high-resolution melt (HRM) analysis.

    Science.gov (United States)

    Li, Juan; Zhao, Guang-Hui; Lin, RuiQing; Blair, David; Sugiyama, Hiromu; Zhu, Xing-Quan

    2015-11-01

    Schistosomiasis, caused by blood flukes belonging to several species of the genus Schistosoma, is a serious and widespread parasitic disease. Accurate and rapid differentiation of these etiological agents of animal and human schistosomiasis to species level can be difficult. We report a real-time PCR assay coupled with a high-resolution melt (HRM) assay targeting a portion of the nuclear 18S rDNA to detect, identify, and distinguish between four major blood fluke species (Schistosoma japonicum, Schistosoma mansoni, Schistosoma haematobium, and Schistosoma mekongi). Using this system, the Schistosoma spp. was accurately identified and could also be distinguished from all other trematode species with which they were compared. As little as 10(-5) ng genomic DNA from a Schistosoma sp. could be detected. This process is inexpensive, easy, and can be completed within 3 h. Examination of 21 representative Schistosoma samples from 15 geographical localities in seven endemic countries validated the value of the HRM detection assay and proved its reliability. The melting curves were characterized by peaks of 83.65 °C for S. japonicum and S. mekongi, 85.65 °C for S. mansoni, and 85.85 °C for S. haematobium. The present study developed a real-time PCR coupled with HRM analysis assay for detection and differential identification of S. mansoni, S. haematobium, S. japonicum, and S. mekongi. This method is rapid, sensitive, and inexpensive. It has important implications for epidemiological studies of Schistosoma.

  8. Massively parallel digital high resolution melt for rapid and absolutely quantitative sequence profiling

    Science.gov (United States)

    Velez, Daniel Ortiz; Mack, Hannah; Jupe, Julietta; Hawker, Sinead; Kulkarni, Ninad; Hedayatnia, Behnam; Zhang, Yang; Lawrence, Shelley; Fraley, Stephanie I.

    2017-02-01

    In clinical diagnostics and pathogen detection, profiling of complex samples for low-level genotypes represents a significant challenge. Advances in speed, sensitivity, and extent of multiplexing of molecular pathogen detection assays are needed to improve patient care. We report the development of an integrated platform enabling the identification of bacterial pathogen DNA sequences in complex samples in less than four hours. The system incorporates a microfluidic chip and instrumentation to accomplish universal PCR amplification, High Resolution Melting (HRM), and machine learning within 20,000 picoliter scale reactions, simultaneously. Clinically relevant concentrations of bacterial DNA molecules are separated by digitization across 20,000 reactions and amplified with universal primers targeting the bacterial 16S gene. Amplification is followed by HRM sequence fingerprinting in all reactions, simultaneously. The resulting bacteria-specific melt curves are identified by Support Vector Machine learning, and individual pathogen loads are quantified. The platform reduces reaction volumes by 99.995% and achieves a greater than 200-fold increase in dynamic range of detection compared to traditional PCR HRM approaches. Type I and II error rates are reduced by 99% and 100% respectively, compared to intercalating dye-based digital PCR (dPCR) methods. This technology could impact a number of quantitative profiling applications, especially infectious disease diagnostics.

  9. Reliable discrimination of 10 ungulate species using high resolution melting analysis of faecal DNA.

    Directory of Open Access Journals (Sweden)

    Ana Ramón-Laca

    Full Text Available Identifying species occupying an area is essential for many ecological and conservation studies. Faecal DNA is a potentially powerful method for identifying cryptic mammalian species. In New Zealand, 10 species of ungulate (Order: Artiodactyla have established wild populations and are managed as pests because of their impacts on native ecosystems. However, identifying the ungulate species present within a management area based on pellet morphology is unreliable. We present a method that enables reliable identification of 10 ungulate species (red deer, sika deer, rusa deer, fallow deer, sambar deer, white-tailed deer, Himalayan tahr, Alpine chamois, feral sheep, and feral goat from swabs of faecal pellets. A high resolution melting (HRM assay, targeting a fragment of the 12S rRNA gene, was developed. Species-specific primers were designed and combined in a multiplex PCR resulting in fragments of different length and therefore different melting behaviour for each species. The method was developed using tissue from each of the 10 species, and was validated in blind trials. Our protocol enabled species to be determined for 94% of faecal pellet swabs collected during routine monitoring by the New Zealand Department of Conservation. Our HRM method enables high-throughput and cost-effective species identification from low DNA template samples, and could readily be adapted to discriminate other mammalian species from faecal DNA.

  10. Rapid detection of Echinococcus species by a high-resolution melting (HRM) approach.

    Science.gov (United States)

    Santos, Guilherme Brzoskowski; Espínola, Sergio Martín; Ferreira, Henrique Bunselmeyer; Margis, Rogerio; Zaha, Arnaldo

    2013-11-14

    High-resolution melting (HRM) provides a low-cost, fast and sensitive scanning method that allows the detection of DNA sequence variations in a single step, which makes it appropriate for application in parasite identification and genotyping. The aim of this work was to implement an HRM-PCR assay targeting part of the mitochondrial cox1 gene to achieve an accurate and fast method for Echinococcus spp. differentiation. For melting analysis, a total of 107 samples from seven species were used in this study. The species analyzed included Echinococcus granulosus (n = 41) and Echinococcus ortleppi (n = 50) from bovine, Echinococcus vogeli (n = 2) from paca, Echinococcus oligarthra (n = 3) from agouti, Echinococcus multilocularis (n = 6) from monkey and Echinococcus canadensis (n = 2) and Taenia hydatigena (n = 3) from pig. DNA extraction was performed, and a 444-bp fragment of the cox1 gene was amplified. Two approaches were used, one based on HRM analysis, and a second using SYBR Green Tm-based. In the HRM analysis, a specific profile for each species was observed. Although some species exhibited almost the same melting temperature (Tm) value, the HRM profiles could be clearly discriminated. The SYBR Green Tm-based analysis showed differences between E. granulosus and E. ortleppi and between E. vogeli and E. oligarthra. In this work, we report the implementation of HRM analysis to differentiate species of the genus Echinococcus using part of the mitochondrial gene cox1. This method may be also potentially applied to identify other species belonging to the Taeniidae family.

  11. Differentiation of five enterohepatic Helicobacter species by nested PCR with high-resolution melting curve analysis.

    Science.gov (United States)

    Wu, Miaoli; Rao, Dan; Zhu, Yujun; Wang, Jing; Yuan, Wen; Zhang, Yu; Huang, Ren; Guo, Pengju

    2017-04-01

    Enterohepatic Helicobacter species (EHS) are widespread in rodent species around the world. Several studies have demonstrated that infection with EHS can interfere with the outcomes of animal experiments in cancer research and significantly influence the study results. Therefore, it is essential to establish a rapid detection and identification of EHS for biomedical research using laboratory rodents. Our study aimed to develop a rapid and sensitive method to detect and distinguish five enterohepatic Helicobacter species. Nested PCR followed by high-resolution melting curve analysis (HRM) was developed for identification of H. bilis, H. rodentium, H. muridarum, H. typhlonius, as well as H. hepaticus. To validate the accuracy of nested PCR-HRM analysis, quantitative real-time PCR methods for five different enterohepatic Helicobacter species were developed. A total of 50 cecal samples were tested using both nested PCR-HRM analysis and qPCR method. The nested PCR-HRM method could distinguish five enterohepatic Helicobacter species by different melting temperatures. The melting curve were characterized by peaks of 78.7 ± 0.12°C for H. rodentium, 80.51 ± 0.09°C for H. bilis, 81.6 ± 0.1°C for H. typhlonius, 82.11 ± 0.18°C for H. muridarum, and 82.95 ± 0.09°C for H. hepaticus. The nested PCR-HRM assay is a simple, rapid, and cost-effective assay. This assay could be a useful tool for molecular epidemiology study of enterohepatic Helicobacter infection and an attractive alternative for genotyping of enterohepatic Helicobacter species. © 2016 John Wiley & Sons Ltd.

  12. High-resolution melt PCR analysis for genotyping of Ureaplasma parvum isolates directly from clinical samples.

    Science.gov (United States)

    Payne, Matthew S; Tabone, Tania; Kemp, Matthew W; Keelan, Jeffrey A; Spiller, O Brad; Newnham, John P

    2014-02-01

    Ureaplasma sp. infection in neonates and adults underlies a variety of disease pathologies. Of the two human Ureaplasma spp., Ureaplasma parvum is clinically the most common. We have developed a high-resolution melt (HRM) PCR assay for the differentiation of the four serovars of U. parvum in a single step. Currently U. parvum strains are separated into four serovars by sequencing the promoter and coding region of the multiple-banded antigen (MBA) gene. We designed primers to conserved sequences within this region for PCR amplification and HRM analysis to generate reproducible and distinct melt profiles that distinguish clonal representatives of serovars 1, 3, 6, and 14. Furthermore, our HRM PCR assay could classify DNA extracted from 74 known (MBA-sequenced) test strains with 100% accuracy. Importantly, HRM PCR was also able to identify U. parvum serovars directly from 16 clinical swabs. HRM PCR performed with DNA consisting of mixtures of combined known serovars yielded profiles that were easily distinguished from those for single-serovar controls. These profiles mirrored clinical samples that contained mixed serovars. Unfortunately, melt curve analysis software is not yet robust enough to identify the composition of mixed serovar samples, only that more than one serovar is present. HRM PCR provides a single-step, rapid, cost-effective means to differentiate the four serovars of U. parvum that did not amplify any of the known 10 serovars of Ureaplasma urealyticum tested in parallel. Choice of reaction reagents was found to be crucial to allow sufficient sensitivity to differentiate U. parvum serovars directly from clinical swabs rather than requiring cell enrichment using microbial culture techniques.

  13. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.

    Science.gov (United States)

    Zampieri, Ricardo Andrade; Laranjeira-Silva, Maria Fernanda; Muxel, Sandra Marcia; Stocco de Lima, Ana Carolina; Shaw, Jeffrey Jon; Floeter-Winter, Lucile Maria

    2016-02-01

    Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.

  14. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.

    Directory of Open Access Journals (Sweden)

    Ricardo Andrade Zampieri

    2016-02-01

    Full Text Available Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol.Exploring the High Resolution Melting (HRM dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR targeting heat-shock protein 70 coding gene (hsp70 revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania infantum chagasi, L. (L. amazonensis, L. (L. mexicana, L. (Viannia lainsoni, L. (V. braziliensis, L. (V. guyanensis, L. (V. naiffi and L. (V. shawi, and three species found in Eurasia and Africa, including L. (L. tropica, L. (L. donovani and L. (L. major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol.HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.

  15. Rapid screening for targeted genetic variants via high-resolution melting curve analysis.

    Science.gov (United States)

    Chambliss, Allison B; Resnick, Molly; Petrides, Athena K; Clarke, William A; Marzinke, Mark A

    2017-03-01

    Current methods for the detection of single nucleotide polymorphisms (SNPs) associated with aberrant drug-metabolizing enzyme function are hindered by long turnaround times and specialized techniques and instrumentation. In this study, we describe the development and validation of a high-resolution melting (HRM) curve assay for the rapid screening of variant genotypes for targeted genetic polymorphisms in the cytochrome P450 enzymes CYP2C9, CYP2C19, and CYP3A5. Sequence-specific primers were custom-designed to flank nine SNPs within the genetic regions of aforementioned drug metabolizing enzymes. PCR amplification was performed followed by amplicon denaturation by precise temperature ramping in order to distinguish genotypes by melting temperature (Tm). A standardized software algorithm was used to assign amplicons as 'reference' or 'variant' as compared to duplicate reference sequence DNA controls for each SNP. Intra-assay (n=5) precision of Tms for all SNPs was ≤0.19%, while inter-assay (n=20) precision ranged from 0.04% to 0.21%. When compared to a reference method of Sanger sequencing, the HRM assay produced no false negative results, and overcall frequency ranged from 0% to 26%, depending on the SNP. Furthermore, HRM genotyping displayed accuracy over input DNA concentrations ranging from 10 to 200 ng/μL. The presented assay provides a rapid method for the screening for genetic variants in targeted CYP450 regions with a result of 'reference' or 'variant' available within 2 h from receipt of extracted DNA. The method can serve as a screening approach to rapidly identify individuals with variant sequences who should be further investigated by reflexed confirmatory testing for aberrant cytochrome P450 enzymatic activity. Rapid knowledge of variant status may aid in the avoidance of adverse clinical events by allowing for dosing of normal metabolizer patients immediately while identifying the need to wait for confirmatory testing in those patients who are

  16. Rapid detection of SMARCB1 sequence variation using high resolution melting

    Directory of Open Access Journals (Sweden)

    Ashley David M

    2009-12-01

    Full Text Available Abstract Background Rhabdoid tumors are rare cancers of early childhood arising in the kidney, central nervous system and other organs. The majority are caused by somatic inactivating mutations or deletions affecting the tumor suppressor locus SMARCB1 [OMIM 601607]. Germ-line SMARCB1 inactivation has been reported in association with rhabdoid tumor, epitheloid sarcoma and familial schwannomatosis, underscoring the importance of accurate mutation screening to ascertain recurrence and transmission risks. We describe a rapid and sensitive diagnostic screening method, using high resolution melting (HRM, for detecting sequence variations in SMARCB1. Methods Amplicons, encompassing the nine coding exons of SMARCB1, flanking splice site sequences and the 5' and 3' UTR, were screened by both HRM and direct DNA sequencing to establish the reliability of HRM as a primary mutation screening tool. Reaction conditions were optimized with commercially available HRM mixes. Results The false negative rate for detecting sequence variants by HRM in our sample series was zero. Nine amplicons out of a total of 140 (6.4% showed variant melt profiles that were subsequently shown to be false positive. Overall nine distinct pathogenic SMARCB1 mutations were identified in a total of 19 possible rhabdoid tumors. Two tumors had two distinct mutations and two harbored SMARCB1 deletion. Other mutations were nonsense or frame-shifts. The detection sensitivity of the HRM screening method was influenced by both sequence context and specific nucleotide change and varied from 1: 4 to 1:1000 (variant to wild-type DNA. A novel method involving digital HRM, followed by re-sequencing, was used to confirm mutations in tumor specimens containing associated normal tissue. Conclusions This is the first report describing SMARCB1 mutation screening using HRM. HRM is a rapid, sensitive and inexpensive screening technology that is likely to be widely adopted in diagnostic laboratories to

  17. Rapid detection of SMARCB1 sequence variation using high resolution melting

    International Nuclear Information System (INIS)

    Dagar, Vinod; Chow, Chung-Wo; Ashley, David M; Algar, Elizabeth M

    2009-01-01

    Rhabdoid tumors are rare cancers of early childhood arising in the kidney, central nervous system and other organs. The majority are caused by somatic inactivating mutations or deletions affecting the tumor suppressor locus SMARCB1 [OMIM 601607]. Germ-line SMARCB1 inactivation has been reported in association with rhabdoid tumor, epitheloid sarcoma and familial schwannomatosis, underscoring the importance of accurate mutation screening to ascertain recurrence and transmission risks. We describe a rapid and sensitive diagnostic screening method, using high resolution melting (HRM), for detecting sequence variations in SMARCB1. Amplicons, encompassing the nine coding exons of SMARCB1, flanking splice site sequences and the 5' and 3' UTR, were screened by both HRM and direct DNA sequencing to establish the reliability of HRM as a primary mutation screening tool. Reaction conditions were optimized with commercially available HRM mixes. The false negative rate for detecting sequence variants by HRM in our sample series was zero. Nine amplicons out of a total of 140 (6.4%) showed variant melt profiles that were subsequently shown to be false positive. Overall nine distinct pathogenic SMARCB1 mutations were identified in a total of 19 possible rhabdoid tumors. Two tumors had two distinct mutations and two harbored SMARCB1 deletion. Other mutations were nonsense or frame-shifts. The detection sensitivity of the HRM screening method was influenced by both sequence context and specific nucleotide change and varied from 1: 4 to 1:1000 (variant to wild-type DNA). A novel method involving digital HRM, followed by re-sequencing, was used to confirm mutations in tumor specimens containing associated normal tissue. This is the first report describing SMARCB1 mutation screening using HRM. HRM is a rapid, sensitive and inexpensive screening technology that is likely to be widely adopted in diagnostic laboratories to facilitate whole gene mutation screening

  18. Diagnosis of genetic predisposition for lactose intolerance by high resolution melting analysis.

    Science.gov (United States)

    Delacour, Hervé; Leduc, Amandine; Louçano-Perdriat, Andréa; Plantamura, Julie; Ceppa, Franck

    2017-02-01

    Lactose, the principle sugar in milk, is a disaccharide hydrolyzed by intestinal lactase into glucose and galactose, which are absorbed directly by diffusion in the intestine. The decline of lactase expression (or hypolactasia) in intestinal microvilli after weaning is a normal phenomenon in mammals known as lactase deficiency. It is observed in nearly 75% of the world population and is an inherited autosomal recessive trait with incomplete penetrance. It is caused by SNPs in a regulatory element for lactase gene. In Indo-European, lactase deficiency is associated with rs4982235 SNP (or -13910C>T). The aim of this study is to describe a method based on high resolution melting for rapidly detecting genetic predisposition to lactose intolerance. Analytical performance of the assay was assessed by evaluating within and betwwen-run precision and by comparing the results (n = 50 patients) obtained with the HRM assay to those obtained with the gold standard (Sanger sequencing of the region of interest). In silico prediction of HRM curves was performed to evaluate the potential impact of the other SNPs described within the PCR product on the HRM analytical performances. The assay has good performance (CV lactose intolerance.

  19. Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species.

    Directory of Open Access Journals (Sweden)

    Maslin Osathanunkul

    Full Text Available DNA barcoding coupled high resolution melting (Bar-HRM is an emerging method for species discrimination based on DNA dissociation kinetics. The aim of this work was to evaluate the suitability of different primer sets, derived from selected DNA regions, for Bar-HRM analysis of species in Croton (Euphorbiaceae, one of the largest genera of plants with over 1,200 species. Seven primer pairs were evaluated (matK, rbcL1, rbcL2, rbcL3, rpoC, trnL and ITS1 from four plastid regions, matK, rbcL, rpoC, and trnL, and the nuclear ribosomal marker ITS1. The primer pair derived from the ITS1 region was the single most effective region for the identification of the tested species, whereas the rbcL1 primer pair gave the lowest resolution. It was observed that the ITS1 barcode was the most useful DNA barcoding region overall for species discrimination out of all of the regions and primers assessed. Our Bar-HRM results here also provide further support for the hypothesis that both sequence and base composition affect DNA duplex stability.

  20. Detection of somatic mutations by high-resolution DNA melting (HRM) analysis in multiple cancers.

    Science.gov (United States)

    Gonzalez-Bosquet, Jesus; Calcei, Jacob; Wei, Jun S; Garcia-Closas, Montserrat; Sherman, Mark E; Hewitt, Stephen; Vockley, Joseph; Lissowska, Jolanta; Yang, Hannah P; Khan, Javed; Chanock, Stephen

    2011-01-17

    Identification of somatic mutations in cancer is a major goal for understanding and monitoring the events related to cancer initiation and progression. High resolution melting (HRM) curve analysis represents a fast, post-PCR high-throughput method for scanning somatic sequence alterations in target genes. The aim of this study was to assess the sensitivity and specificity of HRM analysis for tumor mutation screening in a range of tumor samples, which included 216 frozen pediatric small rounded blue-cell tumors as well as 180 paraffin-embedded tumors from breast, endometrial and ovarian cancers (60 of each). HRM analysis was performed in exons of the following candidate genes known to harbor established commonly observed mutations: PIK3CA, ERBB2, KRAS, TP53, EGFR, BRAF, GATA3, and FGFR3. Bi-directional sequencing analysis was used to determine the accuracy of the HRM analysis. For the 39 mutations observed in frozen samples, the sensitivity and specificity of HRM analysis were 97% and 87%, respectively. There were 67 mutation/variants in the paraffin-embedded samples, and the sensitivity and specificity for the HRM analysis were 88% and 80%, respectively. Paraffin-embedded samples require higher quantity of purified DNA for high performance. In summary, HRM analysis is a promising moderate-throughput screening test for mutations among known candidate genomic regions. Although the overall accuracy appears to be better in frozen specimens, somatic alterations were detected in DNA extracted from paraffin-embedded samples.

  1. The high resolution melting analysis (HRM) as a molecular tool for monitoring parasites of the wildlife.

    Science.gov (United States)

    Héritier, Laurent; Verneau, Olivier; Breuil, Gregory; Meistertzheim, Anne-Leila

    2017-04-01

    In an interconnected world, the international pet trade on wild animals is becoming increasingly important. As a consequence, non-native parasite species are introduced, which affect the health of wildlife and contribute to the loss of biodiversity. Because the investigation of parasite diversity within vulnerable host species implies the molecular identification of large samples of parasite eggs, the sequencing of DNA barcodes is time-consuming and costly. Thereby, the objectives of our study were to apply the high resolution melting (HRM) approach for species determination from pools of parasite eggs. Molecular assays were validated on flatworm parasites (polystomes) infecting the Mediterranean pond turtle Mauremys leprosa and the invasive red-eared slider Trachemys scripta elegans in French natural environments. HRM analysis results indicated that double or multiple parasitic infections could be detected from wild animal populations. They also showed that the cycle of parasite eggs production was not regular over time and may depend on several factors, among which the ecological niche and the target species. Thereby, monitoring parasites from wild endangered animals implies periodic parasitological surveys to avoid false negative diagnostics, based solely on eggs production.

  2. Applying high-resolution melting (HRM) technology to olive oil and wine authenticity.

    Science.gov (United States)

    Pereira, Leonor; Gomes, Sónia; Barrias, Sara; Fernandes, José Ramiro; Martins-Lopes, Paula

    2018-01-01

    Olive oil and wine production have a worldwide economic impact. Their market reliability is under great concern because of the increasing number of fraud and adulteration attempts. The need for a traceability system in all its extension is crucial particularly for the cases of olive oils and wines with certified labels, in which only a limited number of olives and grapevine varieties, respectively, are allowed in a restricted well-defined geographical area. Molecular markers have been vastly applied to the food sector, and in particular High-Resolution DNA Melting technology has been successfully applied for olive oil and wine authentication, as part of the traceability system. In this review, the applications of HRM and their usefulness for this sector considering, Safety, Security and Authenticity will be reviewed. A broad overview of the HRM technique will be presented, focusing on the aspects that are crucial for its success, in particular the new generation of fluorescent dsDNA dyes used for amplicon detection and quantification, and the data analysis. A brief outlook on the olive oil and wine authenticity procedures, based on new DNA technology advances, and in which way this may influence the future establishment of a traceability system will be discussed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. High-resolution melting (HRM) for genotyping bovine ephemeral fever virus (BEFV).

    Science.gov (United States)

    Erster, Oran; Stram, Rotem; Menasherow, Shopia; Rubistein-Giuni, Marisol; Sharir, Binyamin; Kchinich, Evgeni; Stram, Yehuda

    2017-02-02

    In recent years there have been several major outbreaks of bovine ephemeral disease in the Middle East, including Israel. Such occurrences raise the need for quick identification of the viruses responsible for the outbreaks, in order to rapidly identify the entry of viruses that do not belong to the Middle-East BEFV lineage. This challenge was met by the development of a high-resolution melt (HRM) assay. The assay is based on the viral G gene sequence and generation of an algorithm that calculates and evaluates the GC content of various fragments. The algorithm was designed to scan 50- to 200-base-long segments in a sliding-window manner, compare and rank them using an Order of Technique of Preference by Similarity to Ideal Solution (TOPSIS) the technique for order preference by similarity to ideal solution technique, according to the differences in GC content of homologous fragments. Two fragments were selected, based on a match to the analysis criteria, in terms of size and GC content. These fragments were successfully used in the analysis to differentiate between different virus lineages, thus facilitating assignment of the viruses' geographical origins. Moreover, the assay could be used for differentiating infected from vaccinated animales (DIVA). The new algorithm may therefore be useful for development of improved genotyping studies for other viruses and possibly other microorganisms. Copyright © 2016. Published by Elsevier B.V.

  4. Identification of Uvaria sp by barcoding coupled with high-resolution melting analysis (Bar-HRM).

    Science.gov (United States)

    Osathanunkul, M; Madesis, P; Ounjai, S; Pumiputavon, K; Somboonchai, R; Lithanatudom, P; Chaowasku, T; Wipasa, J; Suwannapoom, C

    2016-01-13

    DNA barcoding, which was developed about a decade ago, relies on short, standardized regions of the genome to identify plant and animal species. This method can be used to not only identify known species but also to discover novel ones. Numerous sequences are stored in online databases worldwide. One of the ways to save cost and time (by omitting the sequencing step) in species identification is to use available barcode data to design optimized primers for further analysis, such as high-resolution melting analysis (HRM). This study aimed to determine the effectiveness of the hybrid method Bar-HRM (DNA barcoding combined with HRM) to identify species that share similar external morphological features, rather than conduct traditional taxonomic identification that require major parts (leaf, flower, fruit) of the specimens. The specimens used for testing were those, which could not be identified at the species level and could either be Uvaria longipes or Uvaria wrayias, indicated by morphological identification. Primer pairs derived from chloroplast regions (matK, psbA-trnH, rbcL, and trnL) were used in the Bar-HRM. The results obtained from psbA-trnH primers were good enough to help in identifying the specimen while the rest were not. Bar-HRM analysis was proven to be a fast and cost-effective method for plant species identification.

  5. High resolution melting: improvements in the genetic diagnosis of hypertrophic cardiomyopathy in a Portuguese cohort

    Directory of Open Access Journals (Sweden)

    Santos Susana

    2012-03-01

    Full Text Available Abstract Background Hypertrophic Cardiomyopathy (HCM is a complex myocardial disorder with a recognized genetic heterogeneity. The elevated number of genes and mutations involved in HCM limits a gene-based diagnosis that should be considered of most importance for basic research and clinical medicine. Methodology In this report, we evaluated High Resolution Melting (HRM robustness, regarding HCM genetic testing, by means of analyzing 28 HCM-associated genes, including the most frequent 4 HCM-associated sarcomere genes, as well as 24 genes with lower reported HCM-phenotype association. We analyzed 80 Portuguese individuals with clinical phenotype of HCM allowing simultaneously a better characterization of this disease in the Portuguese population. Results HRM technology allowed us to identify 60 mutated alleles in 72 HCM patients: 49 missense mutations, 3 nonsense mutations, one 1-bp deletion, one 5-bp deletion, one in frame 3-bp deletion, one insertion/deletion, 3 splice mutations, one 5'UTR mutation in MYH7, MYBPC3, TNNT2, TNNI3, CSRP3, MYH6 and MYL2 genes. Significantly 22 are novel gene mutations. Conclusions HRM was proven to be a technique with high sensitivity and a low false positive ratio allowing a rapid, innovative and low cost genotyping of HCM. In a short return, HRM as a gene scanning technique could be a cost-effective gene-based diagnosis for an accurate HCM genetic diagnosis and hopefully providing new insights into genotype/phenotype correlations.

  6. Detection of somatic mutations by high-resolution DNA melting (HRM analysis in multiple cancers.

    Directory of Open Access Journals (Sweden)

    Jesus Gonzalez-Bosquet

    Full Text Available Identification of somatic mutations in cancer is a major goal for understanding and monitoring the events related to cancer initiation and progression. High resolution melting (HRM curve analysis represents a fast, post-PCR high-throughput method for scanning somatic sequence alterations in target genes. The aim of this study was to assess the sensitivity and specificity of HRM analysis for tumor mutation screening in a range of tumor samples, which included 216 frozen pediatric small rounded blue-cell tumors as well as 180 paraffin-embedded tumors from breast, endometrial and ovarian cancers (60 of each. HRM analysis was performed in exons of the following candidate genes known to harbor established commonly observed mutations: PIK3CA, ERBB2, KRAS, TP53, EGFR, BRAF, GATA3, and FGFR3. Bi-directional sequencing analysis was used to determine the accuracy of the HRM analysis. For the 39 mutations observed in frozen samples, the sensitivity and specificity of HRM analysis were 97% and 87%, respectively. There were 67 mutation/variants in the paraffin-embedded samples, and the sensitivity and specificity for the HRM analysis were 88% and 80%, respectively. Paraffin-embedded samples require higher quantity of purified DNA for high performance. In summary, HRM analysis is a promising moderate-throughput screening test for mutations among known candidate genomic regions. Although the overall accuracy appears to be better in frozen specimens, somatic alterations were detected in DNA extracted from paraffin-embedded samples.

  7. High resolution melting analysis: a rapid and accurate method to detect CALR mutations.

    Directory of Open Access Journals (Sweden)

    Cristina Bilbao-Sieyro

    Full Text Available The recent discovery of CALR mutations in essential thrombocythemia (ET and primary myelofibrosis (PMF patients without JAK2/MPL mutations has emerged as a relevant finding for the molecular diagnosis of these myeloproliferative neoplasms (MPN. We tested the feasibility of high-resolution melting (HRM as a screening method for rapid detection of CALR mutations.CALR was studied in wild-type JAK2/MPL patients including 34 ET, 21 persistent thrombocytosis suggestive of MPN and 98 suspected secondary thrombocytosis. CALR mutation analysis was performed through HRM and Sanger sequencing. We compared clinical features of CALR-mutated versus 45 JAK2/MPL-mutated subjects in ET.Nineteen samples showed distinct HRM patterns from wild-type. Of them, 18 were mutations and one a polymorphism as confirmed by direct sequencing. CALR mutations were present in 44% of ET (15/34, 14% of persistent thrombocytosis suggestive of MPN (3/21 and none of the secondary thrombocytosis (0/98. Of the 18 mutants, 9 were 52 bp deletions, 8 were 5 bp insertions and other was a complex mutation with insertion/deletion. No mutations were found after sequencing analysis of 45 samples displaying wild-type HRM curves. HRM technique was reproducible, no false positive or negative were detected and the limit of detection was of 3%.This study establishes a sensitive, reliable and rapid HRM method to screen for the presence of CALR mutations.

  8. High-resolution melting analysis for prenatal diagnosis of beta-thalassemia in northern Thailand.

    Science.gov (United States)

    Charoenkwan, Pimlak; Sirichotiyakul, Supatra; Phusua, Arunee; Suanta, Sudjai; Fanhchaksai, Kanda; Sae-Tung, Rattika; Sanguansermsri, Torpong

    2017-12-01

    High-resolution melting (HRM) analysis is a rapid mutation analysis which assesses the pattern of reduction of fluorescence signal after subjecting the amplified PCR product with saturated fluorescence dye to an increasing temperature. We used HRM analysis for prenatal diagnosis of beta-thalassemia disease in northern Thailand. Five PCR-HRM protocols were used to detect point mutations in five different segments of the beta-globin gene, and one protocol to detect the 3.4 kb beta-globin deletion. We sought to characterize the mutations in carriers and to enable prenatal diagnosis in 126 couples at risk of having a fetus with beta-thalassemia disease. The protocols identified 18 common mutations causing beta-thalassemia, including the rare codon 132 (A-T) mutation. Each mutation showed a specific HRM pattern and all results were in concordance with those from direct DNA sequencing or gap-PCR methods. In cases of beta-thalassemia disease resulting from homozygosity for a mutation or compound heterozygosity for two mutations on the same amplified segment, the HRM patterns were different to those of a single mutation and were specific for each combination. HRM analysis is a simple and useful method for mutation identification in beta-thalassemia carriers and prenatal diagnosis of beta-thalassemia in northern Thailand.

  9. Traceability of PDO Olive Oil “Terra di Bari” Using High Resolution Melting

    Directory of Open Access Journals (Sweden)

    Cinzia Montemurro

    2015-01-01

    Full Text Available The aim of the research was to verify the applicability of microsatellite (SSR markers in High Resolution Melting (HRM analysis for the identification of the olive cultivars used in the “Terra di Bari” PDO extra virgin olive oil. A panel of nine cultivars, widespread in Apulia region, was tested with seventeen SSR primer pairs and the PCR products were at first analysed with a Genetic Analyzer automatic sequencer. An identification key was obtained for the nine cultivars, which showed an unambiguous discrimination among the varieties constituting the “Terra di Bari” PDO extra virgin olive oil: Cima di Bitonto, Coratina, and Ogliarola. Subsequently, an SSR based method was set up with the DCA18 marker, coupled with HRM analysis for the distinction of the Terra di Bari olive oil from non-Terra di Bari olive oil using different mixtures. Thus, this analysis enabled the distinction and identification of the PDO mixtures. Hence, this assay provided a flexible, cost-effective, and closed-tube microsatellite genotyping method, well suited to varietal identification and authentication analysis in olive oil.

  10. Application of high-resolution melting analysis for authenticity testing of valuable Dendrobium commercial products.

    Science.gov (United States)

    Dong, Xiaoman; Jiang, Chao; Yuan, Yuan; Peng, Daiyin; Luo, Yuqin; Zhao, Yuyang; Huang, Luqi

    2018-01-01

    The accurate identification of botanical origin in commercial products is important to ensure food authenticity and safety for consumers. The Dendrobium species have long been commercialised as functional food supplements and herbal medicines in Asia. Three valuable Dendrobium species, namely Dendrobium officinale, D. huoshanense and D. moniliforme, are often mutually adulterated in trade products in pursuit of higher profit. In this paper, a rapid and reliable semi-quantitative method for identifying the botanical origin of Dendrobium products in terminal markets was developed using high-resolution melting (HRM) analysis with specific primer pairs to target the trnL-F region. The HRM analysis method detected amounts of D. moniliforme adulterants as low as 1% in D. huoshanense or D. officinale products. The results have demonstrated that HRM analysis is a fast and effective tool for the differentiation of these Dendrobium species both for their authenticity as well as for the semi-quantitative determination of the purity of their processed products. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  11. Rapid discrimination of Isaria javanica and Isaria poprawskii from Isaria spp. using high resolution DNA melting assays

    Science.gov (United States)

    The current study evaluates the potential of using high resolution DNA melting assays to discriminate species in the genus, Isaria. The study utilizes a previously identified 103 base pair PCR amplicon, which was reported to be selective for Isaria fumosorosea. Our study finds the amplicon selective...

  12. Rapid Determination of Lymphogranuloma Venereum Serovars of Chlamydia trachomatis by Quantitative High-Resolution Melt Analysis (HRMA)

    Science.gov (United States)

    Stevens, Matthew P.; Garland, Suzanne M.; Zaia, Angelo M.; Tabrizi, Sepehr N.

    2012-01-01

    A quantitative high-resolution melt analysis assay was developed to differentiate lymphogranuloma venereum-causing serovars of Chlamydia trachomatis (L1 to L3) from other C. trachomatis serovars (D to K). The detection limit of this assay is approximately 10 copies per reaction, comparable to the limits of other quantitative-PCR-based methods. PMID:22933594

  13. ‘‘Blind'' mapping of genic DNA sequence polymorphisms in Lolium perenne L. by high resolution melting curve analysis

    DEFF Research Database (Denmark)

    Studer, Bruno; Jensen, Louise Bach; Fiil, Alice

    2009-01-01

    High resolution melting curve analysis (HRM) measures dissociation of double stranded DNA of a PCR product amplified in the presence of a saturating fluorescence dye. Recently, HRM proved successful to genotype DNA sequence polymorphisms such as SSRs and SNPs based on the shape of the melting...... curves. In this study, HRM was used for simultaneous screening and genotyping of genic DNA sequence polymorphisms identified in the Lolium perenne F2 mapping population VrnA. Melting profiles of PCR products amplified from previously published gene loci and from a novel gene putatively involved...

  14. Simplifying the detection of MUTYH mutations by high resolution melting analysis

    International Nuclear Information System (INIS)

    López-Villar, Isabel; Martínez-López, Joaquín; Ayala, Rosa; Wesselink, Jan; Morillas, Juan Diego; López, Elena; Marín, José Carlos; Díaz-Tasende, José; González, Sara; Robles, Luis

    2010-01-01

    MUTYH-associated polyposis (MAP) is a disorder caused by bi-allelic germline MUTYH mutation, characterized by multiple colorectal adenomas. In order to identify mutations in MUTYH gene we applied High Resolution Melting (HRM) genotyping. HRM analysis is extensively employed as a scanning method for the detection of heterozygous mutations. Therefore, we applied HRM to show effectiveness in detecting homozygous mutations for these clinically important and frequent patients. In this study, we analyzed phenotype and genotype data from 82 patients, with multiple (>= 10) synchronous (19/82) or metachronous (63/82) adenomas and negative APC study (except one case). Analysis was performed by HRM-PCR and direct sequencing, in order to identify mutations in MUTYH exons 7, 12 and 13, where the most prevalent mutations are located. In monoallelic mutation carriers, we evaluated entire MUTYH gene in search of another possible alteration. HRM-PCR was performed with strict conditions in several rounds: the first one to discriminate the heteroduplex patterns and homoduplex patterns and the next ones, in order to refine and confirm parameters. The genotypes obtained were correlated to phenotypic features (number of adenomas (synchronous or metachronous), colorectal cancer (CRC) and family history). MUTYH germline mutations were found in 15.8% (13/82) of patients. The hot spots, Y179C (exon 7) and G396D (exon 13), were readily identified and other mutations were also detected. Each mutation had a reproducible melting profile by HRM, both heterozygous mutations and homozygous mutations. In our study of 82 patients, biallelic mutation is associated with being a carrier of ≥10 synchronous polyps (p = 0.05) and there is no association between biallelic mutation and CRC (p = 0.39) nor family history (p = 0.63). G338H non-pathogenic polymorphism (exon 12) was found in 23.1% (19/82) of patients. In all cases there was concordance between HRM (first and subsequent rounds) and sequencing

  15. Evaluation of High Resolution Melting for MTHFR C677T Genotyping in Congenital Heart Disease.

    Directory of Open Access Journals (Sweden)

    Ying Wang

    Full Text Available High resolution melting (HRM is a simple, flexible and low-cost mutation screening technique. The methylenetetrahydrofolate reductase (MTHFR gene encoding a critical enzyme, potentially affects susceptibility to some congenital defects like congenital heart disease (CHD. We evaluate the performance of HRM for genotyping of the MTHFR gene C677T locus in CHD cases and healthy controls of Chinese Han population.A total of 315 blood samples from 147 CHD patients (male72, female 75 and 168 healthy controls (male 92, female 76 were enrolled in the study. HRM was utilized to genotype MTHFR C677T locus of all the samples. The results were compared to that of PCR-RFLP and Sanger sequencing. The association of the MTHFR C677T genotypes and the risk of CHD was analyzed using odds ratio with their 95% confidence interval (CIs from unconditional logistic regression.All the samples were successfully genotyped by HRM within 1 hour and 30 minutes while at least 6 hours were needed for PCR-RFLP and sequencing. The genotypes of MTHFR C677T CC, CT, and TT were 9.52%, 49.66%, and 40.82% in CHD group but 29.17%, 50% and 20.83% in control group, which were identical using both methods of HRM and PCR-RFLP, demonstrating the sensitivity and specificity of HRM were all 100%.MTHFR C677T is a potential risk factor for CHD in our local residents of Shandong province in China. HRM is a fast, sensitive, specific and reliable method for clinical application of genotyping.

  16. Molecular identification of broomrape species from a single seed by High Resolution Melting analysis

    Directory of Open Access Journals (Sweden)

    Mathieu Rolland

    2016-12-01

    Full Text Available Broomrapes are holoparasitic plants spreading through seeds. Each plant produces hundreds of thousands of seeds which remain viable in the soils for decades. To limit their spread, drastic measures are being taken and the contamination of a commercial seed lot by a single broomrape seed can lead to its rejection. Considering that broomrapes species identification from a single seed is extremely difficult even for trained botanists and that among all the described species, only a few are really noxious for the crops, numerous seed lots are rejected because of the contamination by seeds of non-noxious broomrape species. The aim of this study was to develop and evaluate a High Resolution Melting assay identifying the eight most noxious and common broomrape species (P. aegyptiaca, O. cernua, O. crenata, O. cumana, O. foetida, O. hederae, O. minor, and P. ramosa from a single seed. Based on trnL and rbcL plastidial genes amplification, the designed assay successfully identifies O. cumana, O. cernua, O. crenata, O. minor, O. hederae, and O. foetida; P. ramosa and P. aegyptiaca can be differentiated from other species but not from each other. Tested on 50 seed lots, obtained results perfectly matched identifications performed by sequencing. Through the analysis of common seed lots by different analysts, the reproducibility of the assay was evaluated at 90 %. Despite an original sample preparation process it was not possible to extract enough DNA from some seeds (10% of the samples. The described assay fulfils its objectives and allows an accurate identification of the targeted broomrape species. It can be used to identify contaminants in commercial seed lots or for any other purpose. The assay might be extended to vegetative material.

  17. High-resolution melting curve analysis for rapid detection of mutations in a Medaka TILLING library

    Directory of Open Access Journals (Sweden)

    Deguchi Tomonori

    2010-09-01

    Full Text Available Abstract Background During the last two decades, DNA sequencing has led to the identification of numerous genes in key species; however, in most cases, their functions are still unknown. In this situation, reverse genetics is the most suitable method to assign function to a gene. TILLING (Targeting Induced Local Lesions IN Genomes is a reverse-genetic strategy that combines random chemical mutagenesis with high-throughput discovery of the induced mutations in target genes. The method has been applied to a variety of plant and animal species. Screening of the induced mutations is the most important step in TILLING. Currently, direct sequencing or nuclease-mediated screening of heteroduplexes is widely used for detection of mutations in TILLING. Both methods are useful, but the costs are substantial and turnaround times are relatively long. Thus, there is a need for an alternative method that is of higher throughput and more cost effective. Results In this study, we developed a high resolution melting (HRM assay and evaluated its effectiveness for screening ENU-induced mutations in a medaka TILLING library. We had previously screened mutations in the p53 gene by direct sequencing. Therefore, we first tested the efficiency of the HRM assay by screening mutations in p53, which indicated that the HRM assay is as useful as direct sequencing. Next, we screened mutations in the atr and atm genes with the HRM assay. Nonsense mutations were identified in each gene, and the phenotypes of these nonsense mutants confirmed their loss-of-function nature. Conclusions These results demonstrate that the HRM assay is useful for screening mutations in TILLING. Furthermore, the phenotype of the obtained mutants indicates that medaka is an excellent animal model for investigating genome stability and gene function, especially when combined with TILLING.

  18. Transmission and selection of macrolide resistant Mycoplasma genitalium infections detected by rapid high resolution melt analysis.

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    Jimmy Twin

    Full Text Available BACKGROUND: Mycoplasma genitalium (MG causes urethritis, cervicitis and pelvic inflammatory disease. The MG treatment failure rate using 1 g azithromycin at an Australian Sexual Health clinic in 2007-9 was 31% (95%CI 23-40%. We developed a rapid high resolution melt analysis (HRMA assay targeting resistance mutations in the MG 23S rRNA gene, and validated it against DNA sequencing by examining pre- and post-treatment archived samples from MG-infected patients. METHODOLOGY/PRINCIPAL FINDINGS: Available MG-positive pre-treatment (n = 82 and post-treatment samples from individuals with clinical treatment failure (n = 20 were screened for 23S rRNA gene mutations. Sixteen (20% pre-treatment samples possessed resistance mutations (A2058G, A2059G, A2059C, which were significantly more common in patients with symptomatic azithromycin-treatment failure (12/26; 44% than in those clinically cured (4/56; 7%, p<0.001. All 20 patients experiencing azithromycin-failure had detectable mutations in their post-treatment samples. In 9 of these cases, the same mutational types were present in both pre- and post-treatment samples indicating transmitted resistance, whilst in 11 of these cases (55%, mutations were absent in pre-treatment samples indicating likely selection of resistant isolates have occurred. HRMA was able to detect all mutational changes determined in this study by DNA sequencing. An additional HRMA assay incorporating an unlabelled probe was also developed to detect type 4 single-nucleotide polymorphisms found in other populations, with a slightly lower sensitivity of 90%. CONCLUSIONS/SIGNIFICANCE: Treatment failure is associated with the detection of macrolide resistance mutations, which appear to be almost equally due to selection of resistant isolates following exposure to 1 g azithromycin and pre-existing transmitted resistance. The application of a rapid molecular assay to detect resistance at the time of initial detection of infection allows

  19. Rapid Detection and Differentiation of Clonorchis sinensis and Opisthorchis viverrini Using Real-Time PCR and High Resolution Melting Analysis

    OpenAIRE

    Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

    2014-01-01

    Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA...

  20. Greenland iceberg melt variability from high-resolution satellite observations

    Directory of Open Access Journals (Sweden)

    E. M. Enderlin

    2018-02-01

    Full Text Available Iceberg discharge from the Greenland Ice Sheet accounts for up to half of the freshwater flux to surrounding fjords and ocean basins, yet the spatial distribution of iceberg meltwater fluxes is poorly understood. One of the primary limitations for mapping iceberg meltwater fluxes, and changes over time, is the dearth of iceberg submarine melt rate estimates. Here we use a remote sensing approach to estimate submarine melt rates during 2011–2016 for 637 icebergs discharged from seven marine-terminating glaciers fringing the Greenland Ice Sheet. We find that spatial variations in iceberg melt rates generally follow expected patterns based on hydrographic observations, including a decrease in melt rate with latitude and an increase in melt rate with iceberg draft. However, we find no longitudinal variations in melt rates within individual fjords. We do not resolve coherent seasonal to interannual patterns in melt rates across all study sites, though we attribute a 4-fold melt rate increase from March to April 2011 near Jakobshavn Isbræ to fjord circulation changes induced by the seasonal onset of iceberg calving. Overall, our results suggest that remotely sensed iceberg melt rates can be used to characterize spatial and temporal variations in oceanic forcing near often inaccessible marine-terminating glaciers.

  1. Using high resolution tritium profiles to quantify the effects of melt on two Spitsbergen ice cores

    NARCIS (Netherlands)

    van der Wel, L.G.; Streurman, H.J.; Isaksson, E.; Helsen, M.M.; van de Wal, R.S.W.; Martma, T.; Pohjola, V.A.; Moore, J.C.; Meijer, H.A.J.

    2011-01-01

    Ice cores from small ice caps provide valuable climatic information, additional to that of Greenland and Antarctica. However, their integrity is usually compromised by summer meltwater percolation. To determine to what extent this can affect such ice cores, we performed high-resolution tritium

  2. High-resolution melt-curve analysis of random-amplified-polymorphic-DNA markers, for the characterisation of pathogenic Leptospira

    DEFF Research Database (Denmark)

    Tulsiani, Suhella; Craig, S B; Graham, G C

    2010-01-01

    A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used...... typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars....... These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation...

  3. Limited copy number-high resolution melting (LCN-HRM) enables the detection and identification by sequencing of low level mutations in cancer biopsies.

    Science.gov (United States)

    Do, Hongdo; Dobrovic, Alexander

    2009-10-08

    Mutation detection in clinical tumour samples is challenging when the proportion of tumour cells, and thus mutant alleles, is low. The limited sensitivity of conventional sequencing necessitates the adoption of more sensitive approaches. High resolution melting (HRM) is more sensitive than sequencing but identification of the mutation is desirable, particularly when it is important to discriminate false positives due to PCR errors or template degradation from true mutations.We thus developed limited copy number - high resolution melting (LCN-HRM) which applies limiting dilution to HRM. Multiple replicate reactions with a limited number of target sequences per reaction allow low level mutations to be detected. The dilutions used (based on Ct values) are chosen such that mutations, if present, can be detected by the direct sequencing of amplicons with aberrant melting patterns. Using cell lines heterozygous for mutations, we found that the mutations were not readily detected when they comprised 10% of total alleles (20% tumour cells) by sequencing, whereas they were readily detectable at 5% total alleles by standard HRM. LCN-HRM allowed these mutations to be identified by direct sequencing of those positive reactions.LCN-HRM was then used to review formalin-fixed paraffin-embedded (FFPE) clinical samples showing discordant findings between sequencing and HRM for KRAS exon 2 and EGFR exons 19 and 21. Both true mutations present at low levels and sequence changes due to artefacts were detected by LCN-HRM. The use of high fidelity polymerases showed that the majority of the artefacts were derived from the damaged template rather than replication errors during amplification. LCN-HRM bridges the sensitivity gap between HRM and sequencing and is effective in distinguishing between artefacts and true mutations.

  4. High-resolution melting genotyping of Enterococcus faecium based on multilocus sequence typing derived single nucleotide polymorphisms.

    Directory of Open Access Journals (Sweden)

    Steven Y C Tong

    Full Text Available We have developed a single nucleotide polymorphism (SNP nucleated high-resolution melting (HRM technique to genotype Enterococcus faecium. Eight SNPs were derived from the E. faecium multilocus sequence typing (MLST database and amplified fragments containing these SNPs were interrogated by HRM. We tested the HRM genotyping scheme on 85 E. faecium bloodstream isolates and compared the results with MLST, pulsed-field gel electrophoresis (PFGE and an allele specific real-time PCR (AS kinetic PCR SNP typing method. In silico analysis based on predicted HRM curves according to the G+C content of each fragment for all 567 sequence types (STs in the MLST database together with empiric data from the 85 isolates demonstrated that HRM analysis resolves E. faecium into 231 "melting types" (MelTs and provides a Simpson's Index of Diversity (D of 0.991 with respect to MLST. This is a significant improvement on the AS kinetic PCR SNP typing scheme that resolves 61 SNP types with D of 0.95. The MelTs were concordant with the known ST of the isolates. For the 85 isolates, there were 13 PFGE patterns, 17 STs, 14 MelTs and eight SNP types. There was excellent concordance between PFGE, MLST and MelTs with Adjusted Rand Indices of PFGE to MelT 0.936 and ST to MelT 0.973. In conclusion, this HRM based method appears rapid and reproducible. The results are concordant with MLST and the MLST based population structure.

  5. Identification of Brucella spp. isolated from human brucellosis in Malaysia using high-resolution melt (HRM) analysis.

    Science.gov (United States)

    Mohamed Zahidi, Jama'ayah; Bee Yong, Tay; Hashim, Rohaidah; Mohd Noor, Azura; Hamzah, Siti Hawa; Ahmad, Norazah

    2015-04-01

    Molecular approaches have been investigated to overcome difficulties in identification and differentiation of Brucella spp. using conventional phenotypic methods. In this study, high-resolution melt (HRM) analysis was used for rapid identification and differentiation of members of Brucella genus. A total of 41 Brucella spp. isolates from human brucellosis were subjected to HRM analysis using 4 sets of primers, which identified 40 isolates as Brucella melitensis and 1 as Brucella canis. The technique utilized low DNA concentration and was highly reproducible. The assay is shown to be a useful diagnostic tool, which can rapidly differentiate Brucella up to species level. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Quantifying the Evolution of Melt Ponds in the Marginal Ice Zone Using High Resolution Optical Imagery and Neural Networks

    Science.gov (United States)

    Ortiz, M.; Pinales, J. C.; Graber, H. C.; Wilkinson, J.; Lund, B.

    2016-02-01

    Melt ponds on sea ice play a significant and complex role on the thermodynamics in the Marginal Ice Zone (MIZ). Ponding reduces the sea ice's ability to reflect sunlight, and in consequence, exacerbates the albedo positive feedback cycle. In order to understand how melt ponds work and their effect on the heat uptake of sea ice, we must quantify ponds through their seasonal evolution first. A semi-supervised neural network three-class learning scheme using a gradient descent with momentum and adaptive learning rate backpropagation function is applied to classify melt ponds/melt areas in the Beaufort Sea region. The network uses high resolution panchromatic satellite images from the MEDEA program, which are collocated with autonomous platform arrays from the Marginal Ice Zone Program, including ice mass-balance buoys, arctic weather stations and wave buoys. The goal of the study is to capture the spatial variation of melt onset and freeze-up of the ponds within the MIZ, and gather ponding statistics such as size and concentration. The innovation of this work comes from training the neural network as the melt ponds evolve over time; making the machine learning algorithm time-dependent, which has not been previously done. We will achieve this by analyzing the image histograms through quantification of the minima and maxima intensity changes as well as linking textural variation information of the imagery. We will compare the evolution of the melt ponds against several different array sites on the sea ice to explore if there are spatial differences among the separated platforms in the MIZ.

  7. Species identification in meat products: A new screening method based on high resolution melting analysis of cyt b gene.

    Science.gov (United States)

    Lopez-Oceja, A; Nuñez, C; Baeta, M; Gamarra, D; de Pancorbo, M M

    2017-12-15

    Meat adulteration by substitution with lower value products and/or mislabeling involves economic, health, quality and socio-religious issues. Therefore, identification and traceability of meat species has become an important subject to detect possible fraudulent practices. In the present study the development of a high resolution melt (HRM) screening method for the identification of eight common meat species is reported. Samples from Bos taurus, Ovis aries, Sus scrofa domestica, Equus caballus, Oryctolagus cuniculus, Gallus gallus domesticus, Meleagris gallopavo and Coturnix coturnix were analyzed through the amplification of a 148 bp fragment from the cyt b gene with a universal primer pair in HRM analyses. Melting profiles from each species, as well as from several DNA mixtures of these species and blind samples, allowed a successful species differentiation. The results demonstrated that the HRM method here proposed is a fast, reliable, and low-cost screening technique. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Identification case of evidence in timber tracing of Pinus radiate, using high-resolution melting (HRM) analysis.

    Science.gov (United States)

    Solano, Jaime; Anabalón, Leonardo; Encina, Francisco

    2016-03-01

    Fast, accurate detection of plant species and their hybrids using molecular tools will facilitate assessment and monitoring of timber tracing evidence. In this study the origin of unknown pine samples is determined for a case of timber theft in the region of Araucania southern Chile. We evaluate the utility of the trnL marker region for species identification applied to pine wood based on High Resolution Melting. This efficient tracing methods can be incorporated into forestry applications such as certification of origin. The object of this work was genotype identification using high-resolution melting (HRM) and trnL approaches for Pinus radiata (Don) in timber tracing evidence. Our results indicate that trnL is a very sensitive marker for delimiting species and HRM analysis was used successfully for genotyping Pinus samples for timber tracing purposes. Genotyping samples by HRM analysis with the trnL1 approach allowed us to differentiate two wood samples from the Pinaceae family: Pinus radiata (Don) and Pseudotsuga menziesii (Mirb.) Franco. The same approach with Pinus trnL wood was not able to discriminate between samples of Pinus radiata, indicating that the samples were genetically indistinguishable, possibly because they have the same genotype at this locus. Timber tracing with HRM analysis is expected to contribute to future forest certification schemes, control of illegal trading, and molecular traceability of Pinus spp. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  9. Identification of TNIP1 Polymorphisms by High Resolution Melting Analysis with Unlabelled Probe: Association with Systemic Lupus Erythematosus

    Directory of Open Access Journals (Sweden)

    Jie Zhang

    2012-01-01

    Full Text Available Background. TNFα-induced protein 3 (TNFAIP3 interacting with protein 1 (TNIP1 acts as a negative regulator of NF-κB and plays an important role in maintaining the homeostasis of immune system. A recent genome-wide association study (GWAS showed that the polymorphism of TNIP1 was associated with the disease risk of SLE in Caucasian. In this study, we investigated whether the association of TNIP1 with SLE was replicated in Chinese population. Methods. The association of TNIP1 SNP rs7708392 (G/C was determined by high resolution melting (HRM analysis with unlabeled probe in 285 SLE patients and 336 healthy controls. Results. A new SNP rs79937737 located on 5 bp upstream of rs7708392 was discovered during the HRM analysis. No association of rs7708392 or rs79937737 with the disease risk of SLE was found. Furthermore, rs7708392 and rs79937737 were in weak linkage disequilibrium (LD. Hypotypes analysis of the two SNPs also showed no association with SLE in Chinese population. Conclusions. High resolution melting analysis with unlabeled probes proves to be a powerful and efficient genotyping method for identifying and screening SNPs. No association of rs7708392 or rs79937737 with the disease risk of SLE was observed in Chinese population.

  10. High resolution laser micro sintering / melting using q-switched and high brilliant laser radiation

    Science.gov (United States)

    Exner, H.; Streek, A.

    2015-03-01

    Since the discovery of selective laser sintering/melting, numerous modifications have been made to upgrade or customize this technology for industrial purposes. Laser micro sintering (LMS) is one of those modifications: Powders with particles in the range of a few micrometers are used to obtain products with highly resolved structures. Pulses of a q-switched laser had been considered necessary in order to generate sinter layers from the micrometer scaled metal powders. LMS has been applied with powders from metals as well as from ceramic and cermet feedstock's to generate micro parts. Recent technological progress and the application of high brilliant continuous laser radiation have now allowed an efficient laser sintering/melting of micrometer scaled metal powders. Thereby it is remarkable that thin sinter layers are generated using high continuous laser power. The principles of the process, the state of the art in LMS concerning its advantages and limitations and furthermore the latest results of the recent development of this technology will be presented. Laser Micro Sintering / Laser Micro Melting (LMM) offer a vision for a new dimension of additive fabrication of miniature and precise parts also with application potential in all engineering fields.

  11. Identification of the GST-T1 and GST-M1 null genotypes using high resolution melting analysis.

    Science.gov (United States)

    Drobná, Zuzana; Del Razo, Luz Maria; Garcia-Vargas, Gonzalo; Sánchez-Ramírez, Blanca; González-Horta, Carmen; Ballinas-Casarrubias, Lourdes; Loomis, Dana; Stýblo, Miroslav

    2012-01-13

    Glutathione S-transferases, including GST-T1 and GST-M1, are known to be involved in the phase II detoxification pathways for xenobiotics as well as in the metabolism of endogenous compounds. Polymorphisms in these genes have been linked to an increased susceptibility to carcinogenesis and associated with risk factors that predispose to certain inflammatory diseases. In addition, GST-T1 and GST-M1 null genotypes have been shown to be responsible for interindividual variations in the metabolism of arsenic, a known human carcinogen. To assess the specific GST genotypes in the Mexican population chronically exposed to arsenic, we have developed a multiplex High Resolution Melting PCR (HRM-PCR) analysis using a LightCycler480 instrument. This method is based on analysis of the PCR product melting curve that discriminates PCR products according to their lengths and base sequences. Three pairs of primers that specifically recognize GST-T1, GST-M1, and β-globin, an internal control, to produce amplicons of different length were designed and combined with LightCycler480 High Resolution Melting Master Mix containing ResoLight, a completely saturating DNA dye. Data collected from melting curve analysis were evaluated using LightCycler480 software to determine specific melting temperatures of individual melting curves representing target genes. Using this newly developed multiplex HRM-PCR analysis, we evaluated GST-T1 and GST-M1 genotypes in 504 DNA samples isolated from the blood of individuals residing in Zimapan, Lagunera, and Chihuahua regions in Mexico. We found that the Zimapan and Lagunera populations have similar GST-T1 and GST-M1 genotype frequencies which differ from those of the Chihuahua population. In addition, 14 individuals have been identified as carriers of the double null genotype, i.e., null genotypes in both GST-T1 and GST-M1 genes. Although this procedure does not distinguish between biallelic (+/+) and monoallelic (+/-) genotypes, it can be used in an

  12. High-Resolution Melting Curve Analysis of the 16S Ribosomal Gene to Detect and Identify Pathogenic and Saprophytic Leptospira Species in Colombian Isolates.

    Science.gov (United States)

    Peláez Sánchez, Ronald G; Quintero, Juan Álvaro López; Pereira, Martha María; Agudelo-Flórez, Piedad

    2017-05-01

    AbstractIt is important to identify the circulating Leptospira agent to enhance the performance of serodiagnostic tests by incorporating specific antigens of native species, develop vaccines that take into account the species/serovars circulating in different regions, and optimize prevention and control strategies. The objectives of this study were to develop a polymerase chain reaction (PCR)-high-resolution melting (HRM) assay for differentiating between species of the genus Leptospira and to verify its usefulness in identifying unknown samples to species level. A set of primers from the initial region of the 16S ribosomal gene was designed to detect and differentiate the 22 species of Leptospira . Eleven reference strains were used as controls to establish the reference species and differential melting curves. Twenty-five Colombian Leptospira isolates were studied to evaluate the usefulness of the PCR-HRM assay in identifying unknown samples to species level. This identification was confirmed by sequencing and phylogenetic analysis of the 16S ribosomal gene. Eleven Leptospira species were successfully identified, except for Leptospira meyeri / Leptospira yanagawae because the sequences were 100% identical. The 25 isolates from humans, animals, and environmental water sources were identified as Leptospira santarosai (twelve), Leptospira interrogans (nine), and L. meyeri / L. yanagawae (four). The species verification was 100% concordant between PCR-HRM and phylogenetic analysis of the 16S ribosomal gene. The PCR-HRM assay designed in this study is a useful tool for identifying Leptospira species from isolates.

  13. High-resolution melting analysis using unlabeled probe and amplicon scanning simultaneously detects several lactase persistence variants

    DEFF Research Database (Denmark)

    Janukonyté, Jurgita; Vestergaard, Else M; Ladefoged, Søren A

    2010-01-01

    Lactase persistence and thereby tolerance to lactose is a common trait in people of Northern European descent. It is linked to the LCT -13910C>T variant located in intron 13 of the MCM6 gene 13.9 kb upstream of the lactase (LCT) gene. In people of African and Middle Eastern descent, lactase...... persistence can be associated with other variants nearby the -13910C>T variant, limiting the use of the -13910C>T-based SNP analysis, e.g. TaqMan assays for the diagnosis of lactose intolerance. Using high-resolution melting analysis, we identified five samples that were heterozygous for the -13915T>G variant...... the -13910C>T and -13915T>G variants in addition to rarer variants surrounding the -13910 site. This new method may contribute to improve the diagnostic performance of the genetic analysis for lactose intolerance....

  14. A novel typing method for Listeria monocytogenes using high-resolution melting analysis (HRMA) of tandem repeat regions.

    Science.gov (United States)

    Ohshima, Chihiro; Takahashi, Hajime; Iwakawa, Ai; Kuda, Takashi; Kimura, Bon

    2017-07-17

    Listeria monocytogenes, which is responsible for causing food poisoning known as listeriosis, infects humans and animals. Widely distributed in the environment, this bacterium is known to contaminate food products after being transmitted to factories via raw materials. To minimize the contamination of products by food pathogens, it is critical to identify and eliminate factory entry routes and pathways for the causative bacteria. High resolution melting analysis (HRMA) is a method that takes advantage of differences in DNA sequences and PCR product lengths that are reflected by the disassociation temperature. Through our research, we have developed a multiple locus variable-number tandem repeat analysis (MLVA) using HRMA as a simple and rapid method to differentiate L. monocytogenes isolates. While evaluating our developed method, the ability of MLVA-HRMA, MLVA using capillary electrophoresis, and multilocus sequence typing (MLST) was compared for their ability to discriminate between strains. The MLVA-HRMA method displayed greater discriminatory ability than MLST and MLVA using capillary electrophoresis, suggesting that the variation in the number of repeat units, along with mutations within the DNA sequence, was accurately reflected by the melting curve of HRMA. Rather than relying on DNA sequence analysis or high-resolution electrophoresis, the MLVA-HRMA method employs the same process as PCR until the analysis step, suggesting a combination of speed and simplicity. The result of MLVA-HRMA method is able to be shared between different laboratories. There are high expectations that this method will be adopted for regular inspections at food processing facilities in the near future. Copyright © 2017. Published by Elsevier B.V.

  15. High Resolution Melting (HRM) for High-Throughput Genotyping—Limitations and Caveats in Practical Case Studies

    Science.gov (United States)

    Słomka, Marcin; Sobalska-Kwapis, Marta; Wachulec, Monika; Bartosz, Grzegorz

    2017-01-01

    High resolution melting (HRM) is a convenient method for gene scanning as well as genotyping of individual and multiple single nucleotide polymorphisms (SNPs). This rapid, simple, closed-tube, homogenous, and cost-efficient approach has the capacity for high specificity and sensitivity, while allowing easy transition to high-throughput scale. In this paper, we provide examples from our laboratory practice of some problematic issues which can affect the performance and data analysis of HRM results, especially with regard to reference curve-based targeted genotyping. We present those examples in order of the typical experimental workflow, and discuss the crucial significance of the respective experimental errors and limitations for the quality and analysis of results. The experimental details which have a decisive impact on correct execution of a HRM genotyping experiment include type and quality of DNA source material, reproducibility of isolation method and template DNA preparation, primer and amplicon design, automation-derived preparation and pipetting inconsistencies, as well as physical limitations in melting curve distinction for alternative variants and careful selection of samples for validation by sequencing. We provide a case-by-case analysis and discussion of actual problems we encountered and solutions that should be taken into account by researchers newly attempting HRM genotyping, especially in a high-throughput setup. PMID:29099791

  16. High Resolution Melting (HRM) for High-Throughput Genotyping-Limitations and Caveats in Practical Case Studies.

    Science.gov (United States)

    Słomka, Marcin; Sobalska-Kwapis, Marta; Wachulec, Monika; Bartosz, Grzegorz; Strapagiel, Dominik

    2017-11-03

    High resolution melting (HRM) is a convenient method for gene scanning as well as genotyping of individual and multiple single nucleotide polymorphisms (SNPs). This rapid, simple, closed-tube, homogenous, and cost-efficient approach has the capacity for high specificity and sensitivity, while allowing easy transition to high-throughput scale. In this paper, we provide examples from our laboratory practice of some problematic issues which can affect the performance and data analysis of HRM results, especially with regard to reference curve-based targeted genotyping. We present those examples in order of the typical experimental workflow, and discuss the crucial significance of the respective experimental errors and limitations for the quality and analysis of results. The experimental details which have a decisive impact on correct execution of a HRM genotyping experiment include type and quality of DNA source material, reproducibility of isolation method and template DNA preparation, primer and amplicon design, automation-derived preparation and pipetting inconsistencies, as well as physical limitations in melting curve distinction for alternative variants and careful selection of samples for validation by sequencing. We provide a case-by-case analysis and discussion of actual problems we encountered and solutions that should be taken into account by researchers newly attempting HRM genotyping, especially in a high-throughput setup.

  17. A high-resolution melting (HRM) assay for the differentiation between Israeli field and Neethling vaccine lumpy skin disease viruses.

    Science.gov (United States)

    Menasherow, Sophia; Erster, Oran; Rubinstein-Giuni, Marisol; Kovtunenko, Anita; Eyngor, Evgeny; Gelman, Boris; Khinich, Evgeny; Stram, Yehuda

    2016-06-01

    Lumpy skin disease (LSD) is a constant threat to the Middle East including the State of Israel. During vaccination programs it is essential for veterinary services and farmers to be able to distinguish between animals affected by the cattle-borne virulent viruses and vaccinated animals, subsequently affected by the vaccine strain. This study describes an improved high resolution-melting (HRM) test that exploits a 27 base pair (bp) fragment of the LSDV126 extracellular enveloped virion (EEV) gene that is present in field viruses but is absent from the Neethling vaccine strain. This difference leads to ∼0.5 °C melting point change in the HRM assay, when testing the quantitative PCR (qPCR) products generated from the virulent field viruses compared to the attenuated vaccine. By exploiting this difference, it could be shown using the newly developed HRM assay that virus isolated from vaccinated cattle that developed disease symptoms behave similarly to vaccine virus control, indicating that the vaccine virus can induce disease symptoms. This assay is not only in full agreement with the previously published PCR gradient and restriction fragment length polymorphism (RFLP) tests but it is faster with, fewer steps, cheaper and dependable. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. High Resolution Melting (HRM for High-Throughput Genotyping—Limitations and Caveats in Practical Case Studies

    Directory of Open Access Journals (Sweden)

    Marcin Słomka

    2017-11-01

    Full Text Available High resolution melting (HRM is a convenient method for gene scanning as well as genotyping of individual and multiple single nucleotide polymorphisms (SNPs. This rapid, simple, closed-tube, homogenous, and cost-efficient approach has the capacity for high specificity and sensitivity, while allowing easy transition to high-throughput scale. In this paper, we provide examples from our laboratory practice of some problematic issues which can affect the performance and data analysis of HRM results, especially with regard to reference curve-based targeted genotyping. We present those examples in order of the typical experimental workflow, and discuss the crucial significance of the respective experimental errors and limitations for the quality and analysis of results. The experimental details which have a decisive impact on correct execution of a HRM genotyping experiment include type and quality of DNA source material, reproducibility of isolation method and template DNA preparation, primer and amplicon design, automation-derived preparation and pipetting inconsistencies, as well as physical limitations in melting curve distinction for alternative variants and careful selection of samples for validation by sequencing. We provide a case-by-case analysis and discussion of actual problems we encountered and solutions that should be taken into account by researchers newly attempting HRM genotyping, especially in a high-throughput setup.

  19. Molecular differentiation of Opisthorchis viverrini and Clonorchis sinensis eggs by multiplex real-time PCR with high resolution melting analysis.

    Science.gov (United States)

    Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Laummaunwai, Porntip; Lulitanond, Viraphong; Doanh, Pham Ngoc; Maleewong, Wanchai

    2013-12-01

    Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at 82.4±0.09℃ and 85.9±0.08℃ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

  20. The use of high resolution melting analysis to detect Fabry mutations in heterozygous females via dry bloodspots.

    Science.gov (United States)

    Tai, Chang-Long; Liu, Mei-Ying; Yu, Hsiao-Chi; Chiang, Chiang-Chuan; Chiang, Hung; Suen, Jeng-Hung; Kao, Shu-Min; Huang, Yu-Hsiu; Wu, Tina Jui-Ting; Yang, Chia-Feng; Tsai, Fang-Chih; Lin, Ching-Yuang; Chang, Jan-Gowth; Chen, Hong-Duo; Niu, Dau-Ming

    2012-02-18

    As an X-linked genetic disorder, Fabry disease was first thought to affect males only, and females were generally considered to be asymptomatic carriers. However, recent research suggests that female carriers of Fabry disease may still develop vital organ damage causing severe morbidity and mortality. In the previous newborn screening, from 299,007 newborns, we identified a total of 20 different Fabry mutations and 121 newborns with Fabry mutations. However, we found that most female carriers are not detected by enzyme assays. A streamlined method for high resolution melting (HRM) analysis was designed to screen for GLA gene mutations using a same PCR and melting program. Primer sets were designed to cover the 7 exons and the Chinese common intronic mutation, IVS4+919G>A of GLA gene. The HRM analysis was successful in identifying heterozygous and hemizygous patients with the 20 surveyed mutations. We were also successful in using this method to test dry blood spots of newborns afflicted with Fabry mutations without having to determine DNA concentration before PCR amplification. The results of this study show that HRM could be a reliable and sensitive method for use in the rapid screening of females for GLA mutations. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. The WAIS Melt Monitor: An automated ice core melting system for meltwater sample handling and the collection of high resolution microparticle size distribution data

    Science.gov (United States)

    Breton, D. J.; Koffman, B. G.; Kreutz, K. J.; Hamilton, G. S.

    2010-12-01

    Paleoclimate data are often extracted from ice cores by careful geochemical analysis of meltwater samples. The analysis of the microparticles found in ice cores can also yield unique clues about atmospheric dust loading and transport, dust provenance and past environmental conditions. Determination of microparticle concentration, size distribution and chemical makeup as a function of depth is especially difficult because the particle size measurement either consumes or contaminates the meltwater, preventing further geochemical analysis. Here we describe a microcontroller-based ice core melting system which allows the collection of separate microparticle and chemistry samples from the same depth intervals in the ice core, while logging and accurately depth-tagging real-time electrical conductivity and particle size distribution data. This system was designed specifically to support microparticle analysis of the WAIS Divide WDC06A deep ice core, but many of the subsystems are applicable to more general ice core melting operations. Major system components include: a rotary encoder to measure ice core melt displacement with 0.1 millimeter accuracy, a meltwater tracking system to assign core depths to conductivity, particle and sample vial data, an optical debubbler level control system to protect the Abakus laser particle counter from damage due to air bubbles, a Rabbit 3700 microcontroller which communicates with a host PC, collects encoder and optical sensor data and autonomously operates Gilson peristaltic pumps and fraction collectors to provide automatic sample handling, melt monitor control software operating on a standard PC allowing the user to control and view the status of the system, data logging software operating on the same PC to collect data from the melting, electrical conductivity and microparticle measurement systems. Because microparticle samples can easily be contaminated, we use optical air bubble sensors and high resolution ice core density

  2. High resolution melting curve analysis as a new tool for rapid identification of canine parvovirus type 2 strains.

    Science.gov (United States)

    Bingga, Gali; Liu, Zhicheng; Zhang, Jianfeng; Zhu, Yujun; Lin, Lifeng; Ding, Shuangyang; Guo, Pengju

    2014-01-01

    A high resolution melting (HRM) curve method was developed to identify canine parvovirus type 2 (CPV-2) strains by nested PCR. Two sets of primers, CPV-426F/426R and CPV-87R/87F, were designed that amplified a 52 bp and 53 bp product from the viral VP2 capsid gene. The region amplified by CPV-426F/426R included the A4062G and T4064A mutations in CPV-2a, CPV-2b and CPV-2c. The region amplified by CPV-87F/87R included the A3045T mutation in the vaccine strains of CPV-2 and CPV-2a, CPV-2b and CPV-2c. Faecal samples were obtained from 30 dogs that were CPV antigen-positive. The DNA was isolated from the faecal samples and PCR-amplified using the two sets of primers, and genotyped by HRM curve analysis. The PCR-HRM assay was able to distinguish single nucleotide polymorphisms between CPV-2a, CPV-2b and CPV-2c using CPV-426F/426R. CPV-2a was distinguished from CPV-2b and CPV-2c by differences in the melting temperature. CPV-2b and CPV-2c could be distinguished based on the shape of the melting curve after generating heteroduplexes using a CPV-2b reference sample. The vaccine strains of CPV-2 were identified using CPV-87F/87R. Conventional methods for genotyping CPV strains are labor intensive, expensive or time consuming; the present PCR-based HRM assay might be an attractive alternative. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. A high resolution melting (HRM) technology-based assay for cost-efficient clinical detection and genotyping of herpes simplex virus (HSV)-1 and HSV-2.

    Science.gov (United States)

    Lieveld, M; Carregosa, A; Benoy, I; Redzic, N; Berth, M; Vanden Broeck, D

    2017-10-01

    Genital herpes can be caused by two very similar viruses, herpes simplex virus (HSV)-1 or HSV-2. These two HSV types cannot be distinguished clinically, but genotyping is recommended in the first-episodes of genital herpes to guide counselling and management. Quantitative polymerase chain reaction (qPCR) is the preferred diagnostic method for HSV typing. However, commercial qPCR methods use expensive fluorescent labeled probes for detection. Furthermore, most low-cost methods are not able to differentiate between HSV-1 and -2. The aim of this study was to develop a high resolution melting (HRM) technology-based assay for sensitive HSV-1 and HSV-2 detection and genotyping. Using a panel of 46 clinical specimens, the performance of the HRM assay was compared to two commercial HSV tests: the HRM assay detected HSV in all 23 positive samples, with no false positive results (100% concordance with HSV I/II Real-TM assay). Additionally, the HRM assay correctly genotyped both HSV types in a subset of these clinical samples, as determined by the Realstar HSV PCR Kit. The HSV HRM assay provides a cost-effective alternative method to conventional more expensive assays and can be used in routine clinical specimens, in cases where it is particularly necessary to detect and distinguish HSV-1 from -2. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Development and assessment of multiplex high resolution melting assay as a tool for rapid single-tube identification of five Brucella species.

    Science.gov (United States)

    Gopaul, Krishna K; Sells, Jessica; Lee, Robin; Beckstrom-Sternberg, Stephen M; Foster, Jeffrey T; Whatmore, Adrian M

    2014-12-11

    The zoonosis brucellosis causes economically significant reproductive problems in livestock and potentially debilitating disease of humans. Although the causative agent, organisms from the genus Brucella, can be differentiated into a number of species based on phenotypic characteristics, there are also significant differences in genotype that are concordant with individual species. This paper describes the development of a five target multiplex assay to identify five terrestrial Brucella species using real-time polymerase chain reaction (PCR) and subsequent high resolution melt curve analysis. This technology offers a robust and cost effective alternative to previously described hydrolysis-probe Single Nucleotide Polymorphism (SNP)-based species defining assays. Through the use of Brucella whole genome sequencing five species defining SNPs were identified. Individual HRM assays were developed to these target these changes and, following optimisation of primer concentrations, it was possible to multiplex all five assays in a single tube. In a validation exercise using a panel of 135 Brucella strains of terrestrial and marine origin, it was possible to distinguish the five target species from the other species within this panel. The HRM multiplex offers a number of diagnostic advantages over previously described SNP-based typing approaches. Further, and uniquely for HRM, the successful multiplexing of five assays in a single tube allowing differentiation of five Brucella species in the diagnostic laboratory in a cost-effective and timely manner is described. However there are possible limitations to using this platform on DNA extractions direct from clinical material.

  5. Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

    Science.gov (United States)

    Ben Ali, Sina-Elisabeth; Madi, Zita Erika; Hochegger, Rupert; Quist, David; Prewein, Bernhard; Haslberger, Alexander G.; Brandes, Christian

    2014-01-01

    Genetic mutations must be avoided during the production and use of seeds. In the European Union (EU), Directive 2001/18/EC requires any DNA construct introduced via transformation to be stable. Establishing genetic stability is critical for the approval of genetically modified organisms (GMOs). In this study, genetic stability of two GMOs was examined using high resolution melting (HRM) analysis and real-time polymerase chain reaction (PCR) employing Scorpion primers for amplification. The genetic variability of the transgenic insert and that of the flanking regions in a single oilseed rape variety (GT73) and a stacked maize (MON88017 × MON810) was studied. The GT73 and the 5' region of MON810 showed no instabilities in the examined regions. However; two out of 100 analyzed samples carried a heterozygous point mutation in the 3' region of MON810 in the stacked variety. These results were verified by direct sequencing of the amplified PCR products as well as by sequencing of cloned PCR fragments. The occurrence of the mutation suggests that the 5' region is more suitable than the 3' region for the quantification of MON810. The identification of the single nucleotide polymorphism (SNP) in a stacked event is in contrast to the results of earlier studies of the same MON810 region in a single event where no DNA polymorphism was found. PMID:25365178

  6. Detection of Schistosoma mansoni and Schistosoma haematobium by Real-Time PCR with High Resolution Melting Analysis

    Directory of Open Access Journals (Sweden)

    Hany Sady

    2015-07-01

    Full Text Available The present study describes a real-time PCR approach with high resolution melting-curve (HRM assay developed for the detection and differentiation of Schistosoma mansoni and S. haematobium in fecal and urine samples collected from rural Yemen. The samples were screened by microscopy and PCR for the Schistosoma species infection. A pair of degenerate primers were designed targeting partial regions in the cytochrome oxidase subunit I (cox1 gene of S. mansoni and S. haematobium using real-time PCR-HRM assay. The overall prevalence of schistosomiasis was 31.8%; 23.8% of the participants were infected with S. haematobium and 9.3% were infected with S. mansoni. With regards to the intensity of infections, 22.1% and 77.9% of S. haematobium infections were of heavy and light intensities, respectively. Likewise, 8.1%, 40.5% and 51.4% of S. mansoni infections were of heavy, moderate and light intensities, respectively. The melting points were distinctive for S. mansoni and S. haematobium, categorized by peaks of 76.49 ± 0.25 °C and 75.43 ± 0.26 °C, respectively. HRM analysis showed high detection capability through the amplification of Schistosoma DNA with as low as 0.0001 ng/µL. Significant negative correlations were reported between the real-time PCR-HRM cycle threshold (Ct values and microscopic egg counts for both S. mansoni in stool and S. haematobium in urine (p < 0.01. In conclusion, this closed-tube HRM protocol provides a potentially powerful screening molecular tool for the detection of S. mansoni and S. haematobium. It is a simple, rapid, accurate, and cost-effective method. Hence, this method is a good alternative approach to probe-based PCR assays.

  7. Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis.

    Science.gov (United States)

    Hanson, Erin K; Ballantyne, Jack

    2013-01-01

    Positive identification of the nature of biological material present on evidentiary items can be crucial for understanding the circumstances surrounding a crime. However, traditional protein-based methods do not permit the identification of all body fluids and tissues, and thus molecular based strategies for the conclusive identification of all forensically relevant biological fluids and tissues need to be developed. Messenger RNA (mRNA) profiling is an example of such a molecular-based approach. Current mRNA body fluid identification assays involve capillary electrophoresis (CE) or quantitative RT-PCR (qRT-PCR) platforms, each with its own limitations. Both platforms require the use of expensive fluorescently labeled primers or probes. CE-based assays require separate amplification and detection steps thus increasing the analysis time. For qRT-PCR assays, only 3-4 markers can be included in a single reaction since each requires a different fluorescent dye. To simplify mRNA profiling assays, and reduce the time and cost of analysis, we have developed single- and multiplex body fluid High Resolution Melt (HRM) assays for the identification of common forensically relevant biological fluids and tissues. The incorporated biomarkers include IL19 (vaginal secretions), IL1F7 (skin), ALAS2 (blood), MMP10 (menstrual blood), HTN3 (saliva) and TGM4 (semen).  The HRM assays require only unlabeled PCR primers and a single saturating intercalating fluorescent dye (Eva Green). Each body-fluid-specific marker can easily be identified by the presence of a distinct melt peak. Usually, HRM assays are used to detect variants or isoforms for a single gene target. However, we have uniquely developed duplex and triplex HRM assays to permit the simultaneous detection of multiple targets per reaction. Here we describe the development and initial performance evaluation of the developed HRM assays. The results demonstrate the potential use of HRM assays for rapid, and relatively inexpensive

  8. Identification of six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay.

    Science.gov (United States)

    Hernández, Carolina; Alvarez, Catalina; González, Camila; Ayala, Martha Stella; León, Cielo Maritza; Ramírez, Juan David

    2014-11-14

    Leishmaniases are tropical zoonotic diseases, caused by parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. This eco-epidemiological complexity imposes a challenge to the detection of circulating parasite species, not only related to human cases but also infecting vectors and reservoirs. Currently, no harmonized methods have been deployed to discriminate the NW Leishmania species. Herein, we conducted a systematic and mechanistic High-Resolution Melting (HRM) assay targeted to HSP70 and ITS1. Specific primers were designed that coupled with a HRM analyses permitted to discriminate six NW Leishmania species. In order to validate the herein described algorithm, we included 35 natural isolates obtained from human cases, insect vectors and mammals. Our genotyping assay allowed the correct assignment of the six NW Leishmania species (L. mexicana, L. infantum (chagasi), L. amazonensis, L. panamensis, L. guyanensis and L. braziliensis) based on reference strains. When the algorithm was applied to a set of well-characterized strains by means of PCR-RFLP, MLEE and monoclonal antibodies (MA) we observed a tailored concordance between the HRM and PCR-RFLP/MLEE/MA (KI = 1.0). Additionally, we tested the limit of detection for the HRM method showing that this is able to detect at least 10 equivalent-parasites per mL. This is a rapid and reliable method to conduct molecular epidemiology and host-parasite association studies in endemic areas.

  9. Application of high-resolution DNA melting for genotyping in lepidopteran non-model species: Ostrinia furnacalis (Crambidae.

    Directory of Open Access Journals (Sweden)

    FengBo Li

    Full Text Available Development of an ideal marker system facilitates a better understanding of the genetic diversity in lepidopteran non-model organisms, which have abundant species, but relatively limited genomic resources. Single nucleotide polymorphisms (SNPs discovered within single-copy genes have proved to be desired markers, but SNP genotyping by current techniques remain laborious and expensive. High resolution melting (HRM curve analysis represents a simple, rapid and inexpensive genotyping method that is primarily confined to clinical and diagnostic studies. In this study, we evaluated the potential of HRM analysis for SNP genotyping in the lepidopteran non-model species Ostrinia furnacalis (Crambidae. Small amplicon and unlabeled probe assays were developed for the SNPs, which were identified in 30 females of O. furnacalis from 3 different populations by our direct sequencing. Both assays were then applied to genotype 90 unknown female DNA by prior mixing with known wild-type DNA. The genotyping results were compared with those that were obtained using bi-directional sequencing analysis. Our results demonstrated the efficiency and reliability of the HRM assays. HRM has the potential to provide simple, cost-effective genotyping assays and facilitates genotyping studies in any non-model lepidopteran species of interest.

  10. First report of Tasmanian sheep strain (G2) genotype isolated from Iranian goat using the high resolution melting (HRM) analysis.

    Science.gov (United States)

    Hosseini-Safa, Ahmad; Mohag Hegh, Mohammad Ali; Pestechian, Nader; Ganji, Maryam; Mohammadi, Rasoul; Mahmoudi Lamouki, Reza; Rostami-Nejad, Mohammad

    2016-12-01

    The present study was aimed to evaluate E. granulosus genotypes isolated from goats using HRM analysis in Isfahan province. Cystic echincoccosis, so-called hydatidosis, is widespread infection caused by the larval stage of Echinococcus granulosus . This is an important zoonotic disease worldwide, especially in the developing countries such as Iran. To date, molecular studies mainly based on the mitochondrial DNA sequences have identified distinct genotypes termed G1-G10 which can differ in some characteristics such as the growth and infectivity to different intermediate hosts or the survival rate in the definitive hosts that are important for the development of control strategies. From August to December 2014, 1341 goats were investigated and hydatid cysts were collected from the liver and lungs of 43 infected goats in Isfahan province abattoirs, Isfahan, Iran. Total genomic DNA was extracted from each sample, amplified for the presence of polymorphism of mitochondrial gene coding for cytochrome c oxidase subunit 1 (CO1), using high resolution melting curve (HRM) method. the results of HRM analysis using the sequence of CO1 gene for 43 Echinococcus granulosus isolates from goats showed 31, 2 and 10 isolates were identified as G1, G2, and G3 genotypes, respectively. G1 is the predominant genotype in the isolated goat samples in Isfahan province, and the presence of G2 strain was reported for the first time in goat in Iran.

  11. Validation of high-resolution DNA melting analysis for mutation scanning of the CDKL5 gene: identification of novel mutations.

    Science.gov (United States)

    Raymond, Laure; Diebold, Bertrand; Leroux, Céline; Maurey, Hélène; Drouin-Garraud, Valérie; Delahaye, Andre; Dulac, Olivier; Metreau, Julia; Melikishvili, Gia; Toutain, Annick; Rivier, François; Bahi-Buisson, Nadia; Bienvenu, Thierry

    2013-01-01

    Mutations in the cyclin-dependent kinase-like 5 gene (CDKL5) have been predominantly described in epileptic encephalopathies of female, including infantile spasms with Rett-like features. Up to now, detection of mutations in this gene was made by laborious, expensive and/or time consuming methods. Here, we decided to validate high-resolution melting analysis (HRMA) for mutation scanning of the CDKL5 gene. Firstly, using a large DNA bank consisting to 34 samples carrying different mutations and polymorphisms, we validated our analytical conditions to analyse the different exons and flanking intronic sequences of the CDKL5 gene by HRMA. Secondly, we screened CDKL5 by both HRMA and denaturing high performance liquid chromatography (dHPLC) in a cohort of 135 patients with early-onset seizures. Our results showed that point mutations and small insertions and deletions can be reliably detected by HRMA. Compared to dHPLC, HRMA profiles are more discriminated, thereby decreasing unnecessary sequencing. In this study, we identified eleven novel sequence variations including four pathogenic mutations (2.96% prevalence). HRMA appears cost-effective, easy to set up, highly sensitive, non-toxic and rapid for mutation screening, ideally suited for large genes with heterogeneous mutations located along the whole coding sequence, such as the CDKL5 gene. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. The potential of high resolution melting analysis (hrma) to streamline, facilitate and enrich routine diagnostics in medical microbiology.

    Science.gov (United States)

    Ruskova, Lenka; Raclavsky, Vladislav

    2011-09-01

    Routine medical microbiology diagnostics relies on conventional cultivation followed by phenotypic techniques for identification of pathogenic bacteria and fungi. This is not only due to tradition and economy but also because it provides pure culture needed for antibiotic susceptibility testing. This review focuses on the potential of High Resolution Melting Analysis (HRMA) of double-stranded DNA for future routine medical microbiology. Search of MEDLINE database for publications showing the advantages of HRMA in routine medical microbiology for identification, strain typing and further characterization of pathogenic bacteria and fungi in particular. The results show increasing numbers of newly-developed and more tailor-made assays in this field. For microbiologists unfamiliar with technical aspects of HRMA, we also provide insight into the technique from the perspective of microbial characterization. We can anticipate that the routine availability of HRMA in medical microbiology laboratories will provide a strong stimulus to this field. This is already envisioned by the growing number of medical microbiology applications published recently. The speed, power, convenience and cost effectiveness of this technology virtually predestine that it will advance genetic characterization of microbes and streamline, facilitate and enrich diagnostics in routine medical microbiology without interfering with the proven advantages of conventional cultivation.

  13. Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini using real-time PCR and high resolution melting analysis.

    Science.gov (United States)

    Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

    2014-01-01

    Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

  14. Rapid Detection and Differentiation of Clonorchis sinensis and Opisthorchis viverrini Using Real-Time PCR and High Resolution Melting Analysis

    Directory of Open Access Journals (Sweden)

    Xian-Quan Cai

    2014-01-01

    Full Text Available Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

  15. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L.) using high-resolution melting.

    Science.gov (United States)

    Zhang, Tiejun; Yu, Long-Xi; McCord, Per; Miller, David; Bhamidimarri, Suresh; Johnson, David; Monteros, Maria J; Ho, Julie; Reisen, Peter; Samac, Deborah A

    2014-01-01

    Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L.) worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  16. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L. using high-resolution melting.

    Directory of Open Access Journals (Sweden)

    Tiejun Zhang

    Full Text Available Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L. worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR and single nucleotide polymorphism (SNP markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  17. [Rapid detection of hot spot mutations of FGFR3 gene with PCR-high resolution melting assay].

    Science.gov (United States)

    Li, Shan; Wang, Han; Su, Hua; Gao, Jinsong; Zhao, Xiuli

    2017-08-10

    To identify the causative mutations in five individuals affected with dyschondroplasia and develop an efficient procedure for detecting hot spot mutations of the FGFR3 gene. Genomic DNA was extracted from peripheral blood samples with a standard phenol/chloroform method. PCR-Sanger sequencing was used to analyze the causative mutations in the five probands. PCR-high resolution melting (HRM) was developed to detect the identified mutations. A c.1138G>A mutation in exon 8 was found in 4 probands, while a c.1620C>G mutation was found in exon 11 of proband 5 whom had a mild phenotype. All patients were successfully distinguished from healthy controls with the PCR-HRM method. The results of HRM analysis were highly consistent with that of Sanger sequencing. The Gly380Arg and Asn540Lys are hot spot mutations of the FGFR3 gene among patients with ACH/HCH. PCR-HRM analysis is more efficient for detecting hot spot mutations of the FGFR3 gene.

  18. High resolution melting curve analysis, a rapid and affordable method for mutation analysis in childhood acute myeloid leukemia

    Directory of Open Access Journals (Sweden)

    Yin eLiu

    2014-09-01

    Full Text Available Background: Molecular genetic alterations with prognostic significance have been described in childhood acute myeloid leukemia (AML. The aim of this study was to establish cost-effective techniques to detect mutations of FMS-like tyrosine kinase 3 (FLT3, Nucleophosmin 1 (NPM1, and a partial tandem duplication within the mixed lineage leukemia (MLL-PTD genes in childhood AML. Procedure: Ninety-nine children with newly diagnosed AML were included in this study. We developed a fluoresent dye SYTO-82 based high resolution melting curve (HRM anaylsis to detect FLT3 internal tandem duplication (FLT3-ITD, FLT3 tyrosine kinase domain (FLT3-TKD and NPM1 mutations. MLL-PTD was screened by real-time quantitative PCR. Results: The HRM methodology correlated well with gold standard Sanger sequencing with less cost. Among the 99 patients studied, the FLT3-ITD mutation was associated with significantly worse event free survival (EFS. Patients with the NPM1 mutation had significantly better EFS and overall survival. However, HRM was not sensitive enough for minimal residual disease monitoring. Conclusions: HRM was a rapid and efficient method for screening of FLT3 and NPM1 gene mutations. It was both affordable and accurate, especially in resource underprivileged regions. Our results indicated that HRM could be a useful clinical tool for rapid and cost effective screening of the FLT3 and NPM1 mutations in AML patients.

  19. High-resolution melting analysis, a simple and effective method for reliable mutation scanning and frequency studies in the ACADVL gene

    DEFF Research Database (Denmark)

    Olsen, Rikke Katrine Jentoft; Dobrowolski, Steven F; Kjeldsen, Margrethe

    2010-01-01

    -long-chain acyl-CoA dehydrogenase deficiency (VLCADD), the second most common fatty acid oxidation disorder detected by expanded newborn screening, to demonstrate accurate and fast diagnostic evaluation of the ACADVL gene utilizing DNA extracted from the newborn screening dried blood spot and high resolution melt...

  20. High-resolution melting (HRM) analysis as a feasible method for detecting spinal muscular atrophy via dried blood spots.

    Science.gov (United States)

    Er, Tze-Kiong; Kan, Tzu-Min; Su, Yu-Fa; Liu, Ta-Chih; Chang, Jan-Gowth; Hung, Shih-Ya; Jong, Yuh-Jyh

    2012-11-12

    Spinal muscular atrophy (SMA) is a neurodegenerative disease with the leading genetic cause of infant mortality. More than 95% of patients with SMA have a homozygous disruption in the survival motor neuron1 (SMN1) gene, caused by mutation, deletion, or rearrangement. Recently, treatment in humans in the immediate postnatal period, prior to the development of weakness or very early in the course of the disease, may be effective. Therefore, our objective was to establish a feasible method for SMA screening. High-resolution melting (HRM) analysis is rapidly becoming the most important mutation-scanning methodology that allows mutation scanning and genotyping without the need for costly labeled oligonucleotides. In the current study, we aim to develop a method for identifying the substitution of single nucleotide in SMN1 exon 7 (c.840C>T) by HRM analysis. Genomic DNA was extracted from peripheral blood samples and dried blood spots obtained from 30 patients with SMA and 30 normal individuals. All results were previously confirmed by denaturing high-performance liquid chromatography (DHPLC). In order to identify the substitution of single nucleotide in SMN1 exon 7 (c.840C>T) by HRM analysis, a primer set was used in HRM analysis. At first, we failed to identify the substitution of single nucleotide in SMN1 exon 7 (c.840C>T) by HRM analysis because the homozygous CC and homozygous TT cannot be distinguished by HRM analysis. Therefore, all samples were mixed with a known SMN1/SMN2 copy number (SMN1/SMN2=0:3), which we may call driver. This strategy is used to differentiate between homozygous CC and homozygous TT. After mixing with driver, the melting profile of homozygous CC becomes heteroduplex; however, the homozygous TT remains the same in the normalized and temperature-shifted difference plots. HRM analysis can be successfully applied to screen SMA via DNA obtained from whole blood and dried blood spots. We strongly believe that HRM analysis, a high-throughput method

  1. High Resolution Melt (HRM) analysis is an efficient tool to genotype EMS mutants in complex crop genomes.

    Science.gov (United States)

    Lochlainn, Seosamh Ó; Amoah, Stephen; Graham, Neil S; Alamer, Khalid; Rios, Juan J; Kurup, Smita; Stoute, Andrew; Hammond, John P; Østergaard, Lars; King, Graham J; White, Phillip J; Broadley, Martin R

    2011-12-08

    Targeted Induced Loci Lesions IN Genomes (TILLING) is increasingly being used to generate and identify mutations in target genes of crop genomes. TILLING populations of several thousand lines have been generated in a number of crop species including Brassica rapa. Genetic analysis of mutants identified by TILLING requires an efficient, high-throughput and cost effective genotyping method to track the mutations through numerous generations. High resolution melt (HRM) analysis has been used in a number of systems to identify single nucleotide polymorphisms (SNPs) and insertion/deletions (IN/DELs) enabling the genotyping of different types of samples. HRM is ideally suited to high-throughput genotyping of multiple TILLING mutants in complex crop genomes. To date it has been used to identify mutants and genotype single mutations. The aim of this study was to determine if HRM can facilitate downstream analysis of multiple mutant lines identified by TILLING in order to characterise allelic series of EMS induced mutations in target genes across a number of generations in complex crop genomes. We demonstrate that HRM can be used to genotype allelic series of mutations in two genes, BraA.CAX1a and BraA.MET1.a in Brassica rapa. We analysed 12 mutations in BraA.CAX1.a and five in BraA.MET1.a over two generations including a back-cross to the wild-type. Using a commercially available HRM kit and the Lightscanner™ system we were able to detect mutations in heterozygous and homozygous states for both genes. Using HRM genotyping on TILLING derived mutants, it is possible to generate an allelic series of mutations within multiple target genes rapidly. Lines suitable for phenotypic analysis can be isolated approximately 8-9 months (3 generations) from receiving M3 seed of Brassica rapa from the RevGenUK TILLING service.

  2. High Resolution Melting Analysis for Detecting p53 Gene Mutations in Patients with Non-small Cell Lung Cancer

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    Zhihong CHEN

    2011-10-01

    Full Text Available Background and objective It has been proven that p53 gene was related to many human cancers. The mutations in p53 gene play an important role in carcinogensis and mostly happened in exon 5-8. The aim of this study is to establish a high resolution melting (HRM assay to detect p53 mutations from patients with non-small cell lung cancer (NSCLC, to investigate the characteristics of p53 gene mutations, and to analyze the relationship between p53 mutations and evolution regularity of pathogenesis. Methods p53 mutations in exon 5-8 were detected by HRM assay on DNA insolated from 264 NSCLC samples derived from tumor tissues and 54 control samples from pericancerous pulmonary tissues. The mutation samples by the HRM assay were confirmed by sequencing technique. Samples which were positive by HRM but wild type by sequencing were further confirmed by sub-clone and sequencing. Results No mutation was found in 54 pericancerous pulmonary samples by HRM assay. 104 of the 264 tumor tissues demonstrated mutation curves by HRM assay, 102 samples were confirmed by sequencing, including 95 point mutations and 7 frame shift mutations by insertion or deletion. The mutation rate of p53 gene was 39.4%. The mutation rate from exon 5-8 were 11.7%, 8%, 12.5% and 10.6%, respectively and there was no statistically significant difference between them (P=0.35. p53 mutations were significantly more frequent in males than that in females, but not related to the other clinicopathologic characteristics. Conclusion The results indicate that HRM is a sensitive in-tube methodology to detect for mutations in clinical samples. The results suggest that the arising p53 mutations in NSCLC may be due to spontaneous error in DNA synthesis and repair.

  3. Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2

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    Sinilnikova Olga

    2011-05-01

    Full Text Available Abstract Background The gene CHEK2 encodes a checkpoint kinase playing a key role in the DNA damage pathway. Though CHEK2 has been identified as an intermediate breast cancer susceptibility gene, only a small proportion of high-risk families have been explained by genetic variants located in its coding region. Alteration in gene expression regulation provides a potential mechanism for generating disease susceptibility. The detection of differential allelic expression (DAE represents a sensitive assay to direct the search for a functional sequence variant within the transcriptional regulatory elements of a candidate gene. We aimed to assess whether CHEK2 was subject to DAE in lymphoblastoid cell lines (LCLs from high-risk breast cancer patients for whom no mutation in BRCA1 or BRCA2 had been identified. Methods We implemented an assay based on high-resolution melting (HRM curve analysis and developed an analysis tool for DAE assessment. Results We observed allelic expression imbalance in 4 of the 41 LCLs examined. All four were carriers of the truncating mutation 1100delC. We confirmed previous findings that this mutation induces non-sense mediated mRNA decay. In our series, we ruled out the possibility of a functional sequence variant located in the promoter region or in a regulatory element of CHEK2 that would lead to DAE in the transcriptional regulatory milieu of freely proliferating LCLs. Conclusions Our results support that HRM is a sensitive and accurate method for DAE assessment. This approach would be of great interest for high-throughput mutation screening projects aiming to identify genes carrying functional regulatory polymorphisms.

  4. High Resolution Melt (HRM analysis is an efficient tool to genotype EMS mutants in complex crop genomes

    Directory of Open Access Journals (Sweden)

    Lochlainn Seosamh Ó

    2011-12-01

    Full Text Available Abstract Background Targeted Induced Loci Lesions IN Genomes (TILLING is increasingly being used to generate and identify mutations in target genes of crop genomes. TILLING populations of several thousand lines have been generated in a number of crop species including Brassica rapa. Genetic analysis of mutants identified by TILLING requires an efficient, high-throughput and cost effective genotyping method to track the mutations through numerous generations. High resolution melt (HRM analysis has been used in a number of systems to identify single nucleotide polymorphisms (SNPs and insertion/deletions (IN/DELs enabling the genotyping of different types of samples. HRM is ideally suited to high-throughput genotyping of multiple TILLING mutants in complex crop genomes. To date it has been used to identify mutants and genotype single mutations. The aim of this study was to determine if HRM can facilitate downstream analysis of multiple mutant lines identified by TILLING in order to characterise allelic series of EMS induced mutations in target genes across a number of generations in complex crop genomes. Results We demonstrate that HRM can be used to genotype allelic series of mutations in two genes, BraA.CAX1a and BraA.MET1.a in Brassica rapa. We analysed 12 mutations in BraA.CAX1.a and five in BraA.MET1.a over two generations including a back-cross to the wild-type. Using a commercially available HRM kit and the Lightscanner™ system we were able to detect mutations in heterozygous and homozygous states for both genes. Conclusions Using HRM genotyping on TILLING derived mutants, it is possible to generate an allelic series of mutations within multiple target genes rapidly. Lines suitable for phenotypic analysis can be isolated approximately 8-9 months (3 generations from receiving M3 seed of Brassica rapa from the RevGenUK TILLING service.

  5. High resolution melting analysis for the detection of EMS induced mutations in wheat SbeIIa genes

    Directory of Open Access Journals (Sweden)

    Botticella Ermelinda

    2011-11-01

    Full Text Available Abstract Background Manipulation of the amylose-amylopectin ratio in cereal starch has been identified as a major target for the production of starches with novel functional properties. In wheat, silencing of starch branching enzyme genes by a transgenic approach reportedly caused an increase of amylose content up to 70% of total starch, exhibiting novel and interesting nutritional characteristics. In this work, the functionality of starch branching enzyme IIa (SBEIIa has been targeted in bread wheat by TILLING. An EMS-mutagenised wheat population has been screened using High Resolution Melting of PCR products to identify functional SNPs in the three homoeologous genes encoding the target enzyme in the hexaploid genome. Results This analysis resulted in the identification of 56, 14 and 53 new allelic variants respectively for SBEIIa-A, SBEIIa-B and SBEIIa-D. The effects of the mutations on protein structure and functionality were evaluated by a bioinformatic approach. Two putative null alleles containing non-sense or splice site mutations were identified for each of the three homoeologous SBEIIa genes; qRT-PCR analysis showed a significant decrease of their gene expression and resulted in increased amylose content. Pyramiding of different single null homoeologous allowed to isolate double null mutants showing an increase of amylose content up to 21% compared to the control. Conclusion TILLING has successfully been used to generate novel alleles for SBEIIa genes known to control amylose content in wheat. Single and double null SBEIIa genotypes have been found to show a significant increase in amylose content.

  6. High-resolution Melting Analysis for Gene Scanning of Adenomatous Polyposis Coli (APC) Gene With Oral Squamous Cell Carcinoma Samples.

    Science.gov (United States)

    Chang, Ya-Sian; Lin, Chien-Yu; Yang, Shu-Fen; Ho, Cheng Mao; Chang, Jan-Gowth

    2016-02-01

    There have been many different mutations reported for the large adenomatous polyposis coli (APC) tumor suppressor gene. APC mutations result in inactivation of APC tumor suppressor action, allowing the progression of tumorigenesis. The present study utilized a highly efficient method to identify APC mutations and investigated the association between the APC genetic variants Y486Y, A545A, T1493T, and D1822V and susceptibility to oral squamous cell carcinoma (OSCC). High-resolution melting (HRM) analysis was used to characterize APC mutations. Genomic DNA was extracted from 83 patient specimens of OSCC and 50 blood samples from healthy control subjects. The 14 exons and mutation cluster region of exon 15 were screened by HRM analysis. All mutations were confirmed by direct DNA sequencing. Three mutations and 4 single nucleotide polymorphisms (SNPs) were found in this study. The mutations were c.573T>C (Y191Y) in exon 5, c.1005A>G (L335L) in exon 9, and c.1488A>T (T496T) in exon 11. Two SNPs, c.4479G>A (T1493T) and c.5465A>T (D1822V), were located in exon 15, whereas c.1458T>C (Y486Y) and c.1635G>A (A545A) were located in exon 11 and 13, respectively. There was no observed association between OSCC risk and genotype for any of the 4 APC SNPs. The mutation of APC is rare in Taiwanese patients with OSCC. HRM analysis is a reliable, accurate, and fast screening method for APC mutations.

  7. SNP discovery and High Resolution Melting Analysis from massive transcriptome sequencing in the California red abalone Haliotis rufescens.

    Science.gov (United States)

    Valenzuela-Muñoz, Valentina; Araya-Garay, José Miguel; Gallardo-Escárate, Cristian

    2013-06-01

    The California red abalone, Haliotis rufescens that belongs to the Haliotidae family, is the largest species of abalone in the world that has sustained the major fishery and aquaculture production in the USA and Mexico. This native mollusk has not been evaluated or assigned a conservation category even though in the last few decades it was heavily exploited until it disappeared in some areas along the California coast. In Chile, the red abalone was introduced in the 1970s from California wild abalone stocks for the purposes of aquaculture. Considering the number of years that the red abalone has been cultivated in Chile crucial genetic information is scarce and critical issues remain unresolved. This study reports and validates novel single nucleotide polymorphisms (SNP) markers for the red abalone H. rufescens using cDNA pyrosequencing. A total of 622 high quality SNPs were identified in 146 sequences with an estimated frequency of 1 SNP each 1000bp. Forty-five SNPs markers with functional information for gene ontology were selected. Of these, 8 were polymorphic among the individuals screened: Heat shock protein 70 (HSP70), vitellogenin (VTG), lysin, alginate lyase enzyme (AL), Glucose-regulated protein 94 (GRP94), fructose-bisphosphate aldolase (FBA), sulfatase 1A precursor (S1AP) and ornithine decarboxylase antizyme (ODC). Two additional sequences were also identified with polymorphisms but no similarities with known proteins were achieved. To validate the putative SNP markers, High Resolution Melting Analysis (HRMA) was conducted in a wild and hatchery-bred population. Additionally, SNP cross-amplifications were tested in two further native abalone species, Haliotis fulgens and Haliotis corrugata. This study provides novel candidate genes that could be used to evaluate loss of genetic diversity due to hatchery selection or inbreeding effects. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Analysis of HIV using a high resolution melting (HRM) diversity assay: automation of HRM data analysis enhances the utility of the assay for analysis of HIV incidence.

    Science.gov (United States)

    Cousins, Matthew M; Swan, David; Magaret, Craig A; Hoover, Donald R; Eshleman, Susan H

    2012-01-01

    HIV diversity may be a useful biomarker for discriminating between recent and non-recent HIV infection. The high resolution melting (HRM) diversity assay was developed to quantify HIV diversity in viral populations without sequencing. In this assay, HIV diversity is expressed as a single numeric HRM score that represents the width of a melting peak. HRM scores are highly associated with diversity measures obtained with next generation sequencing. In this report, a software package, the HRM Diversity Assay Analysis Tool (DivMelt), was developed to automate calculation of HRM scores from melting curve data. DivMelt uses computational algorithms to calculate HRM scores by identifying the start (T1) and end (T2) melting temperatures for a DNA sample and subtracting them (T2 - T1 =  HRM score). DivMelt contains many user-supplied analysis parameters to allow analyses to be tailored to different contexts. DivMelt analysis options were optimized to discriminate between recent and non-recent HIV infection and to maximize HRM score reproducibility. HRM scores calculated using DivMelt were compared to HRM scores obtained using a manual method that is based on visual inspection of DNA melting curves. HRM scores generated with DivMelt agreed with manually generated HRM scores obtained from the same DNA melting data. Optimal parameters for discriminating between recent and non-recent HIV infection were identified. DivMelt provided greater discrimination between recent and non-recent HIV infection than the manual method. DivMelt provides a rapid, accurate method of determining HRM scores from melting curve data, facilitating use of the HRM diversity assay for large-scale studies.

  9. High-resolution melt PCR analysis for rapid identification of Chlamydia abortus live vaccine strain 1B among C. abortus strains and field isolates.

    Science.gov (United States)

    Vorimore, Fabien; Cavanna, Noémie; Vicari, Nadia; Magnino, Simone; Willems, Hermann; Rodolakis, Annie; Siarkou, Victoria I; Laroucau, Karine

    2012-09-01

    We describe a novel high-resolution melt assay that clearly differentiates Chlamydia abortus live vaccine strain 1B from field C. abortus strains and field wild-type isolates based on previously described single nucleotide polymorphisms. This modern genotyping technique is inexpensive, easy to use, and less time-consuming than PCR-RFLP. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Rapid analysis of heterogeneously methylated DNA using digital methylation-sensitive high resolution melting: application to the CDKN2B (p15) gene

    DEFF Research Database (Denmark)

    Candiloro, Ida Lm; Mikeska, Thomas; Hokland, Peter

    2008-01-01

    ABSTRACT: BACKGROUND: Methylation-sensitive high resolution melting (MS-HRM) methodology is able to recognise heterogeneously methylated sequences by their characteristic melting profiles. To further analyse heterogeneously methylated sequences, we adopted a digital approach to MS-HRM (dMS-HRM) t......ABSTRACT: BACKGROUND: Methylation-sensitive high resolution melting (MS-HRM) methodology is able to recognise heterogeneously methylated sequences by their characteristic melting profiles. To further analyse heterogeneously methylated sequences, we adopted a digital approach to MS-HRM (d......MS-HRM) that involves the amplification of single templates after limiting dilution to quantify and to determine the degree of methylation. We used this approach to study methylation of the CDKN2B (p15) cell cycle progression inhibitor gene which is inactivated by DNA methylation in haematological malignancies...... the methylated alleles and assess the degree of methylation. Direct sequencing of selected dMS-HRM products was used to determine the exact DNA methylation pattern and confirmed the degree of methylation estimated by dMS-HRM. CONCLUSION: dMS-HRM is a powerful technique for the analysis of methylation in CDKN2B...

  11. Using high-resolution tritium profiles to quantify the effects of melt on two Spitsbergen ice cores

    NARCIS (Netherlands)

    Wel, L.G. van der; Streurman, H.J.; Isaksson, E.; Helsen, M.M.; Wal, R.S.W. van de; Martma, T.; Pohjola, V.A.; Moore, J.C.; Meijer, H.A.J.

    2011-01-01

    Ice cores from small ice caps provide valuable climatic information, additional to that of Greenland and Antarctica. However, their integrity is usually compromised by summer meltwater percolation. To determine to what extent this can affect such ice cores, we performed high-resolution tritium

  12. Hybrid analysis (barcode-high resolution melting) for authentication of Thai herbal products, Andrographis paniculata (Burm.f.) Wall.ex Nees.

    Science.gov (United States)

    Osathanunkul, Maslin; Suwannapoom, Chatmongkon; Khamyong, Nuttaluck; Pintakum, Danupol; Lamphun, Santisuk Na; Triwitayakorn, Kanokporn; Osathanunkul, Kitisak; Madesis, Panagiotis

    2016-01-01

    Andrographis paniculata Nees is a medicinal plant with multiple pharmacological properties. It has been used over many centuries as a household remedy. A. paniculata products sold on the markets are in processed forms so it is difficult to authenticate. Therefore buying the herbal products poses a high-risk of acquiring counterfeited, substituted and/or adulterated products. Due to these issues, a reliable method to authenticate products is needed. High resolution melting analysis coupled with DNA barcoding (Bar-HRM) was applied to detect adulteration in commercial herbal products. The rbcL barcode was selected to use in primers design for HRM analysis to produce standard melting profile of A. paniculata species. DNA of the tested commercial products was isolated and their melting profiles were then generated and compared with the standard A. paniculata. The melting profiles of the rbcL amplicons of the three closely related herbal species (A. paniculata, Acanthus ebracteatus and Rhinacanthus nasutus) are clearly separated so that they can be distinguished by the developed method. The method was then used to authenticate commercial herbal products. HRM curves of all 10 samples tested are similar to A. paniculata which indicated that all tested products were contained the correct species as labeled. The method described in this study has been proved to be useful in aiding identification and/or authenticating A. paniculata. This Bar-HRM analysis has allowed us easily to determine the A. paniculata species in herbal products on the markets even they are in processed forms. We propose the use of DNA barcoding combined with High Resolution Melting analysis for authenticating of Andrographis paniculata products.The developed method can be used regardless of the type of the DNA template (fresh or dried tissue, leaf, and stem).rbcL region was chosen for the analysis and work well with our samplesWe can easily determine the A. paniculata species in herbal products tested

  13. Detection of plant oil DNA using high resolution melting (HRM) post PCR analysis: a tool for disclosure of olive oil adulteration.

    Science.gov (United States)

    Vietina, Michelangelo; Agrimonti, Caterina; Marmiroli, Nelson

    2013-12-15

    Extra virgin olive oil is frequently subjected to adulterations with addition of oils obtained from plants other than olive. DNA analysis is a fast and economic tool to identify plant components in oils. Extraction and amplification of DNA by PCR was tested in olives, in milled seeds and in oils, to investigate its use in olive oil traceability. DNA was extracted from different oils made of hazelnut, maize, sunflower, peanut, sesame, soybean, rice and pumpkin. Comparing the DNA melting profiles in reference plant materials and in the oils, it was possible to identify any plant components in oils and mixtures of oils. Real-Time PCR (RT-PCR) platform has been added of the new methodology of high resolution melting (HRM), both were used to analyse olive oils mixed with different percentage of other oils. Results showed HRM a cost effective method for efficient detection of adulterations in olive oils. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.

    Science.gov (United States)

    Dehghani, Mansoureh; Mohammadi, Mohammad Ali; Rostami, Sima; Shamsaddini, Saeedeh; Mirbadie, Seyed Reza; Harandi, Majid Fasihi

    2016-07-01

    Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.

  15. High Resolution Melting (HRM) analysis for mutation screening of RGSL1,RGS16 and RGS8 in breast cancer

    DEFF Research Database (Denmark)

    Wiechec, Emilia; Wiuf, Carsten Henrik; Overgaard, Jens

    2011-01-01

    coding exons of RGSL1, RGS16, and RGS8 in tumors from 200 breast cancer patients. All sequence variants detected by HRM resulted in abnormal shape of the melting curves. The identified mutations and known single nucleotide polymorphisms (SNP) were subsequently confirmed by sequencing, and distribution...... cancer patients. In addition, a total of seven known SNPs were identified in this study. Genotype distributions were not significantly different between breast cancer patients and controls. CONCLUSIONS AND IMPACT: Identification of novel mutations within RGSL1 provides a new insight...... into the pathophysiology of breast cancer. Moreover, the HRM analysis represents a reliable and highly sensitive method for mutation scanning of multiple exons....

  16. Product differentiation by analysis of DNA melting curves during the polymerase chain reaction.

    Science.gov (United States)

    Ririe, K M; Rasmussen, R P; Wittwer, C T

    1997-02-15

    A microvolume fluorometer integrated with a thermal cycler was used to acquire DNA melting curves during polymerase chain reaction by fluorescence monitoring of the double-stranded DNA specific dye SYBR Green I. Plotting fluorescence as a function of temperature as the thermal cycler heats through the dissociation temperature of the product gives a DNA melting curve. The shape and position of this DNA melting curve are functions of the GC/AT ratio, length, and sequence and can be used to differentiate amplification products separated by less than 2 degrees C in melting temperature. Desired products can be distinguished from undesirable products, in many cases eliminating the need for gel electrophoresis. Analysis of melting curves can extend the dynamic range of initial template quantification when amplification is monitored with double-stranded DNA specific dyes. Complete amplification and analysis of products can be performed in less than 15 min.

  17. Assessment of genetic mutations in the XRCC2 coding region by high resolution melting curve analysis and the risk of differentiated thyroid carcinoma in Iran

    Directory of Open Access Journals (Sweden)

    Shima Fayaz

    2012-01-01

    Full Text Available Homologous recombination (HR is the major pathway for repairing double strand breaks (DSBs in eukaryotes and XRCC2 is an essential component of the HR repair machinery. To evaluate the potential role of mutations in gene repair by HR in individuals susceptible to differentiated thyroid carcinoma (DTC we used high resolution melting (HRM analysis, a recently introduced method for detecting mutations, to examine the entire XRCC2 coding region in an Iranian population. HRM analysis was used to screen for mutations in three XRCC2 coding regions in 50 patients and 50 controls. There was no variation in the HRM curves obtained from the analysis of exons 1 and 2 in the case and control groups. In exon 3, an Arg188His polymorphism (rs3218536 was detected as a new melting curve group (OR: 1.46; 95%CI: 0.432-4.969; p = 0.38 compared with the normal melting curve. We also found a new Ser150Arg polymorphism in exon 3 of the control group. These findings suggest that genetic variations in the XRCC2 coding region have no potential effects on susceptibility to DTC. However, further studies with larger populations are required to confirm this conclusion.

  18. Rapid screening of ASXL1, IDH1, IDH2, and c-CBL mutations in de novo acute myeloid leukemia by high-resolution melting.

    Science.gov (United States)

    Ibáñez, Mariam; Such, Esperanza; Cervera, José; Luna, Irene; Gómez-Seguí, Inés; López-Pavía, María; Dolz, Sandra; Barragán, Eva; Fuster, Oscar; Llop, Marta; Rodríguez-Veiga, Rebeca; Avaria, Amparo; Oltra, Silvestre; Senent, M Leonor; Moscardó, Federico; Montesinos, Pau; Martínez-Cuadrón, David; Martín, Guillermo; Sanz, Miguel A

    2012-11-01

    Recently, many novel molecular abnormalities were found to be distinctly associated with acute myeloid leukemia (AML). However, their clinical relevance and prognostic implications are not well established. We developed a new combination of high-resolution melting assays on a LightCycler 480 and direct sequencing to detect somatic mutations of ASXL1 (exon 12), IDH1 (exon 4), IDH2 (exon 4), and c-CBL (exons 8 and 9) genes to know their incidence and prognostic effect in a cohort of 175 patients with de novo AML: 16 patients (9%) carried ASXL1 mutations, 16 patients had IDH variations (3% with IDH1(R132) and 6% with IDH2(R140)), and none had c-CBL mutations. Patients with ASXL1 mutations did not harbor IDH1, [corrected] or CEBPA mutations, and a combination of ASXL1 and IDH2 mutations was found only in one patient. In addition, we did not find IDH1 and FLT3 or CEBPA mutations concurrently or IDH2 with CEBPA. IDH1 and IDH2 mutations were mutually exclusive. Alternatively, NPM1 mutations were concurrently found with ASXL1, IDH1, or IDH2 with a variable incidence. Mutations were not significantly correlated with any of the clinical and biological features studied. High-resolution melting is a reliable, rapid, and efficient screening technique for mutation detection in AML. The incidence for the studied genes was in the range of those previously reported. We were unable to find an effect on the outcome. Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  19. Authenticity analyses of Rhizoma Paridis using barcoding coupled with high resolution melting (Bar-HRM) analysis to control its quality for medicinal plant product.

    Science.gov (United States)

    Duan, Bao-Zhong; Wang, Ya-Ping; Fang, Hai-Lan; Xiong, Chao; Li, Xi-Wen; Wang, Ping; Chen, Shi-Lin

    2018-01-01

    Rhizoma Paridis (Chonglou) is a commonly used and precious traditional Chinese medicine. Paris polyphylla Smith var. yunnanensis (Franch.) Hand. -Mazz. and Paris polyphylla Smith var . chinensis (Franch.) Hara are the two main sources of Chonglou under the monograph of Rhizoma Paridis in Chinese Pharmacopoeia. In the local marketplace, however, this medicine is prone to be accidentally contaminated, deliberately substituted or admixed with other species that are similar to Rhizoma Paridis in shape and color. Consequently, these adulterations might compromise quality control and result in considerable health concerns for consumers. This study aims to develop a rapid and sensitive method for accurate identification of Rhizoma Paridis and its common adulterants. DNA barcoding coupled with high resolution melting analysis was applied in this research to distinguish Rhizoma Paridis from its adulteration. The internal transcribed spacer 2 (ITS2) barcode was selected for HRM analysis to produce standard melting profile of the selected species. DNA of the tested herbal medicines was isolated and their melting profiles were generated and compared with the standard melting profile of P. polyphylla var. chinensis . The results indicate that the ITS2 molecular regions coupled with HRM analysis can effectively differentiate nine herbal species, including two authentic origins of Chonglou and their seven common adulterants. Ten herbal medicines labeled "Chonglou" obtained from a local market were collected and identified with our methods, and their sequence information was analyzed to validate the accuracy of HRM analysis. DNA barcoding coupled with HRM analysis is a accurate, reliable, rapid, cost-effective and robust tool, which could contribute to the quality control of Rhizoma Paridis in the supply chain of the natural health product industry (NHP).

  20. High-resolution melting analysis for detection of a single-nucleotide polymorphism and the genotype of the myostatin gene in warmblood horses.

    Science.gov (United States)

    Serpa, Priscila B S; Garbade, Petra; Natalini, Cláudio C; Pires, Ananda R; Tisotti, Tainor M

    2017-01-01

    OBJECTIVE To develop a high-resolution melting (HRM) assay to detect the g.66493737C>T polymorphism in the myostatin gene (MSTN) and determine the frequency of 3 previously defined g.66493737 genotypes (T/T, T/C, and C/C) in warmblood horses. SAMPLES Blood samples from 23 horses. PROCEDURES From each blood sample, DNA was extracted and analyzed by standard PCR methods and an HRM assay to determine the MSTN genotype. Three protocols (standard protocol, protocol in which a high-salt solution was added to the reaction mixture before the first melting cycle, and protocol in which an unlabeled probe was added to the reaction mixture before analysis) for the HRM assay were designed and compared. Genotype results determined by the HRM protocol that generated the most consistent melting curves were compared with those determined by sequencing. RESULTS The HRM protocol in which an unlabeled probe was added to the reaction mixture generated the most consistent melting curves. The genotypes of the g.66493737C>T polymorphism were determined for 22 horses (16 by HRM analysis and 20 by sequencing); 14, 7, and 1 had the T/T, T/C, and C/C genotypes, respectively. The genotype determined by HRM analysis agreed with that determined by sequencing for 14 of 16 horses. The frequency of alleles T and C was 79.5% and 20.5%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that HRM analysis may be a faster and more economical alternative than PCR methods for genotyping. Genotyping results might be useful as predictors of athletic performance for horses.

  1. A comparison of PCR assays for beak and feather disease virus and high resolution melt (HRM) curve analysis of replicase associated protein and capsid genes.

    Science.gov (United States)

    Das, Shubhagata; Sarker, Subir; Ghorashi, Seyed Ali; Forwood, Jade K; Raidal, Shane R

    2016-11-01

    Beak and feather disease virus (BFDV) threatens a wide range of endangered psittacine birds worldwide. In this study, we assessed a novel PCR assay and genetic screening method using high-resolution melt (HRM) curve analysis for BFDV targeting the capsid (Cap) gene (HRM-Cap) alongside conventional PCR detection as well as a PCR method that targets a much smaller fragment of the virus genome in the replicase initiator protein (Rep) gene (HRM-Rep). Limits of detection, sensitivity, specificity and discriminatory power for differentiating BFDV sequences were compared. HRM-Cap had a high positive predictive value and could readily differentiate between a reference genotype and 17 other diverse BFDV genomes with more discriminatory power (genotype confidence percentage) than HRM-Rep. Melt curve profiles generated by HRM-Cap correlated with unique DNA sequence profiles for each individual test genome. The limit of detection of HRM-Cap was lower (2×10 -5 ng/reaction or 48 viral copies) than that for both HRM-Rep and conventional BFDV PCR which had similar sensitivity (2×10 -6 ng or 13 viral copies/reaction). However, when used in a diagnostic setting with 348 clinical samples there was strong agreement between HRM-Cap and conventional PCR (kappa=0.87, PHRM-Cap demonstrated higher specificity (99.9%) than HRM-Rep (80.3%). Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Development of cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers from the chloroplast genome of Glycyrrhiza species.

    Science.gov (United States)

    Jo, Ick-Hyun; Sung, Jwakyung; Hong, Chi-Eun; Raveendar, Sebastin; Bang, Kyong-Hwan; Chung, Jong-Wook

    2018-05-01

    Licorice ( Glycyrrhiza glabra ) is an important medicinal crop often used as health foods or medicine worldwide. The molecular genetics of licorice is under scarce owing to lack of molecular markers. Here, we have developed cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers based on single nucleotide polymorphisms (SNP) by comparing the chloroplast genomes of two Glycyrrhiza species ( G. glabra and G. lepidota ). The CAPS and HRM markers were tested for diversity analysis with 24 Glycyrrhiza accessions. The restriction profiles generated with CAPS markers classified the accessions (2-4 genotypes) and melting curves (2-3) were obtained from the HRM markers. The number of alleles and major allele frequency were 2-6 and 0.31-0.92, respectively. The genetic distance and polymorphism information content values were 0.16-0.76 and 0.15-0.72, respectively. The phylogenetic relationships among the 24 accessions were estimated using a dendrogram, which classified them into four clades. Except clade III, the remaining three clades included the same species, confirming interspecies genetic correlation. These 18 CAPS and HRM markers might be helpful for genetic diversity assessment and rapid identification of licorice species.

  3. High-resolution melting of 12S rRNA and cytochrome b DNA sequences for discrimination of species within distinct European animal families.

    Directory of Open Access Journals (Sweden)

    Jana Naue

    Full Text Available The cheap and easy identification of species is necessary within multiple fields of molecular biology. The use of high-resolution melting (HRM of DNA provides a fast closed-tube method for analysis of the sequence composition of the mitochondrial genes 12S rRNA and cytochrome b. We investigated the potential use of HRM for species identification within eleven different animal groups commonly found in Europe by animal-group-specific DNA amplification followed by DNA melting. Influence factors as DNA amount, additional single base alterations, and the existence of mixed samples were taken into consideration. Visual inspection combined with mathematical evaluation of the curve shapes did resolve nearly all species within an animal group. The assay can therefore not only be used for identification of animal groups and mixture analysis but also for species identification within the respective groups. The use of a universal 12S rRNA system additionally revealed a possible approach for species discrimination, mostly by exclusion. The use of the HRM assay showed to be a reliable, fast, and cheap method for species discrimination within a broad range of different animal species and can be used in a flexible "modular" manner depending on the question to be solved.

  4. Candidate DNA Barcode Tags Combined With High Resolution Melting (Bar-HRM Curve Analysis for Authentication of Senna alexandrina Mill. With Validation in Crude Drugs

    Directory of Open Access Journals (Sweden)

    Priyanka Mishra

    2018-03-01

    Full Text Available Senna alexandrina (Fabaceae is a globally recognized medicinal plant for its laxative properties as well as the only source of sennosides, and is highly exported bulk herb from India. Its major procurement is exclusively from limited cultivation, which leads to risks of deliberate or unintended adulteration. The market raw materials are in powdered or finished product form, which lead to difficulties in authentication. Here, DNA barcode tags based on chloroplast genes (rbcL and matK and intergenic spacers (psbA-trnH and ITS were developed for S. alexandrina along with the allied species. The ability and performance of the ITS1 region to discriminate among the Senna species resulted in the present proposal of the ITS1 tags as successful barcode. Further, these tags were coupled with high-resolution melting (HRM curve analysis in a real-time PCR genotyping method to derive Bar-HRM (Barcoding-HRM assays. Suitable HRM primer sets were designed through SNP detection and mutation scanning in genomic signatures of Senna species. The melting profiles of S. alexandrina and S. italica subsp. micrantha were almost identical and the remaining five species were clearly separated so that they can be differentiated by HRM method. The sensitivity of the method was utilized to authenticate market samples [Herbal Sample Assays (HSAs]. HSA01 (S. alexandrina crude drug sample from Bangalore and HSA06 (S. alexandrina crude drug sample from Tuticorin, Tamil Nadu, India were found to be highly contaminated with S. italica subsp. micrantha. Species admixture samples mixed in varying percentage was identified sensitively with detection of contamination as low as 1%. The melting profiles of PCR amplicons are clearly distinct, which enables the authentic differentiation of species by the HRM method. This study reveals that DNA barcoding coupled with HRM is an efficient molecular tool to authenticate Senna herbal products in the market for quality control in the drug

  5. Candidate DNA Barcode Tags Combined With High Resolution Melting (Bar-HRM) Curve Analysis for Authentication of Senna alexandrina Mill. With Validation in Crude Drugs.

    Science.gov (United States)

    Mishra, Priyanka; Shukla, Ashutosh K; Sundaresan, Velusamy

    2018-01-01

    Senna alexandrina (Fabaceae) is a globally recognized medicinal plant for its laxative properties as well as the only source of sennosides, and is highly exported bulk herb from India. Its major procurement is exclusively from limited cultivation, which leads to risks of deliberate or unintended adulteration. The market raw materials are in powdered or finished product form, which lead to difficulties in authentication. Here, DNA barcode tags based on chloroplast genes ( rbcL and matK ) and intergenic spacers ( psbA-trnH and ITS ) were developed for S. alexandrina along with the allied species. The ability and performance of the ITS1 region to discriminate among the Senna species resulted in the present proposal of the ITS1 tags as successful barcode. Further, these tags were coupled with high-resolution melting (HRM) curve analysis in a real-time PCR genotyping method to derive Bar-HRM (Barcoding-HRM) assays. Suitable HRM primer sets were designed through SNP detection and mutation scanning in genomic signatures of Senna species. The melting profiles of S. alexandrina and S . italica subsp. micrantha were almost identical and the remaining five species were clearly separated so that they can be differentiated by HRM method. The sensitivity of the method was utilized to authenticate market samples [Herbal Sample Assays (HSAs)]. HSA01 ( S. alexandrina crude drug sample from Bangalore) and HSA06 ( S. alexandrina crude drug sample from Tuticorin, Tamil Nadu, India) were found to be highly contaminated with S . italica subsp. micrantha . Species admixture samples mixed in varying percentage was identified sensitively with detection of contamination as low as 1%. The melting profiles of PCR amplicons are clearly distinct, which enables the authentic differentiation of species by the HRM method. This study reveals that DNA barcoding coupled with HRM is an efficient molecular tool to authenticate Senna herbal products in the market for quality control in the drug supply

  6. Identification and Differentiation of Monilinia Species Causing Brown Rot of Pome and Stone Fruit using High-Resolution Melting (HRM) Analysis.

    Science.gov (United States)

    Papavasileiou, Antonios; Madesis, Panagiotis B; Karaoglanidis, George S

    2016-09-01

    Brown rot is a devastating disease of stone fruit caused by Monilinia spp. Among these species, Monilinia fructicola is a quarantine pathogen in Europe but has recently been detected in several European countries. Identification of brown rot agents relies on morphological differences or use of molecular methods requiring fungal isolation. The current study was initiated to develop and validate a high-resolution melting (HRM) method for the identification of the Monilinia spp. and for the detection of M. fructicola among other brown rot pathogens. Based on the sequence of the cytb intron from M. laxa, M. fructicola, M. fructigena, M. mumecola, M. linhartiana, and M. yunnanensis isolates originating from several countries, a pair of universal primers for species identification and a pair of primers specific to M. fructicola were designed. The specificity of the primers was verified to ensure against cross-reaction with other fungal species. The melting curve analysis using the universal primers generated six different HRM curve profiles, each one specific for each species. Τhe HRM analysis primers specific to M. fructicola amplified a 120-bp region with a distinct melt profile corresponding to the presence of M. fructicola, regardless of the presence of other species. HRM analysis can be a useful tool for rapid identification and differentiation of the six Monilinia spp. using a single primer pair. This novel assay has the potential for simultaneous identification and differentiation of the closely related Monilinia spp. as well as for the differentiation of M. fructicola from other common pathogens or saprophytes that may occur on the diseased fruit.

  7. Unraveling Host-Vector-Arbovirus Interactions by Two-Gene High Resolution Melting Mosquito Bloodmeal Analysis in a Kenyan Wildlife-Livestock Interface.

    Directory of Open Access Journals (Sweden)

    David Omondi

    Full Text Available The blood-feeding patterns of mosquitoes are directly linked to the spread of pathogens that they transmit. Efficient identification of arthropod vector bloodmeal hosts can identify the diversity of vertebrate species potentially involved in disease transmission cycles. While molecular bloodmeal analyses rely on sequencing of cytochrome b (cyt b or cytochrome oxidase 1 gene PCR products, recently developed bloodmeal host identification based on high resolution melting (HRM analyses of cyt b PCR products is more cost-effective. To resolve the diverse vertebrate hosts that mosquitoes may potentially feed on in sub-Saharan Africa, we utilized HRM profiles of both cyt b and 16S ribosomal RNA genes. Among 445 blood-fed Aedeomyia, Aedes, Anopheles, Culex, Mansonia, and Mimomyia mosquitoes from Kenya's Lake Victoria and Lake Baringo regions where many mosquito-transmitted pathogens are endemic, we identified 33 bloodmeal hosts including humans, eight domestic animal species, six peridomestic animal species and 18 wildlife species. This resolution of vertebrate host species was only possible by comparing profiles of both cyt b and 16S markers, as melting profiles of some pairs of species were similar for either marker but not both. We identified mixed bloodmeals in a Culex pipiens from Mbita that had fed on a goat and a human and in two Mansonia africana mosquitoes from Baringo that each had fed on a rodent (Arvicanthis niloticus in addition to a human or baboon. We further detected Sindbis and Bunyamwera viruses in blood-fed mosquito homogenates by Vero cell culture and RT-PCR in Culex, Aedeomyia, Anopheles and Mansonia mosquitoes from Baringo that had fed on humans and livestock. The observed mosquito feeding on both arbovirus amplifying hosts (including sheep and goats and possible arbovirus reservoirs (birds, porcupine, baboons, rodents informs arbovirus disease epidemiology and vector control strategies.

  8. Use of a high resolution melting (HRM assay to compare gag, pol, and env diversity in adults with different stages of HIV infection.

    Directory of Open Access Journals (Sweden)

    Matthew M Cousins

    Full Text Available Cross-sectional assessment of HIV incidence relies on laboratory methods to discriminate between recent and non-recent HIV infection. Because HIV diversifies over time in infected individuals, HIV diversity may serve as a biomarker for assessing HIV incidence. We used a high resolution melting (HRM diversity assay to compare HIV diversity in adults with different stages of HIV infection. This assay provides a single numeric HRM score that reflects the level of genetic diversity of HIV in a sample from an infected individual.HIV diversity was measured in 203 adults: 20 with acute HIV infection (RNA positive, antibody negative, 116 with recent HIV infection (tested a median of 189 days after a previous negative HIV test, range 14-540 days, and 67 with non-recent HIV infection (HIV infected >2 years. HRM scores were generated for two regions in gag, one region in pol, and three regions in env.Median HRM scores were higher in non-recent infection than in recent infection for all six regions tested. In multivariate models, higher HRM scores in three of the six regions were independently associated with non-recent HIV infection.The HRM diversity assay provides a simple, scalable method for measuring HIV diversity. HRM scores, which reflect the genetic diversity in a viral population, may be useful biomarkers for evaluation of HIV incidence, particularly if multiple regions of the HIV genome are examined.

  9. A refined, rapid and reproducible high resolution melt (HRM-based method suitable for quantification of global LINE-1 repetitive element methylation

    Directory of Open Access Journals (Sweden)

    Tse M Yat

    2011-12-01

    Full Text Available Abstract Background The methylation of DNA is recognized as a key mechanism in the regulation of genomic stability and evidence for its role in the development of cancer is accumulating. LINE-1 methylation status represents a surrogate measure of genome-wide methylation. Findings Using high resolution melt (HRM curve analysis technology, we have established an in-tube assay that is linear (r > 0.9986 with a high amplification efficiency (90-105%, capable of discriminating between partcipant samples with small differences in methylation, and suitable for quantifying a wide range of LINE-1 methylation levels (0-100%--including the biologically relevant range of 50-90% expected in human DNA. We have optimized this procedure to perform using 2 μg of starting DNA and 2 ng of bisulfite-converted DNA for each PCR reaction. Intra- and inter-assay coefficients of variation were 1.44% and 0.49%, respectively, supporting the high reproducibility and precision of this approach. Conclusions In summary, this is a completely linear, quantitative HRM PCR method developed for the measurement of LINE-1 methylation. This cost-efficient, refined and reproducible assay can be performed using minimal amounts of starting DNA. These features make our assay suitable for high throughput analysis of multiple samples from large population-based studies.

  10. High-resolution melt and morphological analyses of mealybugs (Hemiptera: Pseudococcidae) from cacao: tools for the control of Cacao swollen shoot virus spread.

    Science.gov (United States)

    Wetten, Andy; Campbell, Colin; Allainguillaume, Joël

    2016-03-01

    Mealybugs (Hemiptera: Coccoidea: Pseudococcidae) are key vectors of badnaviruses, including Cacao swollen shoot virus (CSSV), the most damaging virus affecting cacao (Theobroma cacao L.). The effectiveness of mealybugs as virus vectors is species dependent, and it is therefore vital that CSSV resistance breeding programmes in cacao incorporate accurate mealybug identification. In this work, the efficacy of a CO1-based DNA barcoding approach to species identification was evaluated by screening a range of mealybugs collected from cacao in seven countries. Morphologically similar adult females were characterised by scanning electron microscopy, and then, following DNA extraction, were screened with CO1 barcoding markers. A high degree of CO1 sequence homology was observed for all 11 individual haplotypes, including those accessions from distinct geographical regions. This has allowed the design of a high-resolution melt (HRM) assay capable of rapid identification of the commonly encountered mealybug pests of cacao. HRM analysis readily differentiated between mealybug pests of cacao that cannot necessarily be identified by conventional morphological analysis. This new approach, therefore, has potential to facilitate breeding for resistance to CSSV and other mealybug-transmitted diseases. © 2015 Society of Chemical Industry.

  11. Validation of methylation-sensitive high-resolution melting (MS-HRM) for the detection of stool DNA methylation in colorectal neoplasms.

    Science.gov (United States)

    Xiao, Zhujun; Li, Bingsheng; Wang, Guozhen; Zhu, Weisi; Wang, Zhongqiu; Lin, Jinfeng; Xu, Angao; Wang, Xinying

    2014-04-20

    Methylation-sensitive high-resolution melting (MS-HRM) is a new technique for assaying DNA methylation, but its feasibility for assaying stool in patients with colorectal cancer (CRC) is unknown. First, the MS-HRM and methylation-specific PCR (MSP) detection limits were tested. Second, the methylation statuses of SFRP2 and VIM were analyzed in stool samples by MS-HRM, and in matching tumor and normal colon tissues via bisulfite sequencing PCR (BSP). Third, a case-control study evaluated the diagnostic sensitivity and specificity of MS-HRM relative to results obtained with MSP and the fecal immunochemical test (FIT). Finally, the linearity and reproducibility of MS-HRM were assessed. The detection limits of MS-HRM and MSP were 1% and 5%, respectively. The diagnostic sensitivities of MS-HRM (87.3%, 55/63) in stool and BSP in matching tumor tissue (92.1%, 58/63) were highly consistent (κ=0.744). The MS-HRM assay detected 92.5% (37/40) methylation in CRCs, 94.4% (34/36) in advanced adenomas, and 8.8% (5/57) in normal controls. The results of MS-HRM analysis were stable and reliable and showed fairly good linearity for both SFRP2 (PHRM shows potential for CRC screening. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Use of a high resolution melting (HRM) assay to compare gag, pol, and env diversity in adults with different stages of HIV infection.

    Science.gov (United States)

    Cousins, Matthew M; Laeyendecker, Oliver; Beauchamp, Geetha; Brookmeyer, Ronald; Towler, William I; Hudelson, Sarah E; Khaki, Leila; Koblin, Beryl; Chesney, Margaret; Moore, Richard D; Kelen, Gabor D; Coates, Thomas; Celum, Connie; Buchbinder, Susan P; Seage, George R; Quinn, Thomas C; Donnell, Deborah; Eshleman, Susan H

    2011-01-01

    Cross-sectional assessment of HIV incidence relies on laboratory methods to discriminate between recent and non-recent HIV infection. Because HIV diversifies over time in infected individuals, HIV diversity may serve as a biomarker for assessing HIV incidence. We used a high resolution melting (HRM) diversity assay to compare HIV diversity in adults with different stages of HIV infection. This assay provides a single numeric HRM score that reflects the level of genetic diversity of HIV in a sample from an infected individual. HIV diversity was measured in 203 adults: 20 with acute HIV infection (RNA positive, antibody negative), 116 with recent HIV infection (tested a median of 189 days after a previous negative HIV test, range 14-540 days), and 67 with non-recent HIV infection (HIV infected >2 years). HRM scores were generated for two regions in gag, one region in pol, and three regions in env. Median HRM scores were higher in non-recent infection than in recent infection for all six regions tested. In multivariate models, higher HRM scores in three of the six regions were independently associated with non-recent HIV infection. The HRM diversity assay provides a simple, scalable method for measuring HIV diversity. HRM scores, which reflect the genetic diversity in a viral population, may be useful biomarkers for evaluation of HIV incidence, particularly if multiple regions of the HIV genome are examined.

  13. High-resolution melting (HRM) assay for the detection of recurrent BRCA1/BRCA2 germline mutations in Tunisian breast/ovarian cancer families.

    Science.gov (United States)

    Riahi, Aouatef; Kharrat, Maher; Lariani, Imen; Chaabouni-Bouhamed, Habiba

    2014-12-01

    Germline deleterious mutations in the BRCA1/BRCA2 genes are associated with an increased risk for the development of breast and ovarian cancer. Given the large size of these genes the detection of such mutations represents a considerable technical challenge. Therefore, the development of cost-effective and rapid methods to identify these mutations became a necessity. High resolution melting analysis (HRM) is a rapid and efficient technique extensively employed as high-throughput mutation scanning method. The purpose of our study was to assess the specificity and sensitivity of HRM for BRCA1 and BRCA2 genes scanning. As a first step we estimate the ability of HRM for detection mutations in a set of 21 heterozygous samples harboring 8 different known BRCA1/BRCA2 variations, all samples had been preliminarily investigated by direct sequencing, and then we performed a blinded analysis by HRM in a set of 68 further sporadic samples of unknown genotype. All tested heterozygous BRCA1/BRCA2 variants were easily identified. However the HRM assay revealed further alteration that we initially had not searched (one unclassified variant). Furthermore, sequencing confirmed all the HRM detected mutations in the set of unknown samples, including homozygous changes, indicating that in this cohort, with the optimized assays, the mutations detections sensitivity and specificity were 100 %. HRM is a simple, rapid and efficient scanning method for known and unknown BRCA1/BRCA2 germline mutations. Consequently the method will allow for the economical screening of recurrent mutations in Tunisian population.

  14. Taxonomic Identification of Mediterranean Pines and Their Hybrids Based on the High Resolution Melting (HRM) and trnL Approaches: From Cytoplasmic Inheritance to Timber Tracing

    Science.gov (United States)

    Ganopoulos, Ioannis; Aravanopoulos, Filippos; Madesis, Panagiotis; Pasentsis, Konstantinos; Bosmali, Irene; Ouzounis, Christos; Tsaftaris, Athanasios

    2013-01-01

    Fast and accurate detection of plant species and their hybrids using molecular tools will facilitate the assessment and monitoring of local biodiversity in an era of climate and environmental change. Herein, we evaluate the utility of the plastid trnL marker for species identification applied to Mediterranean pines (Pinus spp.). Our results indicate that trnL is a very sensitive marker for delimiting species biodiversity. Furthermore, High Resolution Melting (HRM) analysis was exploited as a molecular fingerprint for fast and accurate discrimination of Pinus spp. DNA sequence variants. The trnL approach and the HRM analyses were extended to wood samples of two species (Pinus nigra and Pinus sylvestris) with excellent results, congruent to those obtained using leaf tissue. Both analyses demonstrate that hybrids from the P. brutia (maternal parent) × P. halepensis (paternal parent) cross, exhibit the P. halepensis profile, confirming paternal plastid inheritance in Group Halepensis pines. Our study indicates that a single one-step reaction method and DNA marker are sufficient for the identification of Mediterranean pines, their hybrids and the origin of pine wood. Furthermore, our results underline the potential for certain DNA regions to be used as novel biological information markers combined with existing morphological characters and suggest a relatively reliable and open taxonomic system that can link DNA variation to phenotype-based species or hybrid assignment status and direct taxa identification from recalcitrant tissues such as wood samples. PMID:23577179

  15. High-resolution melting (HRM) re-analysis of a polyposis patients cohort reveals previously undetected heterozygous and mosaic APC gene mutations.

    Science.gov (United States)

    Out, Astrid A; van Minderhout, Ivonne J H M; van der Stoep, Nienke; van Bommel, Lysette S R; Kluijt, Irma; Aalfs, Cora; Voorendt, Marsha; Vossen, Rolf H A M; Nielsen, Maartje; Vasen, Hans F A; Morreau, Hans; Devilee, Peter; Tops, Carli M J; Hes, Frederik J

    2015-06-01

    Familial adenomatous polyposis is most frequently caused by pathogenic variants in either the APC gene or the MUTYH gene. The detection rate of pathogenic variants depends on the severity of the phenotype and sensitivity of the screening method, including sensitivity for mosaic variants. For 171 patients with multiple colorectal polyps without previously detectable pathogenic variant, APC was reanalyzed in leukocyte DNA by one uniform technique: high-resolution melting (HRM) analysis. Serial dilution of heterozygous DNA resulted in a lowest detectable allelic fraction of 6% for the majority of variants. HRM analysis and subsequent sequencing detected pathogenic fully heterozygous APC variants in 10 (6%) of the patients and pathogenic mosaic variants in 2 (1%). All these variants were previously missed by various conventional scanning methods. In parallel, HRM APC scanning was applied to DNA isolated from polyp tissue of two additional patients with apparently sporadic polyposis and without detectable pathogenic APC variant in leukocyte DNA. In both patients a pathogenic mosaic APC variant was present in multiple polyps. The detection of pathogenic APC variants in 7% of the patients, including mosaics, illustrates the usefulness of a complete APC gene reanalysis of previously tested patients, by a supplementary scanning method. HRM is a sensitive and fast pre-screening method for reliable detection of heterozygous and mosaic variants, which can be applied to leukocyte and polyp derived DNA.

  16. ITS2 barcoding DNA region combined with high resolution melting (HRM) analysis of Hyoscyami Semen, the mature seed of Hyoscyamus niger.

    Science.gov (United States)

    Xiong, Chao; Hu, Zhi-Gang; Tu, Yuan; Liu, He-Gang; Wang, Ping; Zhao, Ming-Ming; SHIi, Yu-Hua; Wu, Lan; Sun, Wei; Chen, Shi-Lin

    2016-12-01

    Hyoscyami Semen, the mature dried seed of Hyoscyamus niger L., has long been used as a traditional Chinese medicine to treat human diseases. Hyoscyami Semen is found in local markets in China. In markets, sellers and buyers commonly inadvertently mix the seeds of H. niger with the seeds of related species such as Hygrophila salicifolia (Vahl) Nees, Astragalus complanatus R. Br., Cuscuta australis R. Br., Cuscuta chinensis Lam., and Impatiens balsamina L. because of their similar morphologies or similar names. Thus, developing a reliable method for discriminating H. niger seeds from its adulterants is necessary to reduce confusion and ensure the safe use of Hyoscyami Semen. The present study was designed to evaluate the efficiency of high-resolution melting analysis combined with DNA barcoding (Bar-HRM) with internal transcribed spacer 2 to discriminate H. niger. Our results show that Bar-HRM successfully identified the adulterants and detected the proportion of H. niger DNA extract within an admixture. In particular, HRM detected H. niger DNA extract in A. complanatus DNA extract at concentrations as low as 1%. In conclusion, the Bar-HRM method developed in the present study for authenticating H. niger is rapid and cost-effective. It can be used in the future to guarantee the purity of Hyoscyami Semen for the clinical use. Copyright © 2016 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  17. Rapid Identification of Echinococcus granulosus and E. canadensis Using High-Resolution Melting (HRM) Analysis by Focusing on a Single Nucleotide Polymorphism.

    Science.gov (United States)

    Safa, Ahmad Hosseini; Harandi, Majid Fasihi; Tajaddini, Mohammadhasan; Rostami-Nejad, Mohammad; Mohtashami-Pour, Mehdi; Pestehchian, Nader

    2016-07-22

    High-resolution melting (HRM) is a reliable and sensitive scanning method to detect variation in DNA sequences. We used this method to better understand the epidemiology and transmission of Echinococcus granulosus. We tested the use of HRM to discriminate the genotypes of E. granulosus and E. canadensis. One hundred forty-one hydatid cysts were collected from slaughtered animals in different parts of Isfahan-Iran in 2013. After DNA extraction, the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was amplified using PCR coupled with the HRM curve. The result of HRM analysis using partial the sequences of cox1 gene revealed that 93, 35, and 2 isolates were identified as G1, G3, and G6 genotypes, respectively. A single nucleotide polymorphism (SNP) was found in locus 9867 of the cox1 gene. This is a critical locus for the differentiation between the G6 and G7 genotypes. In the phylogenic tree, the sample with a SNP was located between the G6 and G7 genotypes, which suggest that this isolate has a G6/G7 genotype. The HRM analysis developed in the present study provides a powerful technique for molecular and epidemiological studies on echinococcosis in humans and animals.

  18. Detection and characterization of Leishmania (Leishmania and Leishmania (Viannia by SYBR green-based real-time PCR and high resolution melt analysis targeting kinetoplast minicircle DNA.

    Directory of Open Access Journals (Sweden)

    Marcello Ceccarelli

    Full Text Available Leishmaniasis is a neglected disease with a broad clinical spectrum which includes asymptomatic infection. A thorough diagnosis, able to distinguish and quantify Leishmania parasites in a clinical sample, constitutes a key step in choosing an appropriate therapy, making an accurate prognosis and performing epidemiological studies. Several molecular techniques have been shown to be effective in the diagnosis of leishmaniasis. In particular, a number of PCR methods have been developed on various target DNA sequences including kinetoplast minicircle constant regions. The first aim of this study was to develop a SYBR green-based qPCR assay for Leishmania (Leishmania infantum detection and quantification, using kinetoplast minicircle constant region as target. To this end, two assays were compared: the first used previously published primer pairs (qPCR1, whereas the second used a nested primer pairs generating a shorter PCR product (qPCR2. The second aim of this study was to evaluate the possibility to discriminate among subgenera Leishmania (Leishmania and Leishmania (Viannia using the qPCR2 assay followed by melting or High Resolution Melt (HRM analysis. Both assays used in this study showed good sensitivity and specificity, and a good correlation with standard IFAT methods in 62 canine clinical samples. However, the qPCR2 assay allowed to discriminate between Leishmania (Leishmania and Leishmania (Viannia subgenera through melting or HRM analysis. In addition to developing assays, we investigated the number and genetic variability of kinetoplast minicircles in the Leishmania (L. infantum WHO international reference strain (MHOM/TN/80/IPT1, highlighting the presence of minicircle subclasses and sequence heterogeneity. Specifically, the kinetoplast minicircle number per cell was estimated to be 26,566±1,192, while the subclass of minicircles amplifiable by qPCR2 was estimated to be 1,263±115. This heterogeneity, also observed in canine clinical

  19. High-resolution melting analysis of the single nucleotide polymorphism hot-spot region in the rpoB gene as an indicator of reduced susceptibility to rifaximin in Clostridium difficile.

    Science.gov (United States)

    Pecavar, Verena; Blaschitz, Marion; Hufnagl, Peter; Zeinzinger, Josef; Fiedler, Anita; Allerberger, Franz; Maass, Matthias; Indra, Alexander

    2012-06-01

    Clostridium difficile, a Gram-positive, spore-forming, anaerobic bacterium, is the main causative agent of hospital-acquired diarrhoea worldwide. In addition to metronidazole and vancomycin, rifaximin, a rifamycin derivative, is a promising antibiotic for the treatment of recurring C. difficile infections (CDI). However, exposure of C. difficile to this antibiotic has led to the development of rifaximin-resistance due to point mutations in the β-subunit of the RNA polymerase (rpoB) gene. In the present study, 348 C. difficile strains with known PCR-ribotypes were investigated for respective single nucleotide polymorphisms (SNPs) within the proposed rpoB hot-spot region by using high-resolution melting (HRM) analysis. This method allows the detection of SNPs by comparing the altered melting behaviour of dsDNA with that of wild-type DNA. Discrimination between wild-type and mutant strains was enhanced by creating heteroduplexes by mixing sample DNA with wild-type DNA, leading to characteristic melting curve shapes from samples containing SNPs in the respective rpoB section. In the present study, we were able to identify 16 different rpoB sequence-types (ST) by sequencing analysis of a 325 bp fragment. The 16 PCR STs displayed a total of 24 different SNPs. Fifteen of these 24 SNPs were located within the proposed 151 bp SNP hot-spot region, resulting in 11 different HRM curve profiles (CP). Eleven SNPs (seven of which were within the proposed hot-spot region) led to amino acid substitutions associated with reduced susceptibility to rifaximin and 13 SNPs (eight of which were within the hot-spot region) were synonymous. This investigation clearly demonstrates that HRM analysis of the proposed SNP hot-spot region in the rpoB gene of C. difficile is a fast and cost-effective method for the identification of C. difficile samples with reduced susceptibility to rifaximin and even allows simultaneous SNP subtyping of the respective C. difficile isolates.

  20. Detection of sdhB Gene Mutations in SDHI-Resistant Isolates of Botrytis cinerea Using High Resolution Melting (HRM) Analysis.

    Science.gov (United States)

    Samaras, Anastasios; Madesis, Panagiotis; Karaoglanidis, George S

    2016-01-01

    Botrytis cinerea , is a high risk pathogen for fungicide resistance development. Pathogen' resistance to SDHIs is associated with several mutations in sdh gene. The diversity of mutations and their differential effect on cross-resistance patterns among SDHIs and the fitness of resistant strains necessitate the availability of a tool for their rapid identification. This study was initiated to develop and validate a high-resolution melting (HRM) analysis for the identification of P225H/F/L//T, N230I, and H272L/R/Y mutations. Based on the sequence of sdh B subunit of resistant and sensitive isolates, a universal primer pair was designed. The specificity of the HRM analysis primers was verified to ensure against the cross-reaction with other fungal species and its sensitivity was evaluated using concentrations of known amounts of mutant's DNA. The melting curve analysis generated nine distinct curve profiles, enabling the discrimination of all the four mutations located at codon 225, the N230I mutation, the three mutations located in codon 272, and the non-mutated isolates (isolates of wild-type sensitivity). Similar results were obtained when DNA was extracted directly from artificially inoculated strawberry fruit. The method was validated by monitoring the presence of sdh B mutations in samples of naturally infected strawberry fruits and stone fruit rootstock seedling plants showing damping-off symptoms. HRM analysis data were compared with a standard PIRA-PCR technique and an absolute agreement was observed suggesting that in both populations the H272R mutation was the predominant one, while H272Y, N230I, and P225H were detected in lower frequencies. The results of the study suggest that HRM analysis can be a useful tool for sensate, accurate, and rapid identification of several sdh B mutations in B. cinerea and it is expected to contribute in routine fungicide resistance monitoring or assessments of the effectiveness of anti-resistance strategies implemented in

  1. Detection of sdhB gene mutations in SDHI-resistant isolates of Botrytis cinerea using high resolution melting (HRM analysis

    Directory of Open Access Journals (Sweden)

    Anastasios Samaras

    2016-11-01

    Full Text Available Botrytis cinerea, is a high-risk pathogen for fungicide resistance development. Pathogen` resistance to SDHIs is associated with several mutations in sdh gene. The diversity of mutations and their differential effect on cross-resistance patterns among SDHIs and the fitness of resistant strains necessitate the availability of a tool for their rapid identification. This study was initiated to develop and validate a high-resolution melting (HRM analysis for the identification of P225H/F/L//T, N230I and H272L/R/Y mutations. Based on the sequence of sdhB subunit of resistant and sensitive isolates, a universal primer pair was designed. The specificity of the HRM analysis primers was verified to ensure against the cross-reaction with other fungal species and its sensitivity was evaluated using concentrations of known amounts of mutant`s DNA. The melting curve analysis generated nine distinct curve profiles, enabling the discrimination of all the 4 mutations located at codon 225, the N230I mutation, the 3 mutations located in codon 272 and the non mutated isolates (isolates of wild type sensitivity. Similar results were obtained when DNA was extracted directly from artificially inoculated strawberry fruit. The method was validated by monitoring the presence of sdhB mutations in samples of naturally infected strawberry fruits and stone fruit rootstock seedling plants showing damping off symptoms. HRM analysis data were compared with a standard PIRA-PCR technique and an absolute agreement was observed suggesting that in both populations the H272R mutation was the predominant one, while H272Y, N230I and P225H were detected in lower frequencies. The results of the study suggest that HRM analysis can be a useful tool for sensate, accurate and rapid identification of several sdhB mutations in B. cinerea and it is expected to contribute in routine fungicide resistance monitoring or assessments of the effectiveness of antiresistance strategies implemented in

  2. Evaluation of the Capacity of PCR and High-Resolution Melt Curve Analysis for Identification of Mixed Infection with Mycoplasma gallisepticum Strains.

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    Seyed A Ghorashi

    Full Text Available Pathogenicity and presentation of Mycoplasma gallisepticum (MG infection may differ from one strain to another and this may have implications on control measures. Infection of individual birds with more than one MG strain has been reported. A PCR followed by high resolution melt (HRM curve analysis has been developed in our laboratory and routinely used for detection and differentiation of MG strains. However the potential of this test for identification of MG strains in a mixed specimen has not been evaluated. In the present study, the capability of PCR-HRM curve analysis technique, targeting vlhA and pvpA genes was assessed for identification of individual MG strains in a mixed population. Different DNA ratios of two MG strains from 1 to 10(-4 ng were tested with some generated conventional and normalized curves distinct from those of individual strains alone. Using genotype confidence percentages (GCP generated from HRM curve analysis, it was found that vlhA PCR-HRM was more consistent than pvpA PCR-HRM for the detection of MG ts-11 vaccine strain mixed with any of the MG strains 6/85, F, S6 or a field isolate. The potential of vlhA PCR-HRM to detect mixed MG strains in a specimen was found to be primarily dependent on quantity and proportion of the target DNAs in the mixture. This is the first study examining the capacity of PCR-HRM technique for identification of individual MG strains in a mixed strain population.

  3. High-resolution melting PCR assay, applicable for diagnostics and screening studies, allowing detection and differentiation of several Babesia spp. infecting humans and animals.

    Science.gov (United States)

    Rozej-Bielicka, Wioletta; Masny, Aleksander; Golab, Elzbieta

    2017-10-01

    The goal of the study was to design a single tube PCR test for detection and differentiation of Babesia species in DNA samples obtained from diverse biological materials. A multiplex, single tube PCR test was designed for amplification of approximately 400 bp region of the Babesia 18S rRNA gene. Universal primers were designed to match DNA of multiple Babesia spp. and to have low levels of similarity to DNA sequences of other intracellular protozoa and Babesia hosts. The PCR products amplified from Babesia DNA isolated from human, dog, rodent, deer, and tick samples were subjected to high-resolution melting analysis for Babesia species identification. The designed test allowed detection and differentiation of four Babesia species, three zoonotic (B. microti, B. divergens, B. venatorum) and one that is generally not considered zoonotic-Babesia canis. Both detection and identification of all four species were possible based on the HRM curves of the PCR products in samples obtained from the following: humans, dogs, rodents, and ticks. No cross-reactivity with DNA of Babesia hosts or Plasmodium falciparum and Toxoplasma gondii was observed. The lack of cross-reactivity with P. falciparum DNA might allow using the assay in endemic malaria areas. The designed assay is the first PCR-based test for detection and differentiation of several Babesia spp. of medical and veterinary importance, in a single tube reaction. The results of the study show that the designed assay for Babesia detection and identification could be a practical and inexpensive tool for diagnostics and screening studies of diverse biological materials.

  4. CtGEM typing: Discrimination of Chlamydia trachomatis ocular and urogenital strains and major evolutionary lineages by high resolution melting analysis of two amplified DNA fragments.

    Science.gov (United States)

    Giffard, Philip M; Andersson, Patiyan; Wilson, Judith; Buckley, Cameron; Lilliebridge, Rachael; Harris, Tegan M; Kleinecke, Mariana; O'Grady, Kerry-Ann F; Huston, Wilhelmina M; Lambert, Stephen B; Whiley, David M; Holt, Deborah C

    2018-01-01

    Chlamydia trachomatis infects the urogenital tract (UGT) and eyes. Anatomical tropism is correlated with variation in the major outer membrane protein encoded by ompA. Strains possessing the ocular ompA variants A, B, Ba and C are typically found within the phylogenetically coherent "classical ocular lineage". However, variants B, Ba and C have also been found within three distinct strains in Australia, all associated with ocular disease in children and outside the classical ocular lineage. CtGEM genotyping is a method for detecting and discriminating ocular strains and also the major phylogenetic lineages. The rationale was facilitation of surveillance to inform responses to C. trachomatis detection in UGT specimens from young children. CtGEM typing is based on high resolution melting analysis (HRMA) of two PCR amplified fragments with high combinatorial resolving power, as defined by computerised comparison of 65 whole genomes. One fragment is from the hypothetical gene defined by Jali-1891 in the C. trachomatis B_Jali20 genome, while the other is from ompA. Twenty combinatorial CtGEM types have been shown to exist, and these encompass unique genotypes for all known ocular strains, and also delineate the TI and T2 major phylogenetic lineages, identify LGV strains and provide additional resolution beyond this. CtGEM typing and Sanger sequencing were compared with 42 C. trachomatis positive clinical specimens, and there were no disjunctions. CtGEM typing is a highly efficient method designed and tested using large scale comparative genomics. It divides C. trachomatis into clinically and biologically meaningful groups, and may have broad application in surveillance.

  5. Establishment of a simple and rapid identification method for Listeria spp. by using high-resolution melting analysis, and its application in food industry.

    Science.gov (United States)

    Ohshima, Chihiro; Takahashi, Hajime; Phraephaisarn, Chirapiphat; Vesaratchavest, Mongkol; Keeratipibul, Suwimon; Kuda, Takashi; Kimura, Bon

    2014-01-01

    Listeria monocytogenes is the causative bacteria of listeriosis, which has a higher mortality rate than that of other causes of food poisoning. Listeria spp., of which L. monocytogenes is a member, have been isolated from food and manufacturing environments. Several methods have been published for identifying Listeria spp.; however, many of the methods cannot identify newly categorized Listeria spp. Additionally, they are often not suitable for the food industry, owing to their complexity, cost, or time consumption. Recently, high-resolution melting analysis (HRMA), which exploits DNA-sequence differences, has received attention as a simple and quick genomic typing method. In the present study, a new method for the simple, rapid, and low-cost identification of Listeria spp. has been presented using the genes rarA and ldh as targets for HRMA. DNA sequences of 9 Listeria species were first compared, and polymorphisms were identified for each species for primer design. Species specificity of each HRM curve pattern was estimated using type strains of all the species. Among the 9 species, 7 were identified by HRMA using rarA gene, including 3 new species. The remaining 2 species were identified by HRMA of ldh gene. The newly developed HRMA method was then used to assess Listeria isolates from the food industry, and the method efficiency was compared to that of identification by 16S rDNA sequence analysis. The 2 methods were in coherence for 92.6% of the samples, demonstrating the high accuracy of HRMA. The time required for identifying Listeria spp. was substantially low, and the process was considerably simplified, providing a useful and precise method for processing multiple samples per day. Our newly developed method for identifying Listeria spp. is highly valuable; its use is not limited to the food industry, and it can be used for the isolates from the natural environment.

  6. Towards a pathogenic Escherichia coli detection platform using multiplex SYBR®Green Real-time PCR methods and high resolution melting analysis.

    Directory of Open Access Journals (Sweden)

    Dafni-Maria Kagkli

    Full Text Available Escherichia coli is a group of bacteria which has raised a lot of safety concerns in recent years. Five major intestinal pathogenic groups have been recognized amongst which the verocytotoxin or shiga-toxin (stx1 and/or stx2 producing E. coli (VTEC or STEC respectively have received a lot of attention recently. Indeed, due to the high number of outbreaks related to VTEC strains, the European Food Safety Authority (EFSA has requested the monitoring of the "top-five" serogroups (O26, O103, O111, O145 and O157 most often encountered in food borne diseases and addressed the need for validated VTEC detection methods. Here we report the development of a set of intercalating dye Real-time PCR methods capable of rapidly detecting the presence of the toxin genes together with intimin (eae in the case of VTEC, or aggregative protein (aggR, in the case of the O104:H4 strain responsible for the outbreak in Germany in 2011. All reactions were optimized to perform at the same annealing temperature permitting the multiplex application in order to minimize the need of material and to allow for high-throughput analysis. In addition, High Resolution Melting (HRM analysis allowing the discrimination among strains possessing similar virulence traits was established. The development, application to food samples and the flexibility in use of the methods are thoroughly discussed. Together, these Real-time PCR methods facilitate the detection of VTEC in a new highly efficient way and could represent the basis for developing a simple pathogenic E. coli platform.

  7. Identification and Evaluation of Single-Nucleotide Polymorphisms in Allotetraploid Peanut (Arachis hypogaea L.) Based on Amplicon Sequencing Combined with High Resolution Melting (HRM) Analysis.

    Science.gov (United States)

    Hong, Yanbin; Pandey, Manish K; Liu, Ying; Chen, Xiaoping; Liu, Hong; Varshney, Rajeev K; Liang, Xuanqiang; Huang, Shangzhi

    2015-01-01

    The cultivated peanut (Arachis hypogaea L.) is an allotetraploid (AABB) species derived from the A-genome (Arachis duranensis) and B-genome (Arachis ipaensis) progenitors. Presence of two versions of a DNA sequence based on the two progenitor genomes poses a serious technical and analytical problem during single nucleotide polymorphism (SNP) marker identification and analysis. In this context, we have analyzed 200 amplicons derived from expressed sequence tags (ESTs) and genome survey sequences (GSS) to identify SNPs in a panel of genotypes consisting of 12 cultivated peanut varieties and two diploid progenitors representing the ancestral genomes. A total of 18 EST-SNPs and 44 genomic-SNPs were identified in 12 peanut varieties by aligning the sequence of A. hypogaea with diploid progenitors. The average frequency of sequence polymorphism was higher for genomic-SNPs than the EST-SNPs with one genomic-SNP every 1011 bp as compared to one EST-SNP every 2557 bp. In order to estimate the potential and further applicability of these identified SNPs, 96 peanut varieties were genotyped using high resolution melting (HRM) method. Polymorphism information content (PIC) values for EST-SNPs ranged between 0.021 and 0.413 with a mean of 0.172 in the set of peanut varieties, while genomic-SNPs ranged between 0.080 and 0.478 with a mean of 0.249. Total 33 SNPs were used for polymorphism detection among the parents and 10 selected lines from mapping population Y13Zh (Zhenzhuhei × Yueyou13). Of the total 33 SNPs, nine SNPs showed polymorphism in the mapping population Y13Zh, and seven SNPs were successfully mapped into five linkage groups. Our results showed that SNPs can be identified in allotetraploid peanut with high accuracy through amplicon sequencing and HRM assay. The identified SNPs were very informative and can be used for different genetic and breeding applications in peanut.

  8. Ribo HRM--detection of inter- and intra-species polymorphisms within ribosomal DNA by high resolution melting analysis supported by application of artificial allelic standards.

    Science.gov (United States)

    Masny, Aleksander; Jagiełło, Agata; Płucienniczak, Grażyna; Golab, Elzbieta

    2012-09-01

    Ribo HRM, a single-tube PCR and high resolution melting (HRM) assay for detection of polymorphisms in the large subunit ribosomal DNA expansion segment V, was developed on a Trichinella model. Four Trichinella species: T. spiralis (isolates ISS3 and ISS160), T. nativa (isolates ISS10 and ISS70), T. britovi (isolates ISS2 and ISS392) and T. pseudospiralis (isolates ISS13 and ISS1348) were genotyped. Cloned allelic variants of the expansion segment V were used as standards to prepare reference HRM curves characteristic for single sequences and mixtures of several cloned sequences imitating allelic composition detected in Trichinella isolates. Using the primer pair Tsr1 and Trich1bi, it was possible to amplify a fragment of the ESV and detect PCR products obtained from the genomic DNA of pools of larvae belonging to the four investigated species: T. pseudospiralis, T. spiralis, T. britovi and T. nativa, in a single tube Real-Time PCR reaction. Differences in the shape of the HRM curves of Trichinella isolates suggested the presence of differences between examined isolates of T. nativa, T. britovi and T. pseudospiralis species. No differences were observed between T. spiralis isolates. The presence of polymorphisms within the amplified ESV sequence fragment of T. nativa T. britovi and T. pseudospiralis was confirmed by sequencing of the cloned PCR products. Novel sequences were discovered and deposited in GenBank (GenBank IDs: JN971020-JN971027, JN120902.1, JN120903.1, JN120904.1, JN120906.1, JN120905.1). Screening the ESV region of Trichinella for polymorphism is possible using the genotyping assay Ribo HRM at the current state of its development. The Ribo HRM assay could be useful in phylogenetic studies of the Trichinella genus. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. High resolution melting (HRM) analysis as a new tool for rapid identification of Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum.

    Science.gov (United States)

    Ren, Xingxing; Fu, Ying; Xu, Chenggang; Feng, Zhou; Li, Miao; Zhang, Lina; Zhang, Jianmin; Liao, Ming

    2017-05-01

    Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum represent the most common causative agents of chicken salmonellosis, which result in high mortality and morbidity throughout the world. It is difficult and laborious to discriminate these diseases based on biochemical or phenotypic methods. Herein, we report the development of a single nucleotide polymorphism (SNP) PCR-high resolution melt (PCR-HRM) assay for the detection and discrimination of both S. Pullorum and S. Gallinarun. The gene rfbS, which encodes a factor involved in the biosynthesis of ADP paratose in serogroup D of Salmonella, has been identified as a robust genetic marker for the identification of S. Pullorum and S. Gallinarun based on polymorphisms at positions 237 and 598. Therefore, PCR-HRM analyses were used to characterize this gene. A total of 15 reference and 33 clinical isolates of Salmonella and related Gram-negative bacteria were detected using 2 sets of primers. Our PCR-HRM assay could distinguish S. Pullorum from S. Gallinarun and other strains using the primer pair SP-237F/237R. Similarly, S. Gallinarun could be distinguished from S. Pullorum and other strains using primer set SG-598F/598R. These 2 assays showed high specificity (100%) for both S. Pullorum and S. Gallinarun; the sensitivity of these 2 assays was at least 100-fold greater than that of the allele-specific PCR assay. This present study demonstrated that HRM analysis represents a potent, simple, and economic tool for the rapid, specific, and sensitive detection of S. Pullorum and S. Gallinarun. Our approach also may aid efforts for purification of Avian Salmonella disease. © 2016 Poultry Science Association Inc.

  10. Rapid screening of rpoB and katG mutations in Mycobacterium tuberculosis isolates by high-resolution melting curve analysis

    Directory of Open Access Journals (Sweden)

    M Haeili

    2014-01-01

    Full Text Available Background: Early detection of multidrug-resistant tuberculosis (MDR-TB is essential to prevent its transmission in the community and initiate effective anti-TB treatment regimen. Materials and Methods: High-resolution melting curve (HRM analysis was evaluated for rapid detection of resistance conferring mutations in rpoB and katG genes. We screened 95 Mycobacterium tuberculosis clinical isolates including 20 rifampin resistant (RIF-R, 21 isoniazid resistant (INH-R and 54 fully susceptible (S isolates determined by proportion method of drug susceptibility testing. Nineteen M. tuberculosis isolates with known drug susceptibility genotypes were used as references for the assay validation. The nucleotide sequences of the target regions rpoB and katG genes were determined to investigate the frequency and type of mutations and to confirm HRM results. Results: HRM analysis of a 129-bp fragment of rpoB allowed correct identification of 19 of the 20 phenotypically RIF-R and all RIF-S isolates. All INH-S isolates generated wild-type HRM curves and 18 out of 21 INH-R isolates harboured any mutation in 109-bp fragment of katG exhibited mutant type HRM curves. However, 1 RIF-R and 3 INH-R isolates were falsely identified as susceptible which were confirmed for having no mutation in their target regions by sequencing. The main mutations involved in RIF and INH resistance were found at codons rpoB531 (60% of RIF-R isolates and katG315 (85.7% of INH-R isolates, respectively. Conclusion: HRM was found to be a reliable, rapid and low cost method to characterise drug susceptibility of clinical TB isolates in resource-limited settings.

  11. A simple, high throughput method to locate single copy sequences from Bacterial Artificial Chromosome (BAC libraries using High Resolution Melt analysis

    Directory of Open Access Journals (Sweden)

    Caligari Peter DS

    2010-05-01

    Full Text Available Abstract Background The high-throughput anchoring of genetic markers into contigs is required for many ongoing physical mapping projects. Multidimentional BAC pooling strategies for PCR-based screening of large insert libraries is a widely used alternative to high density filter hybridisation of bacterial colonies. To date, concerns over reliability have led most if not all groups engaged in high throughput physical mapping projects to favour BAC DNA isolation prior to amplification by conventional PCR. Results Here, we report the first combined use of Multiplex Tandem PCR (MT-PCR and High Resolution Melt (HRM analysis on bacterial stocks of BAC library superpools as a means of rapidly anchoring markers to BAC colonies and thereby to integrate genetic and physical maps. We exemplify the approach using a BAC library of the model plant Arabidopsis thaliana. Super pools of twenty five 384-well plates and two-dimension matrix pools of the BAC library were prepared for marker screening. The entire procedure only requires around 3 h to anchor one marker. Conclusions A pre-amplification step during MT-PCR allows high multiplexing and increases the sensitivity and reliability of subsequent HRM discrimination. This simple gel-free protocol is more reliable, faster and far less costly than conventional PCR screening. The option to screen in parallel 3 genetic markers in one MT-PCR-HRM reaction using templates from directly pooled bacterial stocks of BAC-containing bacteria further reduces time for anchoring markers in physical maps of species with large genomes.

  12. High resolution, high sensitivity imaging and analysis of minerals and inclusions (fluid and melt) using the new CSIRO-GEMOC nuclear microprobe

    International Nuclear Information System (INIS)

    Ryan, C.G.; McInnes, B.M.; Van Achterbergh, E.; Williams, P.J.; Dong, G.; Zaw, K.

    1999-01-01

    Full text: The new CSIRO-GEMOC Nuclear Microprobe (NMP) The instrument was designed specifically for minerals analysis and imaging and to achieve ppm to sub-ppm sensitivity at a spatial resolution of 1-2 μm using X-rays and y-rays induced by MeV energy ion beams. The key feature of the design is a unique magnetic quadrupole quintuplet ion focussing system that combines high current with high spatial resolution (Ryan et al., 1999). These design goals have been achieved or exceeded. On the first day of operation, a spot-size of 1.3 μm was obtained at a beam current of 0.5 nA, suitable for fluid inclusion analysis and imaging. The spot-size grows to just 1.8 μm at 10 nA (3 MeV protons), ideal for mineralogical samples with detection limits down to 0.2 ppm achieved in quantitative, high resolution, trace element images. Applications of the NMP include: research into ore deposit processes through trace element geochemistry, mineralogy and fluid inclusion analysis of ancient deposits and active sea-floor environments, ore characterization, and fundamental studies of mantle processes and extraterrestrial material. Quantitative True Elemental Imaging Dynamic Analysis is a method for projecting quantitative major and trace element images from proton-induced X-ray emission (PIXE) data obtained using the NMP (Ryan et al., 1995). The method un-mixes full elemental spectral signatures to produce quantitative images that can be directly interrogated for the concentrations of all elements in selected areas or line projections, etc. Fluid Inclusion Analysis and Imaging The analysis of fluids trapped as fluid inclusions in minerals holds the key to understanding ore metal pathways and ore formation processes. PIXE analysis using the NMP provides a direct non-destructive method to determine the composition of these trapped fluids with detection limits down to 20 ppm. However, some PIXE results have been controversial, such as the strong partitioning of Cu into the vapour phase (e

  13. qPCR-High resolution melt analysis for drug susceptibility testing of Mycobacterium leprae directly from clinical specimens of leprosy patients.

    Science.gov (United States)

    Araujo, Sergio; Goulart, Luiz Ricardo; Truman, Richard W; Goulart, Isabela Maria B; Vissa, Varalakshmi; Li, Wei; Matsuoka, Masanori; Suffys, Philip; Fontes, Amanda B; Rosa, Patricia S; Scollard, David M; Williams, Diana L

    2017-06-01

    Real-Time PCR-High Resolution Melting (qPCR-HRM) analysis has been recently described for rapid drug susceptibility testing (DST) of Mycobacterium leprae. The purpose of the current study was to further evaluate the validity, reliability, and accuracy of this assay for M. leprae DST in clinical specimens. The specificity and sensitivity for determining the presence and susceptibility of M. leprae to dapsone based on the folP1 drug resistance determining region (DRDR), rifampin (rpoB DRDR) and ofloxacin (gyrA DRDR) was evaluated using 211 clinical specimens from leprosy patients, including 156 multibacillary (MB) and 55 paucibacillary (PB) cases. When comparing the results of qPCR-HRM DST and PCR/direct DNA sequencing, 100% concordance was obtained. The effects of in-house phenol/chloroform extraction versus column-based DNA purification protocols, and that of storage and fixation protocols of specimens for qPCR-HRM DST, were also evaluated. qPCR-HRM results for all DRDR gene assays (folP1, rpoB, and gyrA) were obtained from both MB (154/156; 98.7%) and PB (35/55; 63.3%) patients. All PCR negative specimens were from patients with low numbers of bacilli enumerated by an M. leprae-specific qPCR. We observed that frozen and formalin-fixed paraffin embedded (FFPE) tissues or archival Fite's stained slides were suitable for HRM analysis. Among 20 mycobacterial and other skin bacterial species tested, only M. lepromatosis, highly related to M. leprae, generated amplicons in the qPCR-HRM DST assay for folP1 and rpoB DRDR targets. Both DNA purification protocols tested were efficient in recovering DNA suitable for HRM analysis. However, 3% of clinical specimens purified using the phenol/chloroform DNA purification protocol gave false drug resistant data. DNA obtained from freshly frozen (n = 172), formalin-fixed paraffin embedded (FFPE) tissues (n = 36) or archival Fite's stained slides (n = 3) were suitable for qPCR-HRM DST analysis. The HRM-based assay was also able to

  14. A multi-step strategy to obtain crystals of the dengue virus RNA-dependent RNA polymerase that diffract to high resolution

    International Nuclear Information System (INIS)

    Yap, Thai Leong; Chen, Yen Liang; Xu, Ting; Wen, Daying; Vasudevan, Subhash G.; Lescar, Julien

    2007-01-01

    Crystals of the RNA-dependent RNA polymerase catalytic domain from the dengue virus NS5 protein have been obtained using a strategy that included expression screening of naturally occurring serotype variants of the protein, the addition of divalent metal ions and crystal dehydration. These crystals diffract to 1.85 Å resolution and are thus suitable for a structure-based drug-design program. Dengue virus, a member of the Flaviviridae genus, causes dengue fever, an important emerging disease with several million infections occurring annually for which no effective therapy exists. The viral RNA-dependent RNA polymerase NS5 plays an important role in virus replication and represents an interesting target for the development of specific antiviral compounds. Crystals that diffract to 1.85 Å resolution that are suitable for three-dimensional structure determination and thus for a structure-based drug-design program have been obtained using a strategy that included expression screening of naturally occurring serotype variants of the protein, the addition of divalent metal ions and crystal dehydration

  15. A multi-step strategy to obtain crystals of the dengue virus RNA-dependent RNA polymerase that diffract to high resolution

    Energy Technology Data Exchange (ETDEWEB)

    Yap, Thai Leong [Novartis Institute for Tropical Diseases, 10 Biopolis Road, Chromos Building, Singapore 138670 (Singapore); School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551 (Singapore); Chen, Yen Liang; Xu, Ting; Wen, Daying; Vasudevan, Subhash G. [Novartis Institute for Tropical Diseases, 10 Biopolis Road, Chromos Building, Singapore 138670 (Singapore); Lescar, Julien, E-mail: julien@ntu.edu.sg [Novartis Institute for Tropical Diseases, 10 Biopolis Road, Chromos Building, Singapore 138670 (Singapore); School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551 (Singapore)

    2007-02-01

    Crystals of the RNA-dependent RNA polymerase catalytic domain from the dengue virus NS5 protein have been obtained using a strategy that included expression screening of naturally occurring serotype variants of the protein, the addition of divalent metal ions and crystal dehydration. These crystals diffract to 1.85 Å resolution and are thus suitable for a structure-based drug-design program. Dengue virus, a member of the Flaviviridae genus, causes dengue fever, an important emerging disease with several million infections occurring annually for which no effective therapy exists. The viral RNA-dependent RNA polymerase NS5 plays an important role in virus replication and represents an interesting target for the development of specific antiviral compounds. Crystals that diffract to 1.85 Å resolution that are suitable for three-dimensional structure determination and thus for a structure-based drug-design program have been obtained using a strategy that included expression screening of naturally occurring serotype variants of the protein, the addition of divalent metal ions and crystal dehydration.

  16. Real-time RT-PCR high-resolution melting curve analysis and multiplex RT-PCR to detect and differentiate grapevine leafroll-associated associated virus 3 variant groups I, II, III and VI

    Directory of Open Access Journals (Sweden)

    Bester Rachelle

    2012-09-01

    Full Text Available Abstract Background Grapevine leafroll-associated virus 3 (GLRaV-3 is the main contributing agent of leafroll disease worldwide. Four of the six GLRaV-3 variant groups known have been found in South Africa, but their individual contribution to leafroll disease is unknown. In order to study the pathogenesis of leafroll disease, a sensitive and accurate diagnostic assay is required that can detect different variant groups of GLRaV-3. Methods In this study, a one-step real-time RT-PCR, followed by high-resolution melting (HRM curve analysis for the simultaneous detection and identification of GLRaV-3 variants of groups I, II, III and VI, was developed. A melting point confidence interval for each variant group was calculated to include at least 90% of all melting points observed. A multiplex RT-PCR protocol was developed to these four variant groups in order to assess the efficacy of the real-time RT-PCR HRM assay. Results A universal primer set for GLRaV-3 targeting the heat shock protein 70 homologue (Hsp70h gene of GLRaV-3 was designed that is able to detect GLRaV-3 variant groups I, II, III and VI and differentiate between them with high-resolution melting curve analysis. The real-time RT-PCR HRM and the multiplex RT-PCR were optimized using 121 GLRaV-3 positive samples. Due to a considerable variation in melting profile observed within each GLRaV-3 group, a confidence interval of above 90% was calculated for each variant group, based on the range and distribution of melting points. The intervals of groups I and II could not be distinguished and a 95% joint confidence interval was calculated for simultaneous detection of group I and II variants. An additional primer pair targeting GLRaV-3 ORF1a was developed that can be used in a subsequent real-time RT-PCR HRM to differentiate between variants of groups I and II. Additionally, the multiplex RT-PCR successfully validated 94.64% of the infections detected with the real-time RT-PCR HRM

  17. A new method for simultaneous detection and discrimination of Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) using real time PCR with high resolution melting (HRM) analysis.

    Science.gov (United States)

    Marin, M S; Quintana, S; Leunda, M R; Recavarren, M; Pagnuco, I; Späth, E; Pérez, S; Odeón, A

    2016-01-01

    Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of experimentally infected animals were studied by real time PCR-HRM. An efficient amplification of real time PCR products was obtained, and a clear melting curve and appropriate melting peaks for both viruses were achieved in the HRM curve analysis for BoHV type identification. BoHV was identified in all of the isolates and clinical samples, and BoHV types were properly differentiated. Furthermore, viral DNA was detected in 12/18 and 7/18 samples from BoHV-1- and BoHV-5-infected calves, respectively. Real time PCR-HRM achieved a higher sensitivity compared with virus isolation or conventional PCR. In this study, HRM was used as a novel procedure. This method provides rapid, sensitive, specific and simultaneous detection of bovine alpha-herpesviruses DNA. Thus, this technique is an excellent tool for diagnosis, research and epidemiological studies of these viruses in cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Development of a two-step high-resolution melting (HRM) analysis for screening sequence variants associated with resistance to the QoIs, benzimidazoles and dicarboximides in airborne inoculum of Botrytis cinerea.

    Science.gov (United States)

    Chatzidimopoulos, Michael; Ganopoulos, Ioannis; Vellios, Evangelos; Madesis, Panagiotis; Tsaftaris, Athanasios; Pappas, Athanassios C

    2014-11-01

    A rapid, high-resolution melting (HRM) analysis protocol was developed to detect sequence variations associated with resistance to the QoIs, benzimidazoles and dicarboximides in Botrytis cinerea airborne inoculum. HRM analysis was applied directly in fungal DNA collected from air samplers with selective medium. Three and five different genotypes were detected and classified according to their melting profiles in BenA and bos1 genes associated with resistance to benzimidazoles and dicarboximides, respectively. The sensitivity of the methodology was evident in the case of the QoIs, where genotypes varying either by a single nucleotide polymorphism or an additional 1205-bp intron were separated accurately with a single pair of primers. The developed two-step protocol was completed in 82 min and showed reduced variation in the melting curves' formation. HRM analysis rapidly detected the major mutations found in greenhouse strains providing accurate data for successfully controlling grey mould. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  19. Use of a high resolution melt real-time polymerase chain reaction (PCR) assay for the environmental monitoring of Vibrio cholerae

    CSIR Research Space (South Africa)

    Le Rouw, Wouter J

    2011-10-01

    Full Text Available ATCC 9207 Shigella sonnei ATCC 9290 Enterococcus faecium ATCC 6569 Enterobacter aerogenes ATCC 13048 Klebsiella pneumoniae Vermaak and Partners Pathologists, S.A. Pseudomonas aeruginosa ATCC 27853 Staphylococcus aureus ATCC 25923 Proteus...

  20. High-resolution melting-curve analysis of ligation-mediated real-time PCR for rapid evaluation of an epidemiological outbreak of extended-spectrum-beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E; Schön, Thomas

    2011-12-01

    Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE.

  1. High-Resolution Melting-Curve Analysis of Ligation-Mediated Real-Time PCR for Rapid Evaluation of an Epidemiological Outbreak of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli ▿

    Science.gov (United States)

    Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E.; Schön, Thomas

    2011-01-01

    Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE. PMID:21956981

  2. High Resolution Elevation Contours

    Data.gov (United States)

    Minnesota Department of Natural Resources — This dataset contains contours generated from high resolution data sources such as LiDAR. Generally speaking this data is 2 foot or less contour interval.

  3. Competitive PCR-High Resolution Melting Analysis (C-PCR-HRMA) for large genomic rearrangements (LGRs) detection: A new approach to assess quantitative status of BRCA1 gene in a reference laboratory.

    Science.gov (United States)

    Minucci, Angelo; De Paolis, Elisa; Concolino, Paola; De Bonis, Maria; Rizza, Roberta; Canu, Giulia; Scaglione, Giovanni Luca; Mignone, Flavio; Scambia, Giovanni; Zuppi, Cecilia; Capoluongo, Ettore

    2017-07-01

    Evaluation of copy number variation (CNV) in BRCA1/2 genes, due to large genomic rearrangements (LGRs), is a mandatory analysis in hereditary breast and ovarian cancers families, if no pathogenic variants are found by sequencing. LGRs cannot be detected by conventional methods and several alternative methods have been developed. Since these approaches are expensive and time consuming, identification of alternative screening methods for LGRs detection is needed in order to reduce and optimize the diagnostic procedure. The aim of this study was to investigate a Competitive PCR-High Resolution Melting Analysis (C-PCR-HRMA) as molecular tool to detect recurrent BRCA1 LGRs. C-PCR-HRMA was performed on exons 3, 14, 18, 19, 20 and 21 of the BRCA1 gene; exons 4, 6 and 7 of the ALB gene were used as reference fragments. This study showed that it is possible to identify recurrent BRCA1 LGRs, by melting peak height ratio between target (BRCA1) and reference (ALB) fragments. Furthermore, we underline that a peculiar amplicon-melting profile is associated to a specific BRCA1 LGR. All C-PCR-HRMA results were confirmed by Multiplex ligation-dependent probe amplification. C-PCR-HRMA has proved to be an innovative, efficient and fast method for BRCA1 LGRs detection. Given the sensitivity, specificity and ease of use, c-PCR-HRMA can be considered an attractive and powerful alternative to other methods for BRCA1 CNVs screening, improving molecular strategies for BRCA testing in the context of Massive Parallel Sequencing. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. High resolution solar observations

    International Nuclear Information System (INIS)

    Title, A.

    1985-01-01

    Currently there is a world-wide effort to develop optical technology required for large diffraction limited telescopes that must operate with high optical fluxes. These developments can be used to significantly improve high resolution solar telescopes both on the ground and in space. When looking at the problem of high resolution observations it is essential to keep in mind that a diffraction limited telescope is an interferometer. Even a 30 cm aperture telescope, which is small for high resolution observations, is a big interferometer. Meter class and above diffraction limited telescopes can be expected to be very unforgiving of inattention to details. Unfortunately, even when an earth based telescope has perfect optics there are still problems with the quality of its optical path. The optical path includes not only the interior of the telescope, but also the immediate interface between the telescope and the atmosphere, and finally the atmosphere itself

  5. Rapid detection of pathological mutations and deletions of the haemoglobin beta gene (HBB) by High Resolution Melting (HRM) analysis and Gene Ratio Analysis Copy Enumeration PCR (GRACE-PCR).

    Science.gov (United States)

    Turner, Andrew; Sasse, Jurgen; Varadi, Aniko

    2016-10-19

    Inherited disorders of haemoglobin are the world's most common genetic diseases, resulting in significant morbidity and mortality. The large number of mutations associated with the haemoglobin beta gene (HBB) makes gene scanning by High Resolution Melting (HRM) PCR an attractive diagnostic approach. However, existing HRM-PCR assays are not able to detect all common point mutations and have only a very limited ability to detect larger gene rearrangements. The aim of the current study was to develop a HBB assay, which can be used as a screening test in highly heterogeneous populations, for detection of both point mutations and larger gene rearrangements. The assay is based on a combination of conventional HRM-PCR and a novel Gene Ratio Analysis Copy Enumeration (GRACE) PCR method. HRM-PCR was extensively optimised, which included the use of an unlabelled probe and incorporation of universal bases into primers to prevent interference from common non-pathological polymorphisms. GRACE-PCR was employed to determine HBB gene copy numbers relative to a reference gene using melt curve analysis to detect rearrangements in the HBB gene. The performance of the assay was evaluated by analysing 410 samples. A total of 44 distinct pathological genotypes were detected. In comparison with reference methods, the assay has a sensitivity of 100 % and a specificity of 98 %. We have developed an assay that detects both point mutations and larger rearrangements of the HBB gene. This assay is quick, sensitive, specific and cost effective making it suitable as an initial screening test that can be used for highly heterogeneous cohorts.

  6. Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM analysis [v1; ref status: indexed, http://f1000r.es/2hj

    Directory of Open Access Journals (Sweden)

    Erin K. Hanson

    2013-12-01

    Full Text Available Positive identification of the nature of biological material present on evidentiary items can be crucial for understanding the circumstances surrounding a crime. However, traditional protein-based methods do not permit the identification of all body fluids and tissues, and thus molecular based strategies for the conclusive identification of all forensically relevant biological fluids and tissues need to be developed. Messenger RNA (mRNA profiling is an example of such a molecular-based approach. Current mRNA body fluid identification assays involve capillary electrophoresis (CE or quantitative RT-PCR (qRT-PCR platforms, each with its own limitations. Both platforms require the use of expensive fluorescently labeled primers or probes. CE-based assays require separate amplification and detection steps thus increasing the analysis time. For qRT-PCR assays, only 3-4 markers can be included in a single reaction since each requires a different fluorescent dye. To simplify mRNA profiling assays, and reduce the time and cost of analysis, we have developed single- and multiplex body fluid High Resolution Melt (HRM assays for the identification of common forensically relevant biological fluids and tissues. The incorporated biomarkers include IL19 (vaginal secretions, IL1F7 (skin, ALAS2 (blood, MMP10 (menstrual blood, HTN3 (saliva and TGM4 (semen.  The HRM assays require only unlabeled PCR primers and a single saturating intercalating fluorescent dye (Eva Green. Each body-fluid-specific marker can easily be identified by the presence of a distinct melt peak. Usually, HRM assays are used to detect variants or isoforms for a single gene target. However, we have uniquely developed duplex and triplex HRM assays to permit the simultaneous detection of multiple targets per reaction. Here we describe the development and initial performance evaluation of the developed HRM assays. The results demonstrate the potential use of HRM assays for rapid, and relatively

  7. Rapid detection of the H275Y oseltamivir resistance mutation in influenza A/H1N1 2009 by single base pair RT-PCR and high-resolution melting.

    Directory of Open Access Journals (Sweden)

    Steven Y C Tong

    Full Text Available We aimed to design a real-time reverse-transcriptase-PCR (rRT-PCR, high-resolution melting (HRM assay to detect the H275Y mutation that confers oseltamivir resistance in influenza A/H1N1 2009 viruses.A novel strategy of amplifying a single base pair, the relevant SNP at position 823 of the neuraminidase gene, was chosen to maintain specificity of the assay. Wildtype and mutant virus were differentiated when using known reference samples of cell-cultured virus. However, when dilutions of these reference samples were assayed, amplification of non-specific primer-dimer was evident and affected the overall melting temperature (T(m of the amplified products. Due to primer-dimer appearance at >30 cycles we found that if the cycle threshold (C(T for a dilution was >30, the HRM assay did not consistently discriminate mutant from wildtype. Where the C(T was 32.98 would have an H275Y assay C(T>30. Analysis of the TaqMan C(T values for 609 consecutive clinical samples predicted that 207 (34% of the samples would result in an HRM assay C(T>30 and therefore not be amenable to the HRM assay.The use of single base pair PCR and HRM can be useful for specifically interrogating SNPs. When applied to H1N1 09, the constraints this placed on primer design resulted in amplification of primer-dimer products. The impact primer-dimer had on HRM curves was adjusted for by plotting T(m against C(T. Although less sensitive than TaqMan assays, the HRM assay can rapidly, and at low cost, screen samples with moderate viral concentrations.

  8. High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) for the characterisation of pathogenic leptospires: intra-serovar divergence, inter-serovar convergence, and evidence of attenuation in Leptospira reference collections.

    Science.gov (United States)

    Tulsiani, S M; Craig, S B; Graham, G C; Cobbold, R C; Dohnt, M F; Burns, M-A; Jansen, C C; Leung, L K-P; Field, H E; Smythe, L D

    2010-07-01

    High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) is a novel technology that has emerged as a possible method to characterise leptospires to serovar level. RAPD-HRM has recently been used to measure intra-serovar convergence between strains of the same serovar as well as inter-serovar divergence between strains of different serovars. The results indicate that intra-serovar heterogeneity and inter-serovar homogeneity may limit the application of RAPD-HRM in routine diagnostics. They also indicate that genetic attenuation of aged, high-passage-number isolates could undermine the use of RAPD-HRM or any other molecular technology. Such genetic attenuation may account for a general decrease seen in titres of rabbit hyperimmune antibodies over time. Before RAPD-HRM can be further advanced as a routine diagnostic tool, strains more representative of the wild-type serovars of a given region need to be identified. Further, RAPD-HRM analysis of reference strains indicates that the routine renewal of reference collections, with new isolates, may be needed to maintain the genetic integrity of the collections.

  9. Rapid identification of tomato Sw-5 resistance-breaking isolates of Tomato spotted wilt virus using high resolution melting and TaqMan SNP Genotyping assays as allelic discrimination techniques.

    Directory of Open Access Journals (Sweden)

    Valentina di Rienzo

    Full Text Available In tomato, resistance to Tomato spotted wilt virus (TSWV is conferred by the dominant gene, designated Sw-5. Virulent Sw-5 resistance breaking (SRB mutants of TSWV have been reported on Sw-5 tomato cultivars. Two different PCR-based allelic discrimination techniques, namely Custom TaqMan™ SNP Genotyping and high-resolution melting (HRM assays, were developed and compared for their ability to distinguish between avirulent (Sw-5 non-infecting, SNI and SRB biotypes. TaqMan assays proved to be more sensitive (threshold of detection in a range of 50-70 TSWV RNA copies and more reliable than HRM, assigning 25 TSWV isolates to their correct genotype with an accuracy of 100%. Moreover, the TaqMan SNP assays were further improved developing a rapid and simple protocol that included crude leaf extraction for RNA template preparations. On the other hand, HRM assays showed higher levels of sensitivity than TaqMan when used to co-detect both biotypes in different artificial mixtures. These diagnostic assays contributed to gain preliminary information on the epidemiology of TSWV isolates in open field conditions. In fact, the presented data suggest that SRB isolates are present as stable populations established year round, persisting on both winter (globe artichoke and summer (tomato crops, in the same cultivated areas of Southern Italy.

  10. High resolution data acquisition

    Science.gov (United States)

    Thornton, Glenn W.; Fuller, Kenneth R.

    1993-01-01

    A high resolution event interval timing system measures short time intervals such as occur in high energy physics or laser ranging. Timing is provided from a clock (38) pulse train (37) and analog circuitry (44) for generating a triangular wave (46) synchronously with the pulse train (37). The triangular wave (46) has an amplitude and slope functionally related to the time elapsed during each clock pulse in the train. A converter (18, 32) forms a first digital value of the amplitude and slope of the triangle wave at the start of the event interval and a second digital value of the amplitude and slope of the triangle wave at the end of the event interval. A counter (26) counts the clock pulse train (37) during the interval to form a gross event interval time. A computer (52) then combines the gross event interval time and the first and second digital values to output a high resolution value for the event interval.

  11. ANL high resolution injector

    International Nuclear Information System (INIS)

    Minehara, E.; Kutschera, W.; Hartog, P.D.; Billquist, P.

    1985-01-01

    The ANL (Argonne National Laboratory) high-resolution injector has been installed to obtain higher mass resolution and higher preacceleration, and to utilize effectively the full mass range of ATLAS (Argonne Tandem Linac Accelerator System). Preliminary results of the first beam test are reported briefly. The design and performance, in particular a high-mass-resolution magnet with aberration compensation, are discussed. 7 refs., 5 figs., 2 tabs

  12. Ultra high resolution tomography

    Energy Technology Data Exchange (ETDEWEB)

    Haddad, W.S.

    1994-11-15

    Recent work and results on ultra high resolution three dimensional imaging with soft x-rays will be presented. This work is aimed at determining microscopic three dimensional structure of biological and material specimens. Three dimensional reconstructed images of a microscopic test object will be presented; the reconstruction has a resolution on the order of 1000 A in all three dimensions. Preliminary work with biological samples will also be shown, and the experimental and numerical methods used will be discussed.

  13. High resolution (transformers.

    Science.gov (United States)

    Garcia-Souto, Jose A; Lamela-Rivera, Horacio

    2006-10-16

    A novel fiber-optic interferometric sensor is presented for vibrations measurements and analysis. In this approach, it is shown applied to the vibrations of electrical structures within power transformers. A main feature of the sensor is that an unambiguous optical phase measurement is performed using the direct detection of the interferometer output, without external modulation, for a more compact and stable implementation. High resolution of the interferometric measurement is obtained with this technique (transformers are also highlighted.

  14. High resolution melting curve analysis targeting the HBB gene mutational hot-spot offers a reliable screening approach for all common as well as most of the rare beta-globin gene mutations in Bangladesh.

    Science.gov (United States)

    Islam, Md Tarikul; Sarkar, Suprovath Kumar; Sultana, Nusrat; Begum, Mst Noorjahan; Bhuyan, Golam Sarower; Talukder, Shezote; Muraduzzaman, A K M; Alauddin, Md; Islam, Mohammad Sazzadul; Biswas, Pritha Promita; Biswas, Aparna; Qadri, Syeda Kashfi; Shirin, Tahmina; Banu, Bilquis; Sadya, Salma; Hussain, Manzoor; Sarwardi, Golam; Khan, Waqar Ahmed; Mannan, Mohammad Abdul; Shekhar, Hossain Uddin; Chowdhury, Emran Kabir; Sajib, Abu Ashfaqur; Akhteruzzaman, Sharif; Qadri, Syed Saleheen; Qadri, Firdausi; Mannoor, Kaiissar

    2018-01-02

    Bangladesh lies in the global thalassemia belt, which has a defined mutational hot-spot in the beta-globin gene. The high carrier frequencies of beta-thalassemia trait and hemoglobin E-trait in Bangladesh necessitate a reliable DNA-based carrier screening approach that could supplement the use of hematological and electrophoretic indices to overcome the barriers of carrier screening. With this view in mind, the study aimed to establish a high resolution melting (HRM) curve-based rapid and reliable mutation screening method targeting the mutational hot-spot of South Asian and Southeast Asian countries that encompasses exon-1 (c.1 - c.92), intron-1 (c.92 + 1 - c.92 + 130) and a portion of exon-2 (c.93 - c.217) of the HBB gene which harbors more than 95% of mutant alleles responsible for beta-thalassemia in Bangladesh. Our HRM approach could successfully differentiate ten beta-globin gene mutations, namely c.79G > A, c.92 + 5G > C, c.126_129delCTTT, c.27_28insG, c.46delT, c.47G > A, c.92G > C, c.92 + 130G > C, c.126delC and c.135delC in heterozygous states from the wild type alleles, implying the significance of the approach for carrier screening as the first three of these mutations account for ~85% of total mutant alleles in Bangladesh. Moreover, different combinations of compound heterozygous mutations were found to generate melt curves that were distinct from the wild type alleles and from one another. Based on the findings, sixteen reference samples were run in parallel to 41 unknown specimens to perform direct genotyping of the beta-thalassemia specimens using HRM. The HRM-based genotyping of the unknown specimens showed 100% consistency with the sequencing result. Targeting the mutational hot-spot, the HRM approach could be successfully applied for screening of beta-thalassemia carriers in Bangladesh as well as in other countries of South Asia and Southeast Asia. The approach could be a useful supplement of hematological and

  15. Quantitative PCR high-resolution melting (qPCR-HRM) curve analysis, a new approach to simultaneously screen point mutations and large rearrangements: application to MLH1 germline mutations in Lynch syndrome.

    Science.gov (United States)

    Rouleau, Etienne; Lefol, Cédrick; Bourdon, Violaine; Coulet, Florence; Noguchi, Tetsuro; Soubrier, Florent; Bièche, Ivan; Olschwang, Sylviane; Sobol, Hagay; Lidereau, Rosette

    2009-06-01

    Several techniques have been developed to screen mismatch repair (MMR) genes for deleterious mutations. Until now, two different techniques were required to screen for both point mutations and large rearrangements. For the first time, we propose a new approach, called "quantitative PCR (qPCR) high-resolution melting (HRM) curve analysis (qPCR-HRM)," which combines qPCR and HRM to obtain a rapid and cost-effective method suitable for testing a large series of samples. We designed PCR amplicons to scan the MLH1 gene using qPCR HRM. Seventy-six patients were fully scanned in replicate, including 14 wild-type patients and 62 patients with known mutations (57 point mutations and five rearrangements). To validate the detected mutations, we used sequencing and/or hybridization on a dedicated MLH1 array-comparative genomic hybridization (array-CGH). All point mutations and rearrangements detected by denaturing high-performance liquid chromatography (dHPLC)+multiplex ligation-dependent probe amplification (MLPA) were successfully detected by qPCR HRM. Three large rearrangements were characterized with the dedicated MLH1 array-CGH. One variant was detected with qPCR HRM in a wild-type patient and was located within the reverse primer. One variant was not detected with qPCR HRM or with dHPLC due to its proximity to a T-stretch. With qPCR HRM, prescreening for point mutations and large rearrangements are performed in one tube and in one step with a single machine, without the need for any automated sequencer in the prescreening process. In replicate, its reagent cost, sensitivity, and specificity are comparable to those of dHPLC+MLPA techniques. However, qPCR HRM outperformed the other techniques in terms of its rapidity and amount of data provided.

  16. Development of melting temperature-based SYBR Green I polymerase chain reaction methods for multiplex genetically modified organism detection.

    Science.gov (United States)

    Hernández, Marta; Rodríguez-Lázaro, David; Esteve, Teresa; Prat, Salomé; Pla, Maria

    2003-12-15

    Commercialization of several genetically modified crops has been approved worldwide to date. Uniplex polymerase chain reaction (PCR)-based methods to identify these different insertion events have been developed, but their use in the analysis of all commercially available genetically modified organisms (GMOs) is becoming progressively insufficient. These methods require a large number of assays to detect all possible GMOs present in the sample and thereby the development of multiplex PCR systems using combined probes and primers targeted to sequences specific to various GMOs is needed for detection of this increasing number of GMOs. Here we report on the development of a multiplex real-time PCR suitable for multiple GMO identification, based on the intercalating dye SYBR Green I and the analysis of the melting curves of the amplified products. Using this method, different amplification products specific for Maximizer 176, Bt11, MON810, and GA21 maize and for GTS 40-3-2 soybean were obtained and identified by their specific Tm. We have combined amplification of these products in a number of multiplex reactions and show the suitability of the methods for identification of GMOs with a sensitivity of 0.1% in duplex reactions. The described methods offer an economic and simple alternative to real-time PCR systems based on sequence-specific probes (i.e., TaqMan chemistry). These methods can be used as selection tests and further optimized for uniplex GMO quantification.

  17. High resolution ultrasonic densitometer

    International Nuclear Information System (INIS)

    Dress, W.B.

    1983-01-01

    The velocity of torsional stress pulses in an ultrasonic waveguide of non-circular cross section is affected by the temperature and density of the surrounding medium. Measurement of the transit times of acoustic echoes from the ends of a sensor section are interpreted as level, density, and temperature of the fluid environment surrounding that section. This paper examines methods of making these measurements to obtain high resolution, temperature-corrected absolute and relative density and level determinations of the fluid. Possible applications include on-line process monitoring, a hand-held density probe for battery charge state indication, and precise inventory control for such diverse fluids as uranium salt solutions in accountability storage and gasoline in service station storage tanks

  18. Analysis of RET promoter CpG island methylation using methylation-specific PCR (MSP), pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM): impact on stage II colon cancer patient outcome.

    Science.gov (United States)

    Draht, Muriel X G; Smits, Kim M; Jooste, Valérie; Tournier, Benjamin; Vervoort, Martijn; Ramaekers, Chantal; Chapusot, Caroline; Weijenberg, Matty P; van Engeland, Manon; Melotte, Veerle

    2016-01-01

    Already since the 1990s, promoter CpG island methylation markers have been considered promising diagnostic, prognostic, and predictive cancer biomarkers. However, so far, only a limited number of DNA methylation markers have been introduced into clinical practice. One reason why the vast majority of methylation markers do not translate into clinical applications is lack of independent validation of methylation markers, often caused by differences in methylation analysis techniques. We recently described RET promoter CpG island methylation as a potential prognostic marker in stage II colorectal cancer (CRC) patients of two independent series. In the current study, we analyzed the RET promoter CpG island methylation of 241 stage II colon cancer patients by direct methylation-specific PCR (MSP), nested-MSP, pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM). All primers were designed as close as possible to the same genomic region. In order to investigate the effect of different DNA methylation assays on patient outcome, we assessed the clinical sensitivity and specificity as well as the association of RET methylation with overall survival for three and five years of follow-up. Using direct-MSP and nested-MSP, 12.0 % (25/209) and 29.6 % (71/240) of the patients showed RET promoter CpG island methylation. Methylation frequencies detected by pyrosequencing were related to the threshold for positivity that defined RET methylation. Methylation frequencies obtained by pyrosequencing (threshold for positivity at 20 %) and MS-HRM were 13.3 % (32/240) and 13.8 % (33/239), respectively. The pyrosequencing threshold for positivity of 20 % showed the best correlation with MS-HRM and direct-MSP results. Nested-MSP detected RET promoter CpG island methylation in deceased patients with a higher sensitivity (33.1 %) compared to direct-MSP (10.7 %), pyrosequencing (14.4 %), and MS-HRM (15.4 %). While RET methylation frequencies detected by nested

  19. Validation of High Resolution Melting Analysis (HRM of the Amplified ITS2 Region for the Detection and Identification of Yeasts from Clinical Samples: Comparison with Culture and MALDI-TOF Based Identification.

    Directory of Open Access Journals (Sweden)

    Hans Duyvejonck

    Full Text Available Candida species are known as opportunistic pathogens, and a possible cause of invasive infections. Because of their species-specific antimycotic resistance patterns, reliable techniques for their detection, quantification and identification are needed. We validated a DNA amplification method for direct detection of Candida spp. from clinical samples, namely the ITS2-High Resolution Melting Analysis (direct method, by comparing it with a culture and MALDI-TOF Mass Spectrometry based method (indirect method to establish the presence of Candida species in three different types of clinical samples.A total of 347 clinical samples, i.e. throat swabs, rectal swabs and vaginal swabs, were collected from the gynaecology/obstetrics, intensive care and haematology wards at the Ghent University Hospital, Belgium. For the direct method, ITS2-HRM was preceded by NucliSENS easyMAG DNA extraction, directly on the clinical samples. For the indirect method, clinical samples were cultured on Candida ID and individual colonies were identified by MALDI-TOF.For 83.9% of the samples there was complete concordance between both techniques, i.e. the same Candida species were detected in 31.1% of the samples or no Candida species were detected in 52.8% of the samples. In 16.1% of the clinical samples, discrepant results were obtained, of which only 6.01% were considered as major discrepancies. Discrepancies occurred mostly when overall numbers of Candida cells in the samples were low and/or when multiple species were present in the sample.Most of the discrepancies could be decided in the advantage of the direct method. This is due to samples in which no yeast could be cultured whereas low amounts could be detected by the direct method and to samples in which high quantities of Candida robusta according to ITS2-HRM were missed by culture on Candida ID agar. It remains to be decided whether the diagnostic advantages of the direct method compensate for its disadvantages.

  20. Miltenberger blood group typing by real-time polymerase chain reaction (qPCR) melting curve analysis in Thai population.

    Science.gov (United States)

    Vongsakulyanon, A; Kitpoka, P; Kunakorn, M; Srikhirin, T

    2015-12-01

    To develop reliable and convenient methods for Miltenberger (Mi(a) ) blood group typing. To apply real-time polymerase chain reaction (qPCR) melting curve analysis to Mi(a) blood group typing. The Mi(a) blood group is the collective set of glycophorin hybrids in the MNS blood group system. Mi(a+) blood is common among East Asians and is also found in the Thai population. Incompatible Mi(a) blood transfusions pose the risk of life-threatening haemolysis; therefore, Mi(a) blood group typing is necessary in ethnicities where the Mi(a) blood group is prevalent. One hundred and forty-three blood samples from Thai blood donors were used in the study. The samples included 50 Mi(a+) samples and 93 Mi(a-) samples, which were defined by serology. The samples were typed by Mi(a) typing qPCR, and 50 Mi(a+) samples were sequenced to identify the Mi(a) subtypes. Mi(a) subtyping qPCR was performed to define GP.Mur. Both Mi(a) typing and Mi(a) subtyping were tested on a conventional PCR platform. The results of Mi(a) typing qPCR were all concordant with serology. Sequencing of the 50 Mi(a+) samples revealed 47 GP.Mur samples and 3 GP.Hop or Bun samples. Mi(a) subtyping qPCR was the supplementary test used to further define GP.Mur from other Mi(a) subtypes. Both Mi(a) typing and Mi(a) subtyping performed well using a conventional PCR platform. Mi(a) typing qPCR correctly identified Mi(a) blood groups in a Thai population with the feasibility of Mi(a) subtype discrimination, and Mi(a) subtyping qPCR was able to further define GP.Mur from other Mi(a) subtypes. © 2015 British Blood Transfusion Society.

  1. Berkeley High-Resolution Ball

    International Nuclear Information System (INIS)

    Diamond, R.M.

    1984-10-01

    Criteria for a high-resolution γ-ray system are discussed. Desirable properties are high resolution, good response function, and moderate solid angle so as to achieve not only double- but triple-coincidences with good statistics. The Berkeley High-Resolution Ball involved the first use of bismuth germanate (BGO) for anti-Compton shield for Ge detectors. The resulting compact shield permitted rather close packing of 21 detectors around a target. In addition, a small central BGO ball gives the total γ-ray energy and multiplicity, as well as the angular pattern of the γ rays. The 21-detector array is nearly complete, and the central ball has been designed, but not yet constructed. First results taken with 9 detector modules are shown for the nucleus 156 Er. The complex decay scheme indicates a transition from collective rotation (prolate shape) to single- particle states (possibly oblate) near spin 30 h, and has other interesting features

  2. High resolution metric imaging payload

    Science.gov (United States)

    Delclaud, Y.

    2017-11-01

    Alcatel Space Industries has become Europe's leader in the field of high and very high resolution optical payloads, in the frame work of earth observation system able to provide military government with metric images from space. This leadership allowed ALCATEL to propose for the export market, within a French collaboration frame, a complete space based system for metric observation.

  3. Real-time polymerase chain reaction with melting analysis of positive blood culture specimens in bloodstream infections: diagnostic value and turnaround time.

    Science.gov (United States)

    Angeletti, Silvia; Gherardi, Giovanni; De Florio, Lucia; Avola, Alessandra; Crea, Francesca; Riva, Elisabetta; Vitali, Massimiliano Andrea; Galluzzo, Sara; Dicuonzo, Giordano

    2013-01-01

    A Real-time polymerase chain reaction (PCR) with melting analysis was devised to target bacterial and fungal genes together with the most prevalent antimicrobial resistance genes in 250 positive blood culture broths. This method allowed the blood culture cultivated pathogens to be classified into clinically relevant groups such as Enterobacteriaceae, oxidase-positive bacilli, oxidase-positive coccobacilli, S. aureus and yeast. Enterococci and streptococci could be distinguished from CoNS only by the Gram stain. Gram-positive bacilli were discriminated from Gram-positive cocci by Gram stain. Furthermore, the most important antimicrobial resistant genes such as mecA, vanA, bla TEM , bla SHV and bla CTX-M could be identified. All results were obtained with a turnaround time of three hours from the moment of blood culture positivity compared to 24-72 hours for phenotypic methods. In conclusion, the proposed approach can allow the clinician to implement proper early management of sepsis patients.

  4. Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis.

    Science.gov (United States)

    Hofmann, Joerg; Frenzel, Katrin; Minh, Bui Q; von Haeseler, Arndt; Edelmann, Anke; Ross, Stefan R; Berg, Thomas; Krüger, Detlev H; Meisel, Helga

    2010-06-01

    Hepatitis D virus (HDV) infection is an important etiologic agent of fulminant hepatitis and may aggravate the clinical course of chronic hepatitis B infection resulting in cirrhosis and liver failure. This report describes the establishment of a real-time reverse transcriptase polymerase chain reaction method that allows the quantitative detection of HDV-1 and HDV-3 with a sensitivity in a linear range of 2 x 10(3) to 10(8) copies/mL. Additionally, the new assay provides the opportunity to distinguish HDV-1 from HDV-3 by a subsequent melting curve analysis, an important option because these HDV types are highly associated with severe clinical outcome. The results of the melting curve analysis of 42 HDV sequences obtained in this study and the phylogenetic analysis based on 139 full-length sequences from GenBank were consistent and showed that all sequences described here cluster within the HDV-1 clade. Therefore, this assay is useful for monitoring of antiviral treatment and molecular epidemiologic studies of HDV distribution. Copyright 2010 Elsevier Inc. All rights reserved.

  5. The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and discrimination of nematodes Bursaphelenchus xylophilus and Bursaphelenchus mucronatus.

    Science.gov (United States)

    Filipiak, Anna; Hasiów-Jaroszewska, Beata

    2016-04-01

    The real-time PCR-HRM analysis was developed for the detection and discrimination of the quarantine nematode Bursaphelenchus xylophilus and Bursaphelenchus mucronatus. A set of primers was designed to target the ITS region of rDNA. The results have demonstrated that this analysis is a valuable tool for differentiation of these both species. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Using surface-enhanced Raman spectroscopy and electrochemically driven melting to discriminate Yersinia pestis from Y. pseudotuberculosis based on single nucleotide polymorphisms within unpurified polymerase chain reaction amplicons.

    Science.gov (United States)

    Papadopoulou, Evanthia; Goodchild, Sarah A; Cleary, David W; Weller, Simon A; Gale, Nittaya; Stubberfield, Michael R; Brown, Tom; Bartlett, Philip N

    2015-02-03

    The development of sensors for the detection of pathogen-specific DNA, including relevant species/strain level discrimination, is critical in molecular diagnostics with major impacts in areas such as bioterrorism and food safety. Herein, we use electrochemically driven denaturation assays monitored by surface-enhanced Raman spectroscopy (SERS) to target single nucleotide polymorphisms (SNPs) that distinguish DNA amplicons generated from Yersinia pestis, the causative agent of plague, from the closely related species Y. pseudotuberculosis. Two assays targeting SNPs within the groEL and metH genes of these two species have been successfully designed. Polymerase chain reaction (PCR) was used to produce Texas Red labeled single-stranded DNA (ssDNA) amplicons of 262 and 251 bases for the groEL and metH targets, respectively. These amplicons were used in an unpurified form to hybridize to immobilized probes then subjected to electrochemically driven melting. In all cases electrochemically driven melting was able to discriminate between fully homologous DNA and that containing SNPs. The metH assay was particularly challenging due to the presence of only a single base mismatch in the middle of the 251 base long PCR amplicon. However, manipulation of assay conditions (conducting the electrochemical experiments at 10 °C) resulted in greater discrimination between the complementary and mismatched DNA. Replicate data were collected and analyzed for each duplex on different days, using different batches of PCR product and different sphere segment void (SSV) substrates. Despite the variability introduced by these differences, the assays are shown to be reliable and robust providing a new platform for strain discrimination using unpurified PCR samples.

  7. High-resolution ultrasonic spectroscopy

    Directory of Open Access Journals (Sweden)

    V. Buckin

    2018-03-01

    Full Text Available High-resolution ultrasonic spectroscopy (HR-US is an analytical technique for direct and non-destructive monitoring of molecular and micro-structural transformations in liquids and semi-solid materials. It is based on precision measurements of ultrasonic velocity and attenuation in analysed samples. The application areas of HR-US in research, product development, and quality and process control include analysis of conformational transitions of polymers, ligand binding, molecular self-assembly and aggregation, crystallisation, gelation, characterisation of phase transitions and phase diagrams, and monitoring of chemical and biochemical reactions. The technique does not require optical markers or optical transparency. The HR-US measurements can be performed in small sample volumes (down to droplet size, over broad temperature range, at ambient and elevated pressures, and in various measuring regimes such as automatic temperature ramps, titrations and measurements in flow.

  8. High Resolution Thermometry for EXACT

    Science.gov (United States)

    Panek, J. S.; Nash, A. E.; Larson, M.; Mulders, N.

    2000-01-01

    High Resolution Thermometers (HRTs) based on SQUID detection of the magnetization of a paramagnetic salt or a metal alloy has been commonly used for sub-nano Kelvin temperature resolution in low temperature physics experiments. The main applications to date have been for temperature ranges near the lambda point of He-4 (2.177 K). These thermometers made use of materials such as Cu(NH4)2Br4 *2H2O, GdCl3, or PdFe. None of these materials are suitable for EXACT, which will explore the region of the He-3/He-4 tricritical point at 0.87 K. The experiment requirements and properties of several candidate paramagnetic materials will be presented, as well as preliminary test results.

  9. High resolution tomographic instrument development

    International Nuclear Information System (INIS)

    1992-01-01

    Our recent work has concentrated on the development of high-resolution PET instrumentation reflecting in part the growing importance of PET in nuclear medicine imaging. We have developed a number of positron imaging instruments and have the distinction that every instrument has been placed in operation and has had an extensive history of application for basic research and clinical study. The present program is a logical continuation of these earlier successes. PCR-I, a single ring positron tomograph was the first demonstration of analog coding using BGO. It employed 4 mm detectors and is currently being used for a wide range of biological studies. These are of immense importance in guiding the direction for future instruments. In particular, PCR-II, a volume sensitive positron tomograph with 3 mm spatial resolution has benefited greatly from the studies using PCR-I. PCR-II is currently in the final stages of assembly and testing and will shortly be placed in operation for imaging phantoms, animals and ultimately humans. Perhaps the most important finding resulting from our previous study is that resolution and sensitivity must be carefully balanced to achieve a practical high resolution system. PCR-II has been designed to have the detection characteristics required to achieve 3 mm resolution in human brain under practical imaging situations. The development of algorithms by the group headed by Dr. Chesler is based on a long history of prior study including his joint work with Drs. Pelc and Reiderer and Stearns. This body of expertise will be applied to the processing of data from PCR-II when it becomes operational

  10. High resolution tomographic instrument development

    Energy Technology Data Exchange (ETDEWEB)

    1992-08-01

    Our recent work has concentrated on the development of high-resolution PET instrumentation reflecting in part the growing importance of PET in nuclear medicine imaging. We have developed a number of positron imaging instruments and have the distinction that every instrument has been placed in operation and has had an extensive history of application for basic research and clinical study. The present program is a logical continuation of these earlier successes. PCR-I, a single ring positron tomograph was the first demonstration of analog coding using BGO. It employed 4 mm detectors and is currently being used for a wide range of biological studies. These are of immense importance in guiding the direction for future instruments. In particular, PCR-II, a volume sensitive positron tomograph with 3 mm spatial resolution has benefited greatly from the studies using PCR-I. PCR-II is currently in the final stages of assembly and testing and will shortly be placed in operation for imaging phantoms, animals and ultimately humans. Perhaps the most important finding resulting from our previous study is that resolution and sensitivity must be carefully balanced to achieve a practical high resolution system. PCR-II has been designed to have the detection characteristics required to achieve 3 mm resolution in human brain under practical imaging situations. The development of algorithms by the group headed by Dr. Chesler is based on a long history of prior study including his joint work with Drs. Pelc and Reiderer and Stearns. This body of expertise will be applied to the processing of data from PCR-II when it becomes operational.

  11. High resolution tomographic instrument development

    Energy Technology Data Exchange (ETDEWEB)

    1992-01-01

    Our recent work has concentrated on the development of high-resolution PET instrumentation reflecting in part the growing importance of PET in nuclear medicine imaging. We have developed a number of positron imaging instruments and have the distinction that every instrument has been placed in operation and has had an extensive history of application for basic research and clinical study. The present program is a logical continuation of these earlier successes. PCR-I, a single ring positron tomograph was the first demonstration of analog coding using BGO. It employed 4 mm detectors and is currently being used for a wide range of biological studies. These are of immense importance in guiding the direction for future instruments. In particular, PCR-II, a volume sensitive positron tomograph with 3 mm spatial resolution has benefited greatly from the studies using PCR-I. PCR-II is currently in the final stages of assembly and testing and will shortly be placed in operation for imaging phantoms, animals and ultimately humans. Perhaps the most important finding resulting from our previous study is that resolution and sensitivity must be carefully balanced to achieve a practical high resolution system. PCR-II has been designed to have the detection characteristics required to achieve 3 mm resolution in human brain under practical imaging situations. The development of algorithms by the group headed by Dr. Chesler is based on a long history of prior study including his joint work with Drs. Pelc and Reiderer and Stearns. This body of expertise will be applied to the processing of data from PCR-II when it becomes operational.

  12. High-Resolution Mass Spectrometers

    Science.gov (United States)

    Marshall, Alan G.; Hendrickson, Christopher L.

    2008-07-01

    Over the past decade, mass spectrometry has been revolutionized by access to instruments of increasingly high mass-resolving power. For small molecules up to ˜400 Da (e.g., drugs, metabolites, and various natural organic mixtures ranging from foods to petroleum), it is possible to determine elemental compositions (CcHhNnOoSsPp…) of thousands of chemical components simultaneously from accurate mass measurements (the same can be done up to 1000 Da if additional information is included). At higher mass, it becomes possible to identify proteins (including posttranslational modifications) from proteolytic peptides, as well as lipids, glycoconjugates, and other biological components. At even higher mass (˜100,000 Da or higher), it is possible to characterize posttranslational modifications of intact proteins and to map the binding surfaces of large biomolecule complexes. Here we review the principles and techniques of the highest-resolution analytical mass spectrometers (time-of-flight and Fourier transform ion cyclotron resonance and orbitrap mass analyzers) and describe some representative high-resolution applications.

  13. High-Resolution Integrated Optical System

    Science.gov (United States)

    Prakapenka, V. B.; Goncharov, A. F.; Holtgrewe, N.; Greenberg, E.

    2017-12-01

    Raman and optical spectroscopy in-situ at extreme high pressure and temperature conditions relevant to the planets' deep interior is a versatile tool for characterization of wide range of properties of minerals essential for understanding the structure, composition, and evolution of terrestrial and giant planets. Optical methods, greatly complementing X-ray diffraction and spectroscopy techniques, become crucial when dealing with light elements. Study of vibrational and optical properties of minerals and volatiles, was a topic of many research efforts in past decades. A great deal of information on the materials properties under extreme pressure and temperature has been acquired including that related to structural phase changes, electronic transitions, and chemical transformations. These provide an important insight into physical and chemical states of planetary interiors (e.g. nature of deep reservoirs) and their dynamics including heat and mass transport (e.g. deep carbon cycle). Optical and vibrational spectroscopy can be also very instrumental for elucidating the nature of the materials molten states such as those related to the Earth's volatiles (CO2, CH4, H2O), aqueous fluids and silicate melts, planetary ices (H2O, CH4, NH3), noble gases, and H2. The optical spectroscopy study performed concomitantly with X-ray diffraction and spectroscopy measurements at the GSECARS beamlines on the same sample and at the same P-T conditions would greatly enhance the quality of this research and, moreover, will provide unique new information on chemical state of matter. The advanced high-resolution user-friendly integrated optical system is currently under construction and expected to be completed by 2018. In our conceptual design we have implemented Raman spectroscopy with five excitation wavelengths (266, 473, 532, 660, 946 nm), confocal imaging, double sided IR laser heating combined with high temperature Raman (including coherent anti-Stokes Raman scattering) and

  14. High-resolution intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Volker Andresen

    Full Text Available Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and

  15. High-Resolution Intravital Microscopy

    Science.gov (United States)

    Andresen, Volker; Pollok, Karolin; Rinnenthal, Jan-Leo; Oehme, Laura; Günther, Robert; Spiecker, Heinrich; Radbruch, Helena; Gerhard, Jenny; Sporbert, Anje; Cseresnyes, Zoltan; Hauser, Anja E.; Niesner, Raluca

    2012-01-01

    Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy - the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology

  16. Section on High Resolution Optical Imaging (HROI)

    Data.gov (United States)

    Federal Laboratory Consortium — The Section on High Resolution Optical Imaging (HROI) develops novel technologies for studying biological processes at unprecedented speed and resolution. Research...

  17. Development of AMS high resolution injector system

    International Nuclear Information System (INIS)

    Bao Yiwen; Guan Xialing; Hu Yueming

    2008-01-01

    The Beijing HI-13 tandem accelerator AMS high resolution injector system was developed. The high resolution energy achromatic system consists of an electrostatic analyzer and a magnetic analyzer, which mass resolution can reach 600 and transmission is better than 80%. (authors)

  18. Comparison of Direct Sequencing, Real-Time PCR-High Resolution Melt (PCR-HRM) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis for Genotyping of Common Thiopurine Intolerant Variant Alleles NUDT15 c.415C>T and TPMT c.719A>G (TPMT*3C).

    Science.gov (United States)

    Fong, Wai-Ying; Ho, Chi-Chun; Poon, Wing-Tat

    2017-05-12

    Thiopurine intolerance and treatment-related toxicity, such as fatal myelosuppression, is related to non-function genetic variants encoding thiopurine S-methyltransferase (TPMT) and Nudix hydrolase 15 (NUDT15). Genetic testing of the common variants NUDT15:NM_018283.2:c.415C>T (Arg139Cys, dbSNP rs116855232 T allele) and TPMT: NM_000367.4:c.719A>G (TPMT*3C, dbSNP rs1142345 G allele) in East Asians including Chinese can potentially prevent treatment-related complications. Two complementary genotyping approaches, real-time PCR-high resolution melt (PCR-HRM) and PCR-restriction fragment length morphism (PCR-RFLP) analysis were evaluated using conventional PCR and Sanger sequencing genotyping as the gold standard. Sixty patient samples were tested, revealing seven patients (11.7%) heterozygous for NUDT15 c.415C>T, one patient homozygous for the variant and one patient heterozygous for the TPMT*3C non-function allele. No patient was found to harbor both variants. In total, nine out of 60 (15%) patients tested had genotypic evidence of thiopurine intolerance, which may require dosage adjustment or alternative medication should they be started on azathioprine, mercaptopurine or thioguanine. The two newly developed assays were more efficient and showed complete concordance (60/60, 100%) compared to the Sanger sequencing results. Accurate and cost-effective genotyping assays by real-time PCR-HRM and PCR-RFLP for NUDT15 c.415C>T and TPMT*3C were successfully developed. Further studies may establish their roles in genotype-informed clinical decision-making in the prevention of morbidity and mortality due to thiopurine intolerance.

  19. High resolution sequence stratigraphy in China

    International Nuclear Information System (INIS)

    Zhang Shangfeng; Zhang Changmin; Yin Yanshi; Yin Taiju

    2008-01-01

    Since high resolution sequence stratigraphy was introduced into China by DENG Hong-wen in 1995, it has been experienced two development stages in China which are the beginning stage of theory research and development of theory research and application, and the stage of theoretical maturity and widely application that is going into. It is proved by practices that high resolution sequence stratigraphy plays more and more important roles in the exploration and development of oil and gas in Chinese continental oil-bearing basin and the research field spreads to the exploration of coal mine, uranium mine and other strata deposits. However, the theory of high resolution sequence stratigraphy still has some shortages, it should be improved in many aspects. The authors point out that high resolution sequence stratigraphy should be characterized quantitatively and modelized by computer techniques. (authors)

  20. High resolution CT of the chest

    Energy Technology Data Exchange (ETDEWEB)

    Barneveld Binkhuysen, F H [Eemland Hospital (Netherlands), Dept. of Radiology

    1996-12-31

    Compared to conventional CT high resolution CT (HRCT) shows several extra anatomical structures which might effect both diagnosis and therapy. The extra anatomical structures were discussed briefly in this article. (18 refs.).

  1. High-resolution spectrometer at PEP

    International Nuclear Information System (INIS)

    Weiss, J.M.; HRS Collaboration.

    1982-01-01

    A description is presented of the High Resolution Spectrometer experiment (PEP-12) now running at PEP. The advanced capabilities of the detector are demonstrated with first physics results expected in the coming months

  2. Structure of high-resolution NMR spectra

    CERN Document Server

    Corio, PL

    2012-01-01

    Structure of High-Resolution NMR Spectra provides the principles, theories, and mathematical and physical concepts of high-resolution nuclear magnetic resonance spectra.The book presents the elementary theory of magnetic resonance; the quantum mechanical theory of angular momentum; the general theory of steady state spectra; and multiple quantum transitions, double resonance and spin echo experiments.Physicists, chemists, and researchers will find the book a valuable reference text.

  3. High-resolution structure of the native histone octamer

    International Nuclear Information System (INIS)

    Wood, Christopher M.; Nicholson, James M.; Lambert, Stanley J.; Chantalat, Laurent; Reynolds, Colin D.; Baldwin, John P.

    2005-01-01

    The high-resolution (1.90 Å) model of the native histone octamer allows structural comparisons to be made with the nucleosome-core particle, along with an identification of a likely core-histone binding site. Crystals of native histone octamers (H2A–H2B)–(H4–H3)–(H3′–H4′)–(H2B′–H2A′) from chick erythrocytes in 2 M KCl, 1.35 M potassium phosphate pH 6.9 diffract X-rays to 1.90 Å resolution, yielding a structure with an R work value of 18.7% and an R free of 22.2%. The crystal space group is P6 5 , the asymmetric unit of which contains one complete octamer. This high-resolution model of the histone-core octamer allows further insight into intermolecular interactions, including water molecules, that dock the histone dimers to the tetramer in the nucleosome-core particle and have relevance to nucleosome remodelling. The three key areas analysed are the H2A′–H3–H4 molecular cluster (also H2A–H3′–H4′), the H4–H2B′ interaction (also H4′–H2B) and the H2A′–H4 β-sheet interaction (also H2A–H4′). The latter of these three regions is important to nucleosome remodelling by RNA polymerase II, as it is shown to be a likely core-histone binding site, and its disruption creates an instability in the nucleosome-core particle. A majority of the water molecules in the high-resolution octamer have positions that correlate to similar positions in the high-resolution nucleosome-core particle structure, suggesting that the high-resolution octamer model can be used for comparative studies with the high-resolution nucleosome-core particle

  4. A high resolution portable spectroscopy system

    International Nuclear Information System (INIS)

    Kulkarni, C.P.; Vaidya, P.P.; Paulson, M.; Bhatnagar, P.V.; Pande, S.S.; Padmini, S.

    2003-01-01

    Full text: This paper describes the system details of a High Resolution Portable Spectroscopy System (HRPSS) developed at Electronics Division, BARC. The system can be used for laboratory class, high-resolution nuclear spectroscopy applications. The HRPSS consists of a specially designed compact NIM bin, with built-in power supplies, accommodating a low power, high resolution MCA, and on-board embedded computer for spectrum building and communication. A NIM based spectroscopy amplifier and a HV module for detector bias are integrated (plug-in) in the bin. The system communicates with a host PC via a serial link. Along-with a laptop PC, and a portable HP-Ge detector, the HRPSS offers a laboratory class performance for portable applications

  5. High-resolution multi-slice PET

    International Nuclear Information System (INIS)

    Yasillo, N.J.; Chintu Chen; Ordonez, C.E.; Kapp, O.H.; Sosnowski, J.; Beck, R.N.

    1992-01-01

    This report evaluates the progress to test the feasibility and to initiate the design of a high resolution multi-slice PET system. The following specific areas were evaluated: detector development and testing; electronics configuration and design; mechanical design; and system simulation. The design and construction of a multiple-slice, high-resolution positron tomograph will provide substantial improvements in the accuracy and reproducibility of measurements of the distribution of activity concentrations in the brain. The range of functional brain research and our understanding of local brain function will be greatly extended when the development of this instrumentation is completed

  6. High resolution Neutron and Synchrotron Powder Diffraction

    International Nuclear Information System (INIS)

    Hewat, A.W.

    1986-01-01

    The use of high-resolution powder diffraction has grown rapidly in the past years, with the development of Rietveld (1967) methods of data analysis and new high-resolution diffractometers and multidetectors. The number of publications in this area has increased from a handful per year until 1973 to 150 per year in 1984, with a ten-year total of over 1000. These papers cover a wide area of solid state-chemistry, physics and materials science, and have been grouped under 20 subject headings, ranging from catalysts to zeolites, and from battery electrode materials to pre-stressed superconducting wires. In 1985 two new high-resolution diffractometers are being commissioned, one at the SNS laboratory near Oxford, and one at the ILL in Grenoble. In different ways these machines represent perhaps the ultimate that can be achieved with neutrons and will permit refinement of complex structures with about 250 parameters and unit cell volumes of about 2500 Angstrom/sp3/. The new European Synchotron Facility will complement the Grenoble neutron diffractometers, and extend the role of high-resolution powder diffraction to the direct solution of crystal structures, pioneered in Sweden

  7. High resolution CT in diffuse lung disease

    International Nuclear Information System (INIS)

    Webb, W.R.

    1995-01-01

    High resolution CT (computerized tomography) was discussed in detail. The conclusions were HRCT is able to define lung anatomy at the secondary lobular level and define a variety of abnormalities in patients with diffuse lung diseases. Evidence from numerous studies indicates that HRCT can play a major role in the assessment of diffuse infiltrative lung disease and is indicate clinically (95 refs.)

  8. Classification of high resolution satellite images

    OpenAIRE

    Karlsson, Anders

    2003-01-01

    In this thesis the Support Vector Machine (SVM)is applied on classification of high resolution satellite images. Sveral different measures for classification, including texture mesasures, 1st order statistics, and simple contextual information were evaluated. Additionnally, the image was segmented, using an enhanced watershed method, in order to improve the classification accuracy.

  9. High resolution CT in diffuse lung disease

    Energy Technology Data Exchange (ETDEWEB)

    Webb, W R [California Univ., San Francisco, CA (United States). Dept. of Radiology

    1996-12-31

    High resolution CT (computerized tomography) was discussed in detail. The conclusions were HRCT is able to define lung anatomy at the secondary lobular level and define a variety of abnormalities in patients with diffuse lung diseases. Evidence from numerous studies indicates that HRCT can play a major role in the assessment of diffuse infiltrative lung disease and is indicate clinically (95 refs.).

  10. High-resolution clean-sc

    NARCIS (Netherlands)

    Sijtsma, P.; Snellen, M.

    2016-01-01

    In this paper a high-resolution extension of CLEAN-SC is proposed: HR-CLEAN-SC. Where CLEAN-SC uses peak sources in “dirty maps” to define so-called source components, HR-CLEAN-SC takes advantage of the fact that source components can likewise be derived from points at some distance from the peak,

  11. A High-Resolution Stopwatch for Cents

    Science.gov (United States)

    Gingl, Z.; Kopasz, K.

    2011-01-01

    A very low-cost, easy-to-make stopwatch is presented to support various experiments in mechanics. The high-resolution stopwatch is based on two photodetectors connected directly to the microphone input of a sound card. Dedicated free open-source software has been developed and made available to download. The efficiency is demonstrated by a free…

  12. Planning for shallow high resolution seismic surveys

    CSIR Research Space (South Africa)

    Fourie, CJS

    2008-11-01

    Full Text Available of the input wave. This information can be used in conjunction with this spreadsheet to aid the geophysicist in designing shallow high resolution seismic surveys to achieve maximum resolution and penetration. This Excel spreadsheet is available free from...

  13. Smartphone microendoscopy for high resolution fluorescence imaging

    Directory of Open Access Journals (Sweden)

    Xiangqian Hong

    2016-09-01

    Full Text Available High resolution optical endoscopes are increasingly used in diagnosis of various medical conditions of internal organs, such as the cervix and gastrointestinal (GI tracts, but they are too expensive for use in resource-poor settings. On the other hand, smartphones with high resolution cameras and Internet access have become more affordable, enabling them to diffuse into most rural areas and developing countries in the past decade. In this paper, we describe a smartphone microendoscope that can take fluorescence images with a spatial resolution of 3.1 μm. Images collected from ex vivo, in vitro and in vivo samples using the device are also presented. The compact and cost-effective smartphone microendoscope may be envisaged as a powerful tool for detecting pre-cancerous lesions of internal organs in low and middle-income countries (LMICs.

  14. High resolution NMR theory and chemical applications

    CERN Document Server

    Becker, Edwin D

    2012-01-01

    High Resolution NMR: Theory and Chemical Applications discusses the principles and theory of nuclear magnetic resonance and how this concept is used in the chemical sciences. This book is written at an intermediate level, with mathematics used to augment verbal descriptions of the phenomena. This text pays attention to developing and interrelating four approaches - the steady state energy levels, the rotating vector picture, the density matrix, and the product operator formalism. The style of this book is based on the assumption that the reader has an acquaintance with the general principles of quantum mechanics, but no extensive background in quantum theory or proficiency in mathematics is required. This book begins with a description of the basic physics, together with a brief account of the historical development of the field. It looks at the study of NMR in liquids, including high resolution NMR in the solid state and the principles of NMR imaging and localized spectroscopy. This book is intended to assis...

  15. High resolution NMR theory and chemical applications

    CERN Document Server

    Becker, Edwin D

    1999-01-01

    High Resolution NMR provides a broad treatment of the principles and theory of nuclear magnetic resonance (NMR) as it is used in the chemical sciences. It is written at an "intermediate" level, with mathematics used to augment, rather than replace, clear verbal descriptions of the phenomena. The book is intended to allow a graduate student, advanced undergraduate, or researcher to understand NMR at a fundamental level, and to see illustrations of the applications of NMR to the determination of the structure of small organic molecules and macromolecules, including proteins. Emphasis is on the study of NMR in liquids, but the treatment also includes high resolution NMR in the solid state and the principles of NMR imaging and localized spectroscopy. Careful attention is given to developing and interrelating four approaches - steady state energy levels, the rotating vector picture, the density matrix, and the product operator formalism. The presentation is based on the assumption that the reader has an acquaintan...

  16. High resolution imaging of boron carbide microstructures

    International Nuclear Information System (INIS)

    MacKinnon, I.D.R.; Aselage, T.; Van Deusen, S.B.

    1986-01-01

    Two samples of boron carbide have been examined using high resolution transmission electron microscopy (HRTEM). A hot-pressed B 13 C 2 sample shows a high density of variable width twins normal to (10*1). Subtle shifts or offsets of lattice fringes along the twin plane and normal to approx.(10*5) were also observed. A B 4 C powder showed little evidence of stacking disorder in crystalline regions

  17. High-Resolution MRI in Rectal Cancer

    International Nuclear Information System (INIS)

    Dieguez, Adriana

    2010-01-01

    High-resolution MRI is the best method of assessing the relation of the rectal tumor with the potential circumferential resection margin (CRM). Therefore it is currently considered the method of choice for local staging of rectal cancer. The primary surgery of rectal cancer is total mesorectal excision (TME), which plane of dissection is formed by the mesorectal fascia surrounding mesorectal fat and rectum. This fascia will determine the circumferential margin of resection. At the same time, high resolution MRI allows adequate pre-operative identification of important prognostic risk factors, improving the selection and indication of therapy for each patient. This information includes, besides the circumferential margin of resection, tumor and lymph node staging, extramural vascular invasion and the description of lower rectal tumors. All these should be described in detail in the report, being part of the discussion in the multidisciplinary team, the place where the decisions involving the patient with rectal cancer will take place. The aim of this study is to provide the information necessary to understand the use of high resolution MRI in the identification of prognostic risk factors in rectal cancer. The technical requirements and standardized report for this study will be describe, as well as the anatomical landmarks of importance for the total mesorectal excision (TME), as we have said is the surgery of choice for rectal cancer. (authors) [es

  18. Ultra-high resolution protein crystallography

    International Nuclear Information System (INIS)

    Takeda, Kazuki; Hirano, Yu; Miki, Kunio

    2010-01-01

    Many protein structures have been determined by X-ray crystallography and deposited with the Protein Data Bank. However, these structures at usual resolution (1.5< d<3.0 A) are insufficient in their precision and quantity for elucidating the molecular mechanism of protein functions directly from structural information. Several studies at ultra-high resolution (d<0.8 A) have been performed with synchrotron radiation in the last decade. The highest resolution of the protein crystals was achieved at 0.54 A resolution for a small protein, crambin. In such high resolution crystals, almost all of hydrogen atoms of proteins and some hydrogen atoms of bound water molecules are experimentally observed. In addition, outer-shell electrons of proteins can be analyzed by the multipole refinement procedure. However, the influence of X-rays should be precisely estimated in order to derive meaningful information from the crystallographic results. In this review, we summarize refinement procedures, current status and perspectives for ultra high resolution protein crystallography. (author)

  19. Detection of Babesia canis vogeli and Hepatozoon canis in canine blood by a single-tube real-time fluorescence resonance energy transfer polymerase chain reaction assay and melting curve analysis.

    Science.gov (United States)

    Kongklieng, Amornmas; Intapan, Pewpan M; Boonmars, Thidarut; Thanchomnang, Tongjit; Janwan, Penchom; Sanpool, Oranuch; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2015-03-01

    A real-time fluorescence resonance energy transfer polymerase chain reaction (qFRET PCR) coupled with melting curve analysis was developed for detection of Babesia canis vogeli and Hepatozoon canis infections in canine blood samples in a single tube assay. The target of the assay was a region within the 18S ribosomal RNA gene amplified in either species by a single pair of primers. Following amplification from the DNA of infected dog blood, a fluorescence melting curve analysis was done. The 2 species, B. canis vogeli and H. canis, could be detected and differentiated in infected dog blood samples (n = 37) with high sensitivity (100%). The detection limit for B. canis vogeli was 15 copies of a positive control plasmid, and for H. canis, it was 150 copies of a positive control plasmid. The assay could simultaneously distinguish the DNA of both parasites from the DNA of controls. Blood samples from 5 noninfected dogs were negative, indicating high specificity. Several samples can be run at the same time. The assay can reduce misdiagnosis and the time associated with microscopic examination, and is not prone to the carryover contamination associated with the agarose gel electrophoresis step of conventional PCR. In addition, this qFRET PCR method would be useful to accurately determine the range of endemic areas or to discover those areas where the 2 parasites co-circulate. © 2015 The Author(s).

  20. High resolution, high speed ultrahigh vacuum microscopy

    International Nuclear Information System (INIS)

    Poppa, Helmut

    2004-01-01

    The history and future of transmission electron microscopy (TEM) is discussed as it refers to the eventual development of instruments and techniques applicable to the real time in situ investigation of surface processes with high resolution. To reach this objective, it was necessary to transform conventional high resolution instruments so that an ultrahigh vacuum (UHV) environment at the sample site was created, that access to the sample by various in situ sample modification procedures was provided, and that in situ sample exchanges with other integrated surface analytical systems became possible. Furthermore, high resolution image acquisition systems had to be developed to take advantage of the high speed imaging capabilities of projection imaging microscopes. These changes to conventional electron microscopy and its uses were slowly realized in a few international laboratories over a period of almost 40 years by a relatively small number of researchers crucially interested in advancing the state of the art of electron microscopy and its applications to diverse areas of interest; often concentrating on the nucleation, growth, and properties of thin films on well defined material surfaces. A part of this review is dedicated to the recognition of the major contributions to surface and thin film science by these pioneers. Finally, some of the important current developments in aberration corrected electron optics and eventual adaptations to in situ UHV microscopy are discussed. As a result of all the path breaking developments that have led to today's highly sophisticated UHV-TEM systems, integrated fundamental studies are now possible that combine many traditional surface science approaches. Combined investigations to date have involved in situ and ex situ surface microscopies such as scanning tunneling microscopy/atomic force microscopy, scanning Auger microscopy, and photoemission electron microscopy, and area-integrating techniques such as x-ray photoelectron

  1. USGS High Resolution Orthoimagery Collection - Historical - National Geospatial Data Asset (NGDA) High Resolution Orthoimagery

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — USGS high resolution orthorectified images from The National Map combine the image characteristics of an aerial photograph with the geometric qualities of a map. An...

  2. Thermodynamics of Oligonucleotide Duplex Melting

    Science.gov (United States)

    Schreiber-Gosche, Sherrie; Edwards, Robert A.

    2009-01-01

    Melting temperatures of oligonucleotides are useful for a number of molecular biology applications, such as the polymerase chain reaction (PCR). Although melting temperatures are often calculated with simplistic empirical equations, application of thermodynamics provides more accurate melting temperatures and an opportunity for students to apply…

  3. High resolution extremity CT for biomechanics modeling

    International Nuclear Information System (INIS)

    Ashby, A.E.; Brand, H.; Hollerbach, K.; Logan, C.M.; Martz, H.E.

    1995-01-01

    With the advent of ever more powerful computing and finite element analysis (FEA) capabilities, the bone and joint geometry detail available from either commercial surface definitions or from medical CT scans is inadequate. For dynamic FEA modeling of joints, precise articular contours are necessary to get appropriate contact definition. In this project, a fresh cadaver extremity was suspended in parafin in a lucite cylinder and then scanned with an industrial CT system to generate a high resolution data set for use in biomechanics modeling

  4. Ultra-high resolution coded wavefront sensor

    KAUST Repository

    Wang, Congli

    2017-06-08

    Wavefront sensors and more general phase retrieval methods have recently attracted a lot of attention in a host of application domains, ranging from astronomy to scientific imaging and microscopy. In this paper, we introduce a new class of sensor, the Coded Wavefront Sensor, which provides high spatio-temporal resolution using a simple masked sensor under white light illumination. Specifically, we demonstrate megapixel spatial resolution and phase accuracy better than 0.1 wavelengths at reconstruction rates of 50 Hz or more, thus opening up many new applications from high-resolution adaptive optics to real-time phase retrieval in microscopy.

  5. High resolution extremity CT for biomechanics modeling

    Energy Technology Data Exchange (ETDEWEB)

    Ashby, A.E.; Brand, H.; Hollerbach, K.; Logan, C.M.; Martz, H.E.

    1995-09-23

    With the advent of ever more powerful computing and finite element analysis (FEA) capabilities, the bone and joint geometry detail available from either commercial surface definitions or from medical CT scans is inadequate. For dynamic FEA modeling of joints, precise articular contours are necessary to get appropriate contact definition. In this project, a fresh cadaver extremity was suspended in parafin in a lucite cylinder and then scanned with an industrial CT system to generate a high resolution data set for use in biomechanics modeling.

  6. High-resolution computer-aided moire

    Science.gov (United States)

    Sciammarella, Cesar A.; Bhat, Gopalakrishna K.

    1991-12-01

    This paper presents a high resolution computer assisted moire technique for the measurement of displacements and strains at the microscopic level. The detection of micro-displacements using a moire grid and the problem associated with the recovery of displacement field from the sampled values of the grid intensity are discussed. A two dimensional Fourier transform method for the extraction of displacements from the image of the moire grid is outlined. An example of application of the technique to the measurement of strains and stresses in the vicinity of the crack tip in a compact tension specimen is given.

  7. SPIRAL2/DESIR high resolution mass separator

    Energy Technology Data Exchange (ETDEWEB)

    Kurtukian-Nieto, T., E-mail: kurtukia@cenbg.in2p3.fr [Centre d’Études Nucléaires de Bordeaux Gradignan, Université Bordeaux 1-CNRS/IN2P3, BP 120, F-33175 Gradignan Cedex (France); Baartman, R. [TRIUMF, 4004 Wesbrook Mall, Vancouver B.C., V6T 2A3 (Canada); Blank, B.; Chiron, T. [Centre d’Études Nucléaires de Bordeaux Gradignan, Université Bordeaux 1-CNRS/IN2P3, BP 120, F-33175 Gradignan Cedex (France); Davids, C. [Physics Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Delalee, F. [Centre d’Études Nucléaires de Bordeaux Gradignan, Université Bordeaux 1-CNRS/IN2P3, BP 120, F-33175 Gradignan Cedex (France); Duval, M. [GANIL, CEA/DSM-CNRS/IN2P3, Bd Henri Becquerel, BP 55027, F-14076 Caen Cedex 5 (France); El Abbeir, S.; Fournier, A. [Centre d’Études Nucléaires de Bordeaux Gradignan, Université Bordeaux 1-CNRS/IN2P3, BP 120, F-33175 Gradignan Cedex (France); Lunney, D. [CSNSM-IN2P3-CNRS, Université de Paris Sud, F-91405 Orsay (France); Méot, F. [BNL, Upton, Long Island, New York (United States); Serani, L. [Centre d’Études Nucléaires de Bordeaux Gradignan, Université Bordeaux 1-CNRS/IN2P3, BP 120, F-33175 Gradignan Cedex (France); Stodel, M.-H.; Varenne, F. [GANIL, CEA/DSM-CNRS/IN2P3, Bd Henri Becquerel, BP 55027, F-14076 Caen Cedex 5 (France); and others

    2013-12-15

    DESIR is the low-energy part of the SPIRAL2 ISOL facility under construction at GANIL. DESIR includes a high-resolution mass separator (HRS) with a designed resolving power m/Δm of 31,000 for a 1 π-mm-mrad beam emittance, obtained using a high-intensity beam cooling device. The proposed design consists of two 90-degree magnetic dipoles, complemented by electrostatic quadrupoles, sextupoles, and a multipole, arranged in a symmetric configuration to minimize aberrations. A detailed description of the design and results of extensive simulations are given.

  8. Laboratory of High resolution gamma spectrometry

    International Nuclear Information System (INIS)

    Mendez G, A.; Giber F, J.; Rivas C, I.; Reyes A, B.

    1992-01-01

    The Department of Nuclear Experimentation of the Nuclear Systems Management requests the collaboration of the Engineering unit for the supervision of the execution of the work of the High resolution Gamma spectrometry and low bottom laboratory, using the hut of the sub critic reactor of the Nuclear Center of Mexico. This laboratory has the purpose of determining the activity of special materials irradiated in nuclear power plants. In this report the architecture development, concepts, materials and diagrams for the realization of this type of work are presented. (Author)

  9. High resolution neutron spectroscopy for helium isotopes

    International Nuclear Information System (INIS)

    Abdel-Wahab, M.S.; Klages, H.O.; Schmalz, G.; Haesner, B.H.; Kecskemeti, J.; Schwarz, P.; Wilczynski, J.

    1992-01-01

    A high resolution fast neutron time-of-flight spectrometer is described, neutron time-of-flight spectra are taken using a specially designed TDC in connection to an on-line computer. The high time-of-flight resolution of 5 ps/m enabled the study of the total cross section of 4 He for neutrons near the 3/2 + resonance in the 5 He nucleus. The resonance parameters were determined by a single level Breit-Winger fit to the data. (orig.)

  10. Limiting liability via high resolution image processing

    Energy Technology Data Exchange (ETDEWEB)

    Greenwade, L.E.; Overlin, T.K.

    1996-12-31

    The utilization of high resolution image processing allows forensic analysts and visualization scientists to assist detectives by enhancing field photographs, and by providing the tools and training to increase the quality and usability of field photos. Through the use of digitized photographs and computerized enhancement software, field evidence can be obtained and processed as `evidence ready`, even in poor lighting and shadowed conditions or darkened rooms. These images, which are most often unusable when taken with standard camera equipment, can be shot in the worst of photographic condition and be processed as usable evidence. Visualization scientists have taken the use of digital photographic image processing and moved the process of crime scene photos into the technology age. The use of high resolution technology will assist law enforcement in making better use of crime scene photography and positive identification of prints. Valuable court room and investigation time can be saved and better served by this accurate, performance based process. Inconclusive evidence does not lead to convictions. Enhancement of the photographic capability helps solve one major problem with crime scene photos, that if taken with standard equipment and without the benefit of enhancement software would be inconclusive, thus allowing guilty parties to be set free due to lack of evidence.

  11. High-Resolution Scintimammography: A Pilot Study

    Energy Technology Data Exchange (ETDEWEB)

    Rachel F. Brem; Joelle M. Schoonjans; Douglas A. Kieper; Stan Majewski; Steven Goodman; Cahid Civelek

    2002-07-01

    This study evaluated a novel high-resolution breast-specific gamma camera (HRBGC) for the detection of suggestive breast lesions. Methods: Fifty patients (with 58 breast lesions) for whom a scintimammogram was clinically indicated were prospectively evaluated with a general-purpose gamma camera and a novel HRBGC prototype. The results of conventional and high-resolution nuclear studies were prospectively classified as negative (normal or benign) or positive (suggestive or malignant) by 2 radiologists who were unaware of the mammographic and histologic results. All of the included lesions were confirmed by pathology. Results: There were 30 benign and 28 malignant lesions. The sensitivity for detection of breast cancer was 64.3% (18/28) with the conventional camera and 78.6% (22/28) with the HRBGC. The specificity with both systems was 93.3% (28/30). For the 18 nonpalpable lesions, sensitivity was 55.5% (10/18) and 72.2% (13/18) with the general-purpose camera and the HRBGC, respectively. For lesions 1 cm, 7 of 15 were detected with the general-purpose camera and 10 of 15 with the HRBGC. Four lesions (median size, 8.5 mm) were detected only with the HRBGC and were missed by the conventional camera. Conclusion: Evaluation of indeterminate breast lesions with an HRBGC results in improved sensitivity for the detection of cancer, with greater improvement shown for nonpalpable and 1-cm lesions.

  12. High resolution studies of barium Rydberg states

    International Nuclear Information System (INIS)

    Eliel, E.R.

    1982-01-01

    The subtle structure of Rydberg states of barium with orbital angular momentum 0, 1, 2 and 3 is investigated. Some aspects of atomic theory for a configuration with two valence electrons are reviewed. The Multi Channel Quantum Defect Theory (MQDT) is concisely introduced as a convenient way to describe interactions between Rydberg series. Three high-resolution UV studies are presented. The first two, presenting results on a transition in indium and europium serve as an illustration of the frequency doubling technique. The third study is of hyperfine structure and isotope shifts in low-lying p states in Sr and Ba. An extensive study of the 6snp and 6snf Rydberg states of barium is presented with particular emphasis on the 6snf states. It is shown that the level structure cannot be fully explained with the model introduced earlier. Rather an effective two-body spin-orbit interaction has to be introduced to account for the observed splittings, illustrating that high resolution studies on Rydberg states offer an unique opportunity to determine the importance of such effects. Finally, the 6sns and 6snd series are considered. The hyperfine induced isotope shift in the simple excitation spectra to 6sns 1 S 0 is discussed and attention is paid to series perturbers. It is shown that level mixing parameters can easily be extracted from the experimental data. (Auth.)

  13. Principles of high resolution NMR in solids

    CERN Document Server

    Mehring, Michael

    1983-01-01

    The field of Nuclear Magnetic Resonance (NMR) has developed at a fascinating pace during the last decade. It always has been an extremely valuable tool to the organic chemist by supplying molecular "finger print" spectra at the atomic level. Unfortunately the high resolution achievable in liquid solutions could not be obtained in solids and physicists and physical chemists had to live with unresolved lines open to a wealth of curve fitting procedures and a vast amount of speculations. High resolution NMR in solids seemed to be a paradoxon. Broad structure­ less lines are usually encountered when dealing with NMR in solids. Only with the recent advent of mUltiple pulse, magic angle, cross-polarization, two-dimen­ sional and multiple-quantum spectroscopy and other techniques during the last decade it became possible to resolve finer details of nuclear spin interactions in solids. I have felt that graduate students, researchers and others beginning to get involved with these techniques needed a book which trea...

  14. High-Resolution PET Detector. Final report

    International Nuclear Information System (INIS)

    Karp, Joel

    2014-01-01

    The objective of this project was to develop an understanding of the limits of performance for a high resolution PET detector using an approach based on continuous scintillation crystals rather than pixelated crystals. The overall goal was to design a high-resolution detector, which requires both high spatial resolution and high sensitivity for 511 keV gammas. Continuous scintillation detectors (Anger cameras) have been used extensively for both single-photon and PET scanners, however, these instruments were based on NaI(Tl) scintillators using relatively large, individual photo-multipliers. In this project we investigated the potential of this type of detector technology to achieve higher spatial resolution through the use of improved scintillator materials and photo-sensors, and modification of the detector surface to optimize the light response function.We achieved an average spatial resolution of 3-mm for a 25-mm thick, LYSO continuous detector using a maximum likelihood position algorithm and shallow slots cut into the entrance surface

  15. High Resolution Powder Diffraction and Structure Determination

    International Nuclear Information System (INIS)

    Cox, D. E.

    1999-01-01

    It is clear that high-resolution synchrotrons X-ray powder diffraction is a very powerful and convenient tool for material characterization and structure determination. Most investigations to date have been carried out under ambient conditions and have focused on structure solution and refinement. The application of high-resolution techniques to increasingly complex structures will certainly represent an important part of future studies, and it has been seen how ab initio solution of structures with perhaps 100 atoms in the asymmetric unit is within the realms of possibility. However, the ease with which temperature-dependence measurements can be made combined with improvements in the technology of position-sensitive detectors will undoubtedly stimulate precise in situ structural studies of phase transitions and related phenomena. One challenge in this area will be to develop high-resolution techniques for ultra-high pressure investigations in diamond anvil cells. This will require highly focused beams and very precise collimation in front of the cell down to dimensions of 50 (micro)m or less. Anomalous scattering offers many interesting possibilities as well. As a means of enhancing scattering contrast it has applications not only to the determination of cation distribution in mixed systems such as the superconducting oxides discussed in Section 9.5.3, but also to the location of specific cations in partially occupied sites, such as the extra-framework positions in zeolites, for example. Another possible application is to provide phasing information for ab initio structure solution. Finally, the precise determination of f as a function of energy through an absorption edge can provide useful information about cation oxidation states, particularly in conjunction with XANES data. In contrast to many experiments at a synchrotron facility, powder diffraction is a relatively simple and user-friendly technique, and most of the procedures and software for data analysis

  16. High resolution CT of the lung

    Energy Technology Data Exchange (ETDEWEB)

    Itoh, Harumi (Kyoto Univ. (Japan). Faculty of Medicine)

    1991-02-01

    The emergence of computed tomography (CT) in the early 1970s has greatly contributed to diagnostic radiology. The brain was the first organ examined with CT, followed by the abdomen. For the chest, CT has also come into use shortly after the introduction in the examination of the thoracic cavity and mediastinum. CT techniques were, however, of limited significance in the evaluation of pulmonary diseases, especially diffuse pulmonary diseases. High-resolution CT (HRCT) has been introduced in clinical investigations of the lung field. This article is designed to present chest radiographic and conventional tomographic interpretations and to introduce findings of HRCT corresponding to the same shadows, with a summation of the significance of HRCT and issues of diagnostic imaging. Materials outlined are tuberculosis, pneumoconiosis, bronchopneumonia, mycoplasma pneumonia, lymphangitic carcinomatosis, sarcoidosis, diffuse panbronchiolitis, interstitial pneumonia, and pulmonary emphysema. Finally, an overview of basic investigations evolved from HRCT is given. (N.K.) 140 refs.

  17. Constructing a WISE High Resolution Galaxy Atlas

    Science.gov (United States)

    Jarrett, T. H.; Masci, F.; Tsai, C. W.; Petty, S.; Cluver, M.; Assef, Roberto J.; Benford, D.; Blain, A.; Bridge, C.; Donoso, E.; hide

    2012-01-01

    After eight months of continuous observations, the Wide-field Infrared Survey Explorer (WISE) mapped the entire sky at 3.4 micron, 4.6 micron, 12 micron, and 22 micron. We have begun a dedicated WISE High Resolution Galaxy Atlas project to fully characterize large, nearby galaxies and produce a legacy image atlas and source catalog. Here we summarize the deconvolution techniques used to significantly improve the spatial resolution of WISE imaging, specifically designed to study the internal anatomy of nearby galaxies. As a case study, we present results for the galaxy NGC 1566, comparing the WISE enhanced-resolution image processing to that of Spitzer, Galaxy Evolution Explorer, and ground-based imaging. This is the first paper in a two-part series; results for a larger sample of nearby galaxies are presented in the second paper.

  18. A high resolution jet analysis for LEP

    International Nuclear Information System (INIS)

    Hariri, S.

    1992-11-01

    A high resolution multijet analysis of hadronic events produced in e + e - annihilation at a C.M.S. energy of 91.2 GeV is described. Hadronic events produced in e + e - annihilations are generated using the Monte Carlo program JETSET7.3 with its two options: Matrix Element (M.E.) and Parton Showers (P.S.). The shower option is used with its default parameter values while the M.E. option is used with an invariant mass cut Y CUT =0.01 instead of 0.02. This choice ensures a better continuity in the evolution of the event shape variables. (K.A.) 3 refs.; 26 figs.; 1 tab

  19. High Resolution Displays Using NCAP Liquid Crystals

    Science.gov (United States)

    Macknick, A. Brian; Jones, Phil; White, Larry

    1989-07-01

    Nematic curvilinear aligned phase (NCAP) liquid crystals have been found useful for high information content video displays. NCAP materials are liquid crystals which have been encapsulated in a polymer matrix and which have a light transmission which is variable with applied electric fields. Because NCAP materials do not require polarizers, their on-state transmission is substantially better than twisted nematic cells. All dimensional tolerances are locked in during the encapsulation process and hence there are no critical sealing or spacing issues. By controlling the polymer/liquid crystal morphology, switching speeds of NCAP materials have been significantly improved over twisted nematic systems. Recent work has combined active matrix addressing with NCAP materials. Active matrices, such as thin film transistors, have given displays of high resolution. The paper will discuss the advantages of NCAP materials specifically designed for operation at video rates on transistor arrays; applications for both backlit and projection displays will be discussed.

  20. High resolution VUV facility at INDUS-1

    International Nuclear Information System (INIS)

    Krishnamurty, G.; Saraswathy, P.; Rao, P.M.R.; Mishra, A.P.; Kartha, V.B.

    1993-01-01

    Synchrotron radiation (SR) generated in the electron storage rings is an unique source for the study of atomic and molecular spectroscopy especially in the vacuum ultra violet region. Realizing the potential of this light source, efforts are in progress to develop a beamline facility at INDUS-1 to carry out high resolution atomic and molecular spectroscopy. This beam line consists of a fore-optic which is a combination of three cylindrical mirrors. The mirrors are so chosen that SR beam having a 60 mrad (horizontal) x 6 mrad (vertical) divergence is focussed onto a slit of a 6.65 metre off-plane spectrometer in Eagle Mount equipped with horizontal slit and vertical dispersion. The design of the various components of the beam line is completed. It is decided to build the spectrometer as per the requirements of the user community. Details of the various aspects of the beam line will be presented. (author). 3 figs

  1. High-resolution CT of airway reactivity

    International Nuclear Information System (INIS)

    Herold, C.J.; Brown, R.H.; Hirshman, C.A.; Mitzner, W.; Zerhouni, E.A.

    1990-01-01

    Assessment of airway reactivity has generally been limited to experimental nonimaging models. This authors of this paper used high-resolution CT (HRCT) to evaluate airway reactivity and to calculate airway resistance (Raw) compared with lung resistance (RL). Ten anesthetized and ventilated dogs were investigated with HRCT (10 contiguous 2-mm sections through the lower lung lobes) during control state, following aerosol histamine challenge, and following posthistamine hyperinflation. The HRCT scans were digitized, and areas of 10 airways per dog (diameter, 1-10 mm) were measured with a computer edging process. Changes in airway area and Raw (calculated by 1/[area] 2 ) were measured. RL was assessed separately, following the same protocol. Data were analyzed by use of a paired t-test with significance at p < .05

  2. High resolution crystal calorimetry at LHC

    International Nuclear Information System (INIS)

    Schneegans, M.; Ferrere, D.; Lebeau, M.; Vivargent, M.

    1991-01-01

    The search for Higgs bosons above Lep200 reach could be one of the main tasks of the future pp and ee colliders. In the intermediate mass region, and in particular in the range 80-140 GeV/c 2 , only the 2-photon decay mode of a Higgs produced inclusively or in association with a W, gives a good chance of observation. A 'dedicated' very high resolution calorimeter with photon angle reconstruction and pion identification capability should detect a Higgs signal with high probability. A crystal calorimeter can be considered as a conservative approach to such a detector, since a large design and operation experience already exists. The extensive R and D needed for finding a dense, fast and radiation hard crystal, is under way. Guide-lines for designing an optimum calorimeter for LHC are discussed and preliminary configurations are given. (author) 7 refs., 3 figs., 2 tabs

  3. High resolution tomography using analog coding

    International Nuclear Information System (INIS)

    Brownell, G.L.; Burnham, C.A.; Chesler, D.A.

    1985-01-01

    As part of a 30-year program in the development of positron instrumentation, the authors have developed a high resolution bismuth germanate (BGO) ring tomography (PCR) employing 360 detectors and 90 photomultiplier tubes for one plane. The detectors are shaped as trapezoid and are 4 mm wide at the front end. When assembled, they form an essentially continuous cylindrical detector. Light from a scintillation in the detector is viewed through a cylindrical light pipe by the photomultiplier tubes. By use of an analog coding scheme, the detector emitting light is identified from the phototube signals. In effect, each phototube can identify four crystals. PCR is designed as a static device and does not use interpolative motion. This results in considerable advantage when performing dynamic studies. PCR is the positron tomography analog of the γ-camera widely used in nuclear medicine

  4. High-resolution CT of otosclerosis

    International Nuclear Information System (INIS)

    Dewen, Yang; Kodama, Takao; Tono, Tetsuya; Ochiai, Reiji; Kiyomizu, Kensuke; Suzuki, Yukiko; Yano, Takanori; Watanabe, Katsushi

    1997-01-01

    High-resolution CT (HRCT) scans of thirty-two patients (60 ears) with the clinical diagnosis of fenestral otosclerosis were evaluated retrospectively. HRCT was performed with 1-mm-thick targeted sections and 1-mm (36 ears) or 0.5-mm (10 ears) intervals in the semiaxial projection. Seven patients (14 ears) underwent helical scanning with a 1-mm slice thickness and 1-mm/sec table speed. Forty-five ears (75%) were found to have one or more otospongiotic or otosclerotic foci on HRCT. In most instances (30 ears), the otospongiotic foci were found in the region of the fissula ante fenestram. No significant correlations between CT findings and air conduction threshold were observed. We found a significant relationship between lesions of the labyrinthine capsule and sensorineural hearing loss. We conclude that HRCT is a valuable modality for diagnosing otosclerosis, especially when otospongiotic focus is detected. (author)

  5. High resolution CT in pulmonary sarcoidosis

    International Nuclear Information System (INIS)

    Spina, Juan C.; Curros, Marisela L.; Gomez, M.; Gonzalez, A.; Chacon, Carolina; Guerendiain, G.

    2000-01-01

    Objectives: To establish the particular advantages of High Resolution CT (HRCT) for the diagnosis of pulmonary sarcoidosis. Material and Methods: A series of fourteen patients, (4 men and 10 women; mean age 44,5 years) with thoracic sarcoidosis. All patients were studied using HRCT and diagnosis was confirmed for each case. Confidence intervals were obtained for different disease manifestations. Results: The most common findings were: lymph node enlargement (n=14 patients), pulmonary nodules (n=13), thickening of septa (n=6), peribronquial vascular thickening (n=5) pulmonary pseudo mass (n=5) and signs of fibrosis (n=4). The stage most commonly observed was stage II. It is worth noting that no cases of pleural effusion or cavitations of pulmonary lesions were observed. Conclusions: In this series, confidence interval overlapping for lymph node enlargement, single pulmonary nodules and septum thickening, allows to infer that their presence in a young adult, with few clinical symptoms, forces to rule out first the possibility of sarcoidosis. (author)

  6. Improved methods for high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, J.R.

    1987-04-01

    Existing methods of making support films for high resolution transmission electron microscopy are investigated and novel methods are developed. Existing methods of fabricating fenestrated, metal reinforced specimen supports (microgrids) are evaluated for their potential to reduce beam induced movement of monolamellar crystals of C/sub 44/H/sub 90/ paraffin supported on thin carbon films. Improved methods of producing hydrophobic carbon films by vacuum evaporation, and improved methods of depositing well ordered monolamellar paraffin crystals on carbon films are developed. A novel technique for vacuum evaporation of metals is described which is used to reinforce microgrids. A technique is also developed to bond thin carbon films to microgrids with a polymer bonding agent. Unique biochemical methods are described to accomplish site specific covalent modification of membrane proteins. Protocols are given which covalently convert the carboxy terminus of papain cleaved bacteriorhodopsin to a free thiol. 53 refs., 19 figs., 1 tab.

  7. High resolution infrared spectroscopy of symbiotic stars

    International Nuclear Information System (INIS)

    Bensammar, S.

    1989-01-01

    We report here very early results of high resolution (5x10 3 - 4x10 4 ) infrared spectroscopy (1 - 2.5 μm) of different symbiotic stars (T CrB, RW Hya, CI Cyg, PU Vul) observed with the Fourier Transform Spectrometer of the 3.60m Canada France Hawaii Telescope. These stars are usually considered as interacting binaries and only little details are known about the nature of their cool component. CO absorption lines are detected for the four stars. Very different profiles of hydrogen Brackett γ and helium 10830 A lines are shown for CI Cyg observed at different phases, while Pu Vul shows very intense emission lines

  8. GRANULOMETRIC MAPS FROM HIGH RESOLUTION SATELLITE IMAGES

    Directory of Open Access Journals (Sweden)

    Catherine Mering

    2011-05-01

    Full Text Available A new method of land cover mapping from satellite images using granulometric analysis is presented here. Discontinuous landscapes such as steppian bushes of semi arid regions and recently growing urban settlements are especially concerned by this study. Spatial organisations of the land cover are quantified by means of the size distribution analysis of the land cover units extracted from high resolution remotely sensed images. A granulometric map is built by automatic classification of every pixel of the image according to the granulometric density inside a sliding neighbourhood. Granulometric mapping brings some advantages over traditional thematic mapping by remote sensing by focusing on fine spatial events and small changes in one peculiar category of the landscape.

  9. Preparation of very small point sources for high resolution radiography

    International Nuclear Information System (INIS)

    Case, F.N.

    1976-01-01

    The need for very small point sources of high specific activity 192 Ir, 169 Yb, 170 Tm, and 60 Co in non-destructive testing has motivated the development of techniques for the fabrication of these sources. To prepare 192 Ir point sources for use in examination of tube sheet welds in LMFBR heat exchangers, 191 Ir enriched to greater than 90 percent was melted in a helium blanketed arc to form spheres as small as 0.38 mm in diameter. Methods were developed to form the roughly spherical shaped arc product into nearly symmetrical spheres that could be used for high resolution radiography. Similar methods were used for spherical shaped sources of 169 Yb and 170 Tm. The oxides were arc melted to form rough spheres followed by grinding to precise dimensions, neutron irradiation of the spheres at a flux of 2 to 3 x 10 15 nv, and use of enriched 168 Yb to provide the maximum specific activity. Cobalt-60 with a specific activity of greater than 1100 Ci/g was prepared by processing 59 Co that had been neutron irradiated to nearly complete burnup of the 59 Co target to produce 60 Co, 61 Ni, and 62 Ni. Ion exchange methods were used to separate the cobalt from the nickel. The cobalt was reduced to metal by plating either onto aluminum foil which was dissolved away from the cobalt plate, or by plating onto mercury to prepare amalgam that could be easily formed into a pellet of cobalt with exclusion of the mercury. Both methods are discussed

  10. Integrated High Resolution Monitoring of Mediterranean vegetation

    Science.gov (United States)

    Cesaraccio, Carla; Piga, Alessandra; Ventura, Andrea; Arca, Angelo; Duce, Pierpaolo; Mereu, Simone

    2017-04-01

    The study of the vegetation features in a complex and highly vulnerable ecosystems, such as Mediterranean maquis, leads to the need of using continuous monitoring systems at high spatial and temporal resolution, for a better interpretation of the mechanisms of phenological and eco-physiological processes. Near-surface remote sensing techniques are used to quantify, at high temporal resolution, and with a certain degree of spatial integration, the seasonal variations of the surface optical and radiometric properties. In recent decades, the design and implementation of global monitoring networks involved the use of non-destructive and/or cheaper approaches such as (i) continuous surface fluxes measurement stations, (ii) phenological observation networks, and (iii) measurement of temporal and spatial variations of the vegetation spectral properties. In this work preliminary results from the ECO-SCALE (Integrated High Resolution Monitoring of Mediterranean vegetation) project are reported. The project was manly aimed to develop an integrated system for environmental monitoring based on digital photography, hyperspectral radiometry , and micrometeorological techniques during three years of experimentation (2013-2016) in a Mediterranean site of Italy (Capo Caccia, Alghero). The main results concerned the analysis of chromatic coordinates indices from digital images, to characterized the phenological patterns for typical shrubland species, determining start and duration of the growing season, and the physiological status in relation to different environmental drought conditions; then the seasonal patterns of canopy phenology, was compared to NEE (Net Ecosystem Exchange) patterns, showing similarities. However, maximum values of NEE and ER (Ecosystem respiration), and short term variation, seemed mainly tuned by inter annual pattern of meteorological variables, in particular of temperature recorded in the months preceding the vegetation green-up. Finally, green signals

  11. Dynamic high resolution imaging of rats

    International Nuclear Information System (INIS)

    Miyaoka, R.S.; Lewellen, T.K.; Bice, A.N.

    1990-01-01

    A positron emission tomography with the sensitivity and resolution to do dynamic imaging of rats would be an invaluable tool for biological researchers. In this paper, the authors determine the biological criteria for dynamic positron emission imaging of rats. To be useful, 3 mm isotropic resolution and 2-3 second time binning were necessary characteristics for such a dedicated tomograph. A single plane in which two objects of interest could be imaged simultaneously was considered acceptable. Multi-layered detector designs were evaluated as a possible solution to the dynamic imaging and high resolution imaging requirements. The University of Washington photon history generator was used to generate data to investigate a tomograph's sensitivity to true, scattered and random coincidences for varying detector ring diameters. Intrinsic spatial uniformity advantages of multi-layered detector designs over conventional detector designs were investigated using a Monte Carlo program. As a result, a modular three layered detector prototype is being developed. A module will consist of a layer of five 3.5 mm wide crystals and two layers of six 2.5 mm wide crystals. The authors believe adequate sampling can be achieved with a stationary detector system using these modules. Economical crystal decoding strategies have been investigated and simulations have been run to investigate optimum light channeling methods for block decoding strategies. An analog block decoding method has been proposed and will be experimentally evaluated to determine whether it can provide the desired performance

  12. High resolution computed tomography of positron emitters

    International Nuclear Information System (INIS)

    Derenzo, S.E.; Budinger, T.F.; Cahoon, J.L.; Huesman, R.H.; Jackson, H.G.

    1976-10-01

    High resolution computed transaxial radionuclide tomography has been performed on phantoms containing positron-emitting isotopes. The imaging system consisted of two opposing groups of eight NaI(Tl) crystals 8 mm x 30 mm x 50 mm deep and the phantoms were rotated to measure coincident events along 8960 projection integrals as they would be measured by a 280-crystal ring system now under construction. The spatial resolution in the reconstructed images is 7.5 mm FWHM at the center of the ring and approximately 11 mm FWHM at a radius of 10 cm. We present measurements of imaging and background rates under various operating conditions. Based on these measurements, the full 280-crystal system will image 10,000 events per sec with 400 μCi in a section 1 cm thick and 20 cm in diameter. We show that 1.5 million events are sufficient to reliably image 3.5-mm hot spots with 14-mm center-to-center spacing and isolated 9-mm diameter cold spots in phantoms 15 to 20 cm in diameter

  13. High speed, High resolution terahertz spectrometers

    International Nuclear Information System (INIS)

    Kim, Youngchan; Yee, Dae Su; Yi, Miwoo; Ahn, Jaewook

    2008-01-01

    A variety of sources and methods have been developed for terahertz spectroscopy during almost two decades. Terahertz time domain spectroscopy (THz TDS)has attracted particular attention as a basic measurement method in the fields of THz science and technology. Recently, asynchronous optical sampling (AOS)THz TDS has been demonstrated, featuring rapid data acquisition and a high spectral resolution. Also, terahertz frequency comb spectroscopy (TFCS)possesses attractive features for high precision terahertz spectroscopy. In this presentation, we report on these two types of terahertz spectrometer. Our high speed, high resolution terahertz spectrometer is demonstrated using two mode locked femtosecond lasers with slightly different repetition frequencies without a mechanical delay stage. The repetition frequencies of the two femtosecond lasers are stabilized by use of two phase locked loops sharing the same reference oscillator. The time resolution of our terahertz spectrometer is measured using the cross correlation method to be 270 fs. AOS THz TDS is presented in Fig. 1, which shows a time domain waveform rapidly acquired on a 10ns time window. The inset shows a zoom into the signal with 100ps time window. The spectrum obtained by the fast Fourier Transformation (FFT)of the time domain waveform has a frequency resolution of 100MHz. The dependence of the signal to noise ratio (SNR)on the measurement time is also investigated

  14. High resolution CT of temporal bone trauma

    International Nuclear Information System (INIS)

    Youn, Eun Kyung

    1986-01-01

    Radiographic studies of the temporal bone following head trauma are indicated when there is cerebrospinal fluid otorrhea or rhinorrhoea, hearing loss, or facial nerve paralysis. Plain radiography displays only 17-30% of temporal bone fractures and pluridirectional tomography is both difficult to perform, particularly in the acutely ill patient, and less satisfactory for the demonstration of fine fractures. Consequently, high resolution CT is the imaging method of choice for the investigation of suspected temporal bone trauma and allows special resolution of fine bony detail comparable to that attainable by conventional tomography. Eight cases of temporal bone trauma examined at Korea General Hospital April 1985 through May 1986. The results were as follows: Seven patients (87%) suffered longitudinal fractures. In 6 patients who had purely conductive hearing loss, CT revealed various ossicular chain abnormality. In one patient who had neuro sensory hearing loss, CT demonstrated intract ossicular with a fracture nearing lateral wall of the lateral semicircular canal. In one patient who had mixed hearing loss, CT showed complex fracture.

  15. High resolution SETI: Experiences and prospects

    Science.gov (United States)

    Horowitz, Paul; Clubok, Ken

    Megachannel spectroscopy with sub-Hertz resolution constitutes an attractive strategy for a microwave search for extraterrestrial intelligence (SETI), assuming the transmission of a narrowband radiofrequency beacon. Such resolution matches the properties of the interstellar medium, and the necessary Doppler corrections provide a high degree of interference rejection. We have constructed a frequency-agile receiver with an FFT-based 8 megachannel digital spectrum analyzer, on-line signal recognition, and multithreshold archiving. We are using it to conduct a meridian transit search of the northern sky at the Harvard-Smithsonian 26-m antenna, with a second identical system scheduled to begin observations in Argentina this month. Successive 400 kHz spectra, at 0.05 Hz resolution, are searched for features characteristic of an intentional narrowband beacon transmission. These spectra are centered on guessable frequencies (such as λ21 cm), referenced successively to the local standard of rest, the galactic barycenter, and the cosmic blackbody rest frame. This search has rejected interference admirably, but is greatly limited both in total frequency coverage and sensitivity to signals other than carriers. We summarize five years of high resolution SETI at Harvard, in the context of answering the questions "How useful is narrowband SETI, how serious are its limitations, what can be done to circumvent them, and in what direction should SETI evolve?" Increasingly powerful signal processing hardware, combined with ever-higher memory densities, are particularly relevant, permitting the construction of compact and affordable gigachannel spectrum analyzers covering hundreds of megahertz of instantaneous bandwidth.

  16. High-resolution CCD imaging alternatives

    Science.gov (United States)

    Brown, D. L.; Acker, D. E.

    1992-08-01

    High resolution CCD color cameras have recently stimulated the interest of a large number of potential end-users for a wide range of practical applications. Real-time High Definition Television (HDTV) systems are now being used or considered for use in applications ranging from entertainment program origination through digital image storage to medical and scientific research. HDTV generation of electronic images offers significant cost and time-saving advantages over the use of film in such applications. Further in still image systems electronic image capture is faster and more efficient than conventional image scanners. The CCD still camera can capture 3-dimensional objects into the computing environment directly without having to shoot a picture on film develop it and then scan the image into a computer. 2. EXTENDING CCD TECHNOLOGY BEYOND BROADCAST Most standard production CCD sensor chips are made for broadcast-compatible systems. One popular CCD and the basis for this discussion offers arrays of roughly 750 x 580 picture elements (pixels) or a total array of approximately 435 pixels (see Fig. 1). FOR. A has developed a technique to increase the number of available pixels for a given image compared to that produced by the standard CCD itself. Using an inter-lined CCD with an overall spatial structure several times larger than the photo-sensitive sensor areas each of the CCD sensors is shifted in two dimensions in order to fill in spatial gaps between adjacent sensors.

  17. High resolution simultaneous measurements of airborne radionuclides

    International Nuclear Information System (INIS)

    Abe, T.; Yamaguchi, Y.; Tanaka, K.; Komura, K.

    2006-01-01

    High resolution (2-3 hrs) simultaneous measurements of airborne radionuclides, 212 Pb, 210 Pb and 7 Be, have been performed by using extremely low background Ge detectors at Ogoya Underground Laboratory. We have measured above radionuclides at three monitoring points viz, 1) Low Level Radioactivity Laboratory (LLRL) Kanazawa University, 2) Shishiku Plateau (640 m MSL) located about 8 km from LLRL to investigate vertical difference of activity levels, and 3) Hegura Island (10 m MSL) located about 50 km from Noto Peninsula in the Sea of Japan to evaluate the influences of Asian continent or mainland of Japan on the variation to the activity levels. Variations of short-lived 212 Pb concentration showed noticeable time lags between at LLRL and at Shishiku Plateau. These time lags might be caused by change of height of a planetary boundary layer. On the contrary, variations of long-lived 210 Pb and 7 Be showed simultaneity at three locations because of homogeneity of these concentrations all over the area. (author)

  18. High-resolution X-ray television and high-resolution video recorders

    International Nuclear Information System (INIS)

    Haendle, J.; Horbaschek, H.; Alexandrescu, M.

    1977-01-01

    The improved transmission properties of the high-resolution X-ray television chain described here make it possible to transmit more information per television image. The resolution in the fluoroscopic image, which is visually determined, depends on the dose rate and the inertia of the television pick-up tube. This connection is discussed. In the last few years, video recorders have been increasingly used in X-ray diagnostics. The video recorder is a further quality-limiting element in X-ray television. The development of function patterns of high-resolution magnetic video recorders shows that this quality drop may be largely overcome. The influence of electrical band width and number of lines on the resolution in the X-ray television image stored is explained in more detail. (orig.) [de

  19. Processing method for high resolution monochromator

    International Nuclear Information System (INIS)

    Kiriyama, Koji; Mitsui, Takaya

    2006-12-01

    A processing method for high resolution monochromator (HRM) has been developed at Japanese Atomic Energy Agency/Quantum Beam Science Directorate/Synchrotron Radiation Research unit at SPring-8. For manufacturing a HRM, a sophisticated slicing machine and X-ray diffractometer have been installed for shaping a crystal ingot and orienting precisely the surface of a crystal ingot, respectively. The specification of the slicing machine is following; Maximum size of a diamond blade is φ 350mm in diameter, φ 38.1mm in the spindle diameter, and 2mm in thickness. A large crystal such as an ingot with 100mm in diameter, 200mm in length can be cut. Thin crystal samples such as a wafer can be also cut using by another sample holder. Working distance of a main shaft with the direction perpendicular to working table in the machine is 350mm at maximum. Smallest resolution of the main shaft with directions of front-and-back and top-and-bottom are 0.001mm read by a digital encoder. 2mm/min can set for cutting samples in the forward direction. For orienting crystal faces relative to the blade direction adjustment, a one-circle goniometer and 2-circle segment are equipped on the working table in the machine. A rotation and a tilt of the stage can be done by manual operation. Digital encoder in a turn stage is furnished and has angle resolution of less than 0.01 degrees. In addition, a hand drill as a supporting device for detailed processing of crystal is prepared. Then, an ideal crystal face can be cut from crystal samples within an accuracy of about 0.01 degrees. By installation of these devices, a high energy resolution monochromator crystal for inelastic x-ray scattering and a beam collimator are got in hand and are expected to be used for nanotechnology studies. (author)

  20. Zeolites - a high resolution electron microscopy study

    International Nuclear Information System (INIS)

    Alfredsson, V.

    1994-10-01

    High resolution transmission electron microscopy (HRTEM) has been used to investigate a number of zeolites (EMT, FAU, LTL, MFI and MOR) and a member of the mesoporous M41S family. The electron optical artefact, manifested as a dark spot in the projected centre of the large zeolite channels, caused by insufficient transfer of certain reflections in the objective lens has been explained. The artefact severely hinders observation of materials confined in the zeolite channels and cavities. It is shown how to circumvent the artefact problem and how to image confined materials in spite of disturbance caused by the artefact. Image processing by means of a Wiener filter has been applied for removal of the artefact. The detailed surface structure of FAU has been investigated. Comparison of experimental micrographs with images simulated using different surface models indicates that the surface can be terminated in different ways depending on synthesis methods. The dealuminated form of FAU (USY) is covered by an amorphous region. Platinum incorporated in FAU has a preponderance to aggregate in the (111) twin planes, probably due to a local difference in cage structure with more spacious cages. It is shown that platinum is intra-zeolitic as opposed to being located on the external surface of the zeolite crystal. This could be deduced from tomography of ultra-thin sections among observations. HRTEM studies of the mesoporous MCM-41 show that the pores have a hexagonal shape and also supports the mechanistic model proposed which involves a cooperative formation of a mesophase including the silicate species as well as the surfactant. 66 refs, 24 figs

  1. High-resolution downscaling for hydrological management

    Science.gov (United States)

    Ulbrich, Uwe; Rust, Henning; Meredith, Edmund; Kpogo-Nuwoklo, Komlan; Vagenas, Christos

    2017-04-01

    Hydrological modellers and water managers require high-resolution climate data to model regional hydrologies and how these may respond to future changes in the large-scale climate. The ability to successfully model such changes and, by extension, critical infrastructure planning is often impeded by a lack of suitable climate data. This typically takes the form of too-coarse data from climate models, which are not sufficiently detailed in either space or time to be able to support water management decisions and hydrological research. BINGO (Bringing INnovation in onGOing water management; ) aims to bridge the gap between the needs of hydrological modellers and planners, and the currently available range of climate data, with the overarching aim of providing adaptation strategies for climate change-related challenges. Producing the kilometre- and sub-daily-scale climate data needed by hydrologists through continuous simulations is generally computationally infeasible. To circumvent this hurdle, we adopt a two-pronged approach involving (1) selective dynamical downscaling and (2) conditional stochastic weather generators, with the former presented here. We take an event-based approach to downscaling in order to achieve the kilometre-scale input needed by hydrological modellers. Computational expenses are minimized by identifying extremal weather patterns for each BINGO research site in lower-resolution simulations and then only downscaling to the kilometre-scale (convection permitting) those events during which such patterns occur. Here we (1) outline the methodology behind the selection of the events, and (2) compare the modelled precipitation distribution and variability (preconditioned on the extremal weather patterns) with that found in observations.

  2. Toward high-resolution optoelectronic retinal prosthesis

    Science.gov (United States)

    Palanker, Daniel; Huie, Philip; Vankov, Alexander; Asher, Alon; Baccus, Steven

    2005-04-01

    It has been already demonstrated that electrical stimulation of retina can produce visual percepts in blind patients suffering from macular degeneration and retinitis pigmentosa. Current retinal implants provide very low resolution (just a few electrodes), while several thousand pixels are required for functional restoration of sight. We present a design of the optoelectronic retinal prosthetic system that can activate a retinal stimulating array with pixel density up to 2,500 pix/mm2 (geometrically corresponding to a visual acuity of 20/80), and allows for natural eye scanning rather than scanning with a head-mounted camera. The system operates similarly to "virtual reality" imaging devices used in military and medical applications. An image from a video camera is projected by a goggle-mounted infrared LED-LCD display onto the retina, activating an array of powered photodiodes in the retinal implant. Such a system provides a broad field of vision by allowing for natural eye scanning. The goggles are transparent to visible light, thus allowing for simultaneous utilization of remaining natural vision along with prosthetic stimulation. Optical control of the implant allows for simple adjustment of image processing algorithms and for learning. A major prerequisite for high resolution stimulation is the proximity of neural cells to the stimulation sites. This can be achieved with sub-retinal implants constructed in a manner that directs migration of retinal cells to target areas. Two basic implant geometries are described: perforated membranes and protruding electrode arrays. Possibility of the tactile neural stimulation is also examined.

  3. High-resolution phylogenetic microbial community profiling

    Energy Technology Data Exchange (ETDEWEB)

    Singer, Esther; Coleman-Derr, Devin; Bowman, Brett; Schwientek, Patrick; Clum, Alicia; Copeland, Alex; Ciobanu, Doina; Cheng, Jan-Fang; Gies, Esther; Hallam, Steve; Tringe, Susannah; Woyke, Tanja

    2014-03-17

    The representation of bacterial and archaeal genome sequences is strongly biased towards cultivated organisms, which belong to merely four phylogenetic groups. Functional information and inter-phylum level relationships are still largely underexplored for candidate phyla, which are often referred to as microbial dark matter. Furthermore, a large portion of the 16S rRNA gene records in the GenBank database are labeled as environmental samples and unclassified, which is in part due to low read accuracy, potential chimeric sequences produced during PCR amplifications and the low resolution of short amplicons. In order to improve the phylogenetic classification of novel species and advance our knowledge of the ecosystem function of uncultivated microorganisms, high-throughput full length 16S rRNA gene sequencing methodologies with reduced biases are needed. We evaluated the performance of PacBio single-molecule real-time (SMRT) sequencing in high-resolution phylogenetic microbial community profiling. For this purpose, we compared PacBio and Illumina metagenomic shotgun and 16S rRNA gene sequencing of a mock community as well as of an environmental sample from Sakinaw Lake, British Columbia. Sakinaw Lake is known to contain a large age of microbial species from candidate phyla. Sequencing results show that community structure based on PacBio shotgun and 16S rRNA gene sequences is highly similar in both the mock and the environmental communities. Resolution power and community representation accuracy from SMRT sequencing data appeared to be independent of GC content of microbial genomes and was higher when compared to Illumina-based metagenome shotgun and 16S rRNA gene (iTag) sequences, e.g. full-length sequencing resolved all 23 OTUs in the mock community, while iTags did not resolve closely related species. SMRT sequencing hence offers various potential benefits when characterizing uncharted microbial communities.

  4. HIGH RESOLUTION AIRBORNE SHALLOW WATER MAPPING

    Directory of Open Access Journals (Sweden)

    F. Steinbacher

    2012-07-01

    Full Text Available In order to meet the requirements of the European Water Framework Directive (EU-WFD, authorities face the problem of repeatedly performing area-wide surveying of all kinds of inland waters. Especially for mid-sized or small rivers this is a considerable challenge imposing insurmountable logistical efforts and costs. It is therefore investigated if large-scale surveying of a river system on an operational basis is feasible by employing airborne hydrographic laser scanning. In cooperation with the Bavarian Water Authority (WWA Weilheim a pilot project was initiated by the Unit of Hydraulic Engineering at the University of Innsbruck and RIEGL Laser Measurement Systems exploiting the possibilities of a new LIDAR measurement system with high spatial resolution and high measurement rate to capture about 70 km of riverbed and foreland for the river Loisach in Bavaria/Germany and the estuary and parts of the shoreline (about 40km in length of lake Ammersee. The entire area surveyed was referenced to classic terrestrial cross-section surveys with the aim to derive products for the monitoring and managing needs of the inland water bodies forced by the EU-WFD. The survey was performed in July 2011 by helicopter and airplane and took 3 days in total. In addition, high resolution areal images were taken to provide an optical reference, offering a wide range of possibilities on further research, monitoring, and managing responsibilities. The operating altitude was about 500 m to maintain eye-safety, even for the aided eye, the airspeed was about 55 kts for the helicopter and 75 kts for the aircraft. The helicopter was used in the alpine regions while the fixed wing aircraft was used in the plains and the urban area, using appropriate scan rates to receive evenly distributed point clouds. The resulting point density ranged from 10 to 25 points per square meter. By carefully selecting days with optimum water quality, satisfactory penetration down to the river

  5. High Resolution Airborne Shallow Water Mapping

    Science.gov (United States)

    Steinbacher, F.; Pfennigbauer, M.; Aufleger, M.; Ullrich, A.

    2012-07-01

    In order to meet the requirements of the European Water Framework Directive (EU-WFD), authorities face the problem of repeatedly performing area-wide surveying of all kinds of inland waters. Especially for mid-sized or small rivers this is a considerable challenge imposing insurmountable logistical efforts and costs. It is therefore investigated if large-scale surveying of a river system on an operational basis is feasible by employing airborne hydrographic laser scanning. In cooperation with the Bavarian Water Authority (WWA Weilheim) a pilot project was initiated by the Unit of Hydraulic Engineering at the University of Innsbruck and RIEGL Laser Measurement Systems exploiting the possibilities of a new LIDAR measurement system with high spatial resolution and high measurement rate to capture about 70 km of riverbed and foreland for the river Loisach in Bavaria/Germany and the estuary and parts of the shoreline (about 40km in length) of lake Ammersee. The entire area surveyed was referenced to classic terrestrial cross-section surveys with the aim to derive products for the monitoring and managing needs of the inland water bodies forced by the EU-WFD. The survey was performed in July 2011 by helicopter and airplane and took 3 days in total. In addition, high resolution areal images were taken to provide an optical reference, offering a wide range of possibilities on further research, monitoring, and managing responsibilities. The operating altitude was about 500 m to maintain eye-safety, even for the aided eye, the airspeed was about 55 kts for the helicopter and 75 kts for the aircraft. The helicopter was used in the alpine regions while the fixed wing aircraft was used in the plains and the urban area, using appropriate scan rates to receive evenly distributed point clouds. The resulting point density ranged from 10 to 25 points per square meter. By carefully selecting days with optimum water quality, satisfactory penetration down to the river bed was achieved

  6. High resolution time integration for SN radiation transport

    International Nuclear Information System (INIS)

    Thoreson, Greg; McClarren, Ryan G.; Chang, Jae H.

    2009-01-01

    First-order, second-order, and high resolution time discretization schemes are implemented and studied for the discrete ordinates (S N ) equations. The high resolution method employs a rate of convergence better than first-order, but also suppresses artificial oscillations introduced by second-order schemes in hyperbolic partial differential equations. The high resolution method achieves these properties by nonlinearly adapting the time stencil to use a first-order method in regions where oscillations could be created. We employ a quasi-linear solution scheme to solve the nonlinear equations that arise from the high resolution method. All three methods were compared for accuracy and convergence rates. For non-absorbing problems, both second-order and high resolution converged to the same solution as the first-order with better convergence rates. High resolution is more accurate than first-order and matches or exceeds the second-order method

  7. A high resolution solar atlas for fluorescence calculations

    Science.gov (United States)

    Hearn, M. F.; Ohlmacher, J. T.; Schleicher, D. G.

    1983-01-01

    The characteristics required of a solar atlas to be used for studying the fluorescence process in comets are examined. Several sources of low resolution data were combined to provide an absolutely calibrated spectrum from 2250 A to 7000A. Three different sources of high resolution data were also used to cover this same spectral range. The low resolution data were then used to put each high resolution spectrum on an absolute scale. The three high resolution spectra were then combined in their overlap regions to produce a single, absolutely calibrated high resolution spectrum over the entire spectral range.

  8. High resolution time integration for Sn radiation transport

    International Nuclear Information System (INIS)

    Thoreson, Greg; McClarren, Ryan G.; Chang, Jae H.

    2008-01-01

    First order, second order and high resolution time discretization schemes are implemented and studied for the S n equations. The high resolution method employs a rate of convergence better than first order, but also suppresses artificial oscillations introduced by second order schemes in hyperbolic differential equations. All three methods were compared for accuracy and convergence rates. For non-absorbing problems, both second order and high resolution converged to the same solution as the first order with better convergence rates. High resolution is more accurate than first order and matches or exceeds the second order method. (authors)

  9. The implementation of sea ice model on a regional high-resolution scale

    Science.gov (United States)

    Prasad, Siva; Zakharov, Igor; Bobby, Pradeep; McGuire, Peter

    2015-09-01

    The availability of high-resolution atmospheric/ocean forecast models, satellite data and access to high-performance computing clusters have provided capability to build high-resolution models for regional ice condition simulation. The paper describes the implementation of the Los Alamos sea ice model (CICE) on a regional scale at high resolution. The advantage of the model is its ability to include oceanographic parameters (e.g., currents) to provide accurate results. The sea ice simulation was performed over Baffin Bay and the Labrador Sea to retrieve important parameters such as ice concentration, thickness, ridging, and drift. Two different forcing models, one with low resolution and another with a high resolution, were used for the estimation of sensitivity of model results. Sea ice behavior over 7 years was simulated to analyze ice formation, melting, and conditions in the region. Validation was based on comparing model results with remote sensing data. The simulated ice concentration correlated well with Advanced Microwave Scanning Radiometer for EOS (AMSR-E) and Ocean and Sea Ice Satellite Application Facility (OSI-SAF) data. Visual comparison of ice thickness trends estimated from the Soil Moisture and Ocean Salinity satellite (SMOS) agreed with the simulation for year 2010-2011.

  10. High Resolution Global Electrical Conductivity Variations in the Earth's Mantle

    Science.gov (United States)

    Kelbert, A.; Sun, J.; Egbert, G. D.

    2013-12-01

    Electrical conductivity of the Earth's mantle is a valuable constraint on the water content and melting processes. In Kelbert et al. (2009), we obtained the first global inverse model of electrical conductivity in the mantle capable of providing constraints on the lateral variations in mantle water content. However, in doing so we had to compromise on the problem complexity by using the historically very primitive ionospheric and magnetospheric source assumptions. In particular, possible model contamination by the auroral current systems had greatly restricted our use of available data. We have now addressed this problem by inverting for the external sources along with the electrical conductivity variations. In this study, we still focus primarily on long period data that are dominated by quasi-zonal source fields. The improved understanding of the ionospheric sources allows us to invert the magnetic fields directly, without a correction for the source and/or the use of transfer functions. It allows us to extend the period range of available data to 1.2 days - 102 days, achieving better sensitivity to the upper mantle and transition zone structures. Finally, once the source effects in the data are accounted for, a much larger subset of observatories may be used in the electrical conductivity inversion. Here, we use full magnetic fields at 207 geomagnetic observatories, which include mid-latitude, equatorial and high latitude data. Observatory hourly means from the years 1958-2010 are employed. The improved quality and spatial distribution of the data set, as well as the high resolution modeling and inversion using degree and order 40 spherical harmonics mapped to a 2x2 degree lateral grid, all contribute to the much improved resolution of our models, representing a conceptual step forward in global electromagnetic sounding. We present a fully three-dimensional, global electrical conductivity model of the Earth's mantle as inferred from ground geomagnetic

  11. High dimensional and high resolution pulse sequences for backbone resonance assignment of intrinsically disordered proteins

    Energy Technology Data Exchange (ETDEWEB)

    Zawadzka-Kazimierczuk, Anna; Kozminski, Wiktor, E-mail: kozmin@chem.uw.edu.pl [University of Warsaw, Faculty of Chemistry (Poland); Sanderova, Hana; Krasny, Libor [Institute of Microbiology, Academy of Sciences of the Czech Republic, Laboratory of Molecular Genetics of Bacteria, Department of Bacteriology (Czech Republic)

    2012-04-15

    Four novel 5D (HACA(N)CONH, HNCOCACB, (HACA)CON(CA)CONH, (H)NCO(NCA)CONH), and one 6D ((H)NCO(N)CACONH) NMR pulse sequences are proposed. The new experiments employ non-uniform sampling that enables achieving high resolution in indirectly detected dimensions. The experiments facilitate resonance assignment of intrinsically disordered proteins. The novel pulse sequences were successfully tested using {delta} subunit (20 kDa) of Bacillus subtilis RNA polymerase that has an 81-amino acid disordered part containing various repetitive sequences.

  12. Scalable Algorithms for Large High-Resolution Terrain Data

    DEFF Research Database (Denmark)

    Mølhave, Thomas; Agarwal, Pankaj K.; Arge, Lars Allan

    2010-01-01

    In this paper we demonstrate that the technology required to perform typical GIS computations on very large high-resolution terrain models has matured enough to be ready for use by practitioners. We also demonstrate the impact that high-resolution data has on common problems. To our knowledge, so...

  13. High-resolution X-ray diffraction studies of multilayers

    DEFF Research Database (Denmark)

    Christensen, Finn Erland; Hornstrup, Allan; Schnopper, H. W.

    1988-01-01

    High-resolution X-ray diffraction studies of the perfection of state-of-the-art multilayers are presented. Data were obtained using a triple-axis perfect-crystal X-ray diffractometer. Measurements reveal large-scale figure errors in the substrate. A high-resolution triple-axis set up is required...

  14. Achieving sensitive, high-resolution laser spectroscopy at CRIS

    Energy Technology Data Exchange (ETDEWEB)

    Groote, R. P. de [Instituut voor Kern- en Stralingsfysica, KU Leuven (Belgium); Lynch, K. M., E-mail: kara.marie.lynch@cern.ch [EP Department, CERN, ISOLDE (Switzerland); Wilkins, S. G. [The University of Manchester, School of Physics and Astronomy (United Kingdom); Collaboration: the CRIS collaboration

    2017-11-15

    The Collinear Resonance Ionization Spectroscopy (CRIS) experiment, located at the ISOLDE facility, has recently performed high-resolution laser spectroscopy, with linewidths down to 20 MHz. In this article, we present the modifications to the beam line and the newly-installed laser systems that have made sensitive, high-resolution measurements possible. Highlights of recent experimental campaigns are presented.

  15. High resolution UV spectroscopy and laser-focused nanofabrication

    NARCIS (Netherlands)

    Myszkiewicz, G.

    2005-01-01

    This thesis combines two at first glance different techniques: High Resolution Laser Induced Fluorescence Spectroscopy (LIF) of small aromatic molecules and Laser Focusing of atoms for Nanofabrication. The thesis starts with the introduction to the high resolution LIF technique of small aromatic

  16. High resolution NMR spectroscopy of synthetic polymers in bulk

    International Nuclear Information System (INIS)

    Komorski, R.A.

    1986-01-01

    The contents of this book are: Overview of high-resolution NMR of solid polymers; High-resolution NMR of glassy amorphous polymers; Carbon-13 solid-state NMR of semicrystalline polymers; Conformational analysis of polymers of solid-state NMR; High-resolution NMR studies of oriented polymers; High-resolution solid-state NMR of protons in polymers; and Deuterium NMR of solid polymers. This work brings together the various approaches for high-resolution NMR studies of bulk polymers into one volume. Heavy emphasis is, of course, given to 13C NMR studies both above and below Tg. Standard high-power pulse and wide-line techniques are not covered

  17. High resolution neutron spectroscopy - a tool for the investigation of dynamics of polymers and soft matter

    International Nuclear Information System (INIS)

    Monkenbusch, M.; Richter, D.

    2007-01-01

    Neutron scattering, with the ability to vary the contrast of molecular items by hydrogen/deuterium exchanges, is an invaluable tool for soft matter research. Besides the structural information on the mesoscopic scale that is obtained by diffraction methods like small angle neutron scattering, the slow dynamics of molecular motion on mesoscopic scale is accessible by high resolution neutron spectroscopy. The basic features of neutron backscattering spectroscopy, and in particular neutron spin-echo spectroscopy, are presented, in combination with illustrations of results from polymer melt dynamics to protein dynamics which are obtained by these techniques. (authors)

  18. A Forward-Looking High-Resolution GPR System

    National Research Council Canada - National Science Library

    Kositsky, Joel; Milanfar, Peyman

    1999-01-01

    A high-resolution ground penetrating radar (GPR) system was designed to help define the optimal radar parameters needed for the efficient standoff detection of buried and surface-laid antitank mines...

  19. High-resolution seismic wave propagation using local time stepping

    KAUST Repository

    Peter, Daniel; Rietmann, Max; Galvez, Percy; Ampuero, Jean Paul

    2017-01-01

    High-resolution seismic wave simulations often require local refinements in numerical meshes to accurately capture e.g. steep topography or complex fault geometry. Together with explicit time schemes, this dramatically reduces the global time step

  20. Impact of high resolution land surface initialization in Indian summer ...

    Indian Academy of Sciences (India)

    The direct impact of high resolution land surface initialization on the forecast bias in a regional climate model in recent years ... surface initialization using a regional climate model. ...... ization of the snow field in a cloud model; J. Clim. Appl.

  1. Textural Segmentation of High-Resolution Sidescan Sonar Images

    National Research Council Canada - National Science Library

    Kalcic, Maria; Bibee, Dale

    1995-01-01

    .... The high resolution of the 455 kHz sonar imagery also provides much information about the surficial bottom sediments, however their acoustic scattering properties are not well understood at high frequencies...

  2. NOAA High-Resolution Sea Surface Temperature (SST) Analysis Products

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This archive covers two high resolution sea surface temperature (SST) analysis products developed using an optimum interpolation (OI) technique. The analyses have a...

  3. Hurricane Satellite (HURSAT) from Advanced Very High Resolution Radiometer (AVHRR)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Huricane Satellite (HURSAT)-Advanced Very High Resolution Radiometer (AVHRR) is used to extend the HURSAT data set such that appling the Objective Dvorak technique...

  4. NanoComposite Polymers for High Resolution Near Infrared Detectors

    Data.gov (United States)

    National Aeronautics and Space Administration — Develop nanocomposite materials with tuned refractive index in the near infra red spectral range as an index-matched immersion lens for high resolution infra-red...

  5. Methodology of high-resolution photography for mural condition database

    Science.gov (United States)

    Higuchi, R.; Suzuki, T.; Shibata, M.; Taniguchi, Y.

    2015-08-01

    Digital documentation is one of the most useful techniques to record the condition of cultural heritage. Recently, high-resolution images become increasingly useful because it is possible to show general views of mural paintings and also detailed mural conditions in a single image. As mural paintings are damaged by environmental stresses, it is necessary to record the details of painting condition on high-resolution base maps. Unfortunately, the cost of high-resolution photography and the difficulty of operating its instruments and software have commonly been an impediment for researchers and conservators. However, the recent development of graphic software makes its operation simpler and less expensive. In this paper, we suggest a new approach to make digital heritage inventories without special instruments, based on our recent our research project in Üzümlü church in Cappadocia, Turkey. This method enables us to achieve a high-resolution image database with low costs, short time, and limited human resources.

  6. High resolution integral holography using Fourier ptychographic approach.

    Science.gov (United States)

    Li, Zhaohui; Zhang, Jianqi; Wang, Xiaorui; Liu, Delian

    2014-12-29

    An innovative approach is proposed for calculating high resolution computer generated integral holograms by using the Fourier Ptychographic (FP) algorithm. The approach initializes a high resolution complex hologram with a random guess, and then stitches together low resolution multi-view images, synthesized from the elemental images captured by integral imaging (II), to recover the high resolution hologram through an iterative retrieval with FP constrains. This paper begins with an analysis of the principle of hologram synthesis from multi-projections, followed by an accurate determination of the constrains required in the Fourier ptychographic integral-holography (FPIH). Next, the procedure of the approach is described in detail. Finally, optical reconstructions are performed and the results are demonstrated. Theoretical analysis and experiments show that our proposed approach can reconstruct 3D scenes with high resolution.

  7. High-resolution MRI in detecting subareolar breast abscess.

    Science.gov (United States)

    Fu, Peifen; Kurihara, Yasuyuki; Kanemaki, Yoshihide; Okamoto, Kyoko; Nakajima, Yasuo; Fukuda, Mamoru; Maeda, Ichiro

    2007-06-01

    Because subareolar breast abscess has a high recurrence rate, a more effective imaging technique is needed to comprehensively visualize the lesions and guide surgery. We performed a high-resolution MRI technique using a microscopy coil to reveal the characteristics and extent of subareolar breast abscess. High-resolution MRI has potential diagnostic value in subareolar breast abscess. This technique can be used to guide surgery with the aim of reducing the recurrence rate.

  8. High-resolution esophageal pressure topography for esophageal motility disorders

    OpenAIRE

    Hashem Fakhre Yaseri; Gholamreza Hamsi; Tayeb Ramim

    2016-01-01

    Background: High-resolution manometer (HRM) of the esophagus has become the main diagnostic test in the evaluation of esophageal motility disorders. The development of high-resolution manometry catheters and software displays of manometry recordings in color-coded pressure plots have changed the diagnostic assessment of esophageal disease. The first step of the Chicago classification described abnormal esophagogastric junction deglutitive relaxation. The latest classification system, proposed...

  9. Volumetric expiratory high-resolution CT of the lung

    International Nuclear Information System (INIS)

    Nishino, Mizuki; Hatabu, Hiroto

    2004-01-01

    We developed a volumetric expiratory high-resolution CT (HRCT) protocol that provides combined inspiratory and expiratory volumetric imaging of the lung without increasing radiation exposure, and conducted a preliminary feasibility assessment of this protocol to evaluate diffuse lung disease with small airway abnormalities. The volumetric expiratory high-resolution CT increased the detectability of the conducting airway to the areas of air trapping (P<0.0001), and added significant information about extent and distribution of air trapping (P<0.0001)

  10. Developing Visual Editors for High-Resolution Haptic Patterns

    DEFF Research Database (Denmark)

    Cuartielles, David; Göransson, Andreas; Olsson, Tony

    2012-01-01

    In this article we give an overview of our iterative work in developing visual editors for creating high resolution haptic patterns to be used in wearable, haptic feedback devices. During the past four years we have found the need to address the question of how to represent, construct and edit high...... resolution haptic patterns so that they translate naturally to the user’s haptic experience. To solve this question we have developed and tested several visual editors...

  11. EMODnet High Resolution Seabed Mapping - further developing a high resolution digital bathymetry for European seas

    Science.gov (United States)

    Schaap, D.; Schmitt, T.

    2017-12-01

    Access to marine data is a key issue for the EU Marine Strategy Framework Directive and the EU Marine Knowledge 2020 agenda and includes the European Marine Observation and Data Network (EMODnet) initiative. EMODnet aims at assembling European marine data, data products and metadata from diverse sources in a uniform way. The EMODnet Bathymetry project has developed Digital Terrain Models (DTM) for the European seas. These have been produced from survey and aggregated data sets that are indexed with metadata by adopting the SeaDataNet Catalogue services. SeaDataNet is a network of major oceanographic data centres around the European seas that manage, operate and further develop a pan-European infrastructure for marine and ocean data management. The latest EMODnet Bathymetry DTM release has a grid resolution of 1/8 arcminute and covers all European sea regions. Use has been made of circa 7800 gathered survey datasets and composite DTMs. Catalogues and the EMODnet DTM are published at the dedicated EMODnet Bathymetry portal including a versatile DTM viewing and downloading service. End December 2016 the Bathymetry project has been succeeded by EMODnet High Resolution Seabed Mapping (HRSM). This continues gathering of bathymetric in-situ data sets with extra efforts for near coastal waters and coastal zones. In addition Satellite Derived Bathymetry data are included to fill gaps in coverage of the coastal zones. The extra data and composite DTMs will increase the coverage of the European seas and its coastlines, and provide input for producing an EMODnet DTM with a common resolution of 1/16 arc minutes. The Bathymetry Viewing and Download service will be upgraded to provide a multi-resolution map and including 3D viewing. The higher resolution DTMs will also be used to determine best-estimates of the European coastline for a range of tidal levels (HAT, MHW, MSL, Chart Datum, LAT), thereby making use of a tidal model for Europe. Extra challenges will be `moving to the

  12. A method for generating high resolution satellite image time series

    Science.gov (United States)

    Guo, Tao

    2014-10-01

    There is an increasing demand for satellite remote sensing data with both high spatial and temporal resolution in many applications. But it still is a challenge to simultaneously improve spatial resolution and temporal frequency due to the technical limits of current satellite observation systems. To this end, much R&D efforts have been ongoing for years and lead to some successes roughly in two aspects, one includes super resolution, pan-sharpen etc. methods which can effectively enhance the spatial resolution and generate good visual effects, but hardly preserve spectral signatures and result in inadequate analytical value, on the other hand, time interpolation is a straight forward method to increase temporal frequency, however it increase little informative contents in fact. In this paper we presented a novel method to simulate high resolution time series data by combing low resolution time series data and a very small number of high resolution data only. Our method starts with a pair of high and low resolution data set, and then a spatial registration is done by introducing LDA model to map high and low resolution pixels correspondingly. Afterwards, temporal change information is captured through a comparison of low resolution time series data, and then projected onto the high resolution data plane and assigned to each high resolution pixel according to the predefined temporal change patterns of each type of ground objects. Finally the simulated high resolution data is generated. A preliminary experiment shows that our method can simulate a high resolution data with a reasonable accuracy. The contribution of our method is to enable timely monitoring of temporal changes through analysis of time sequence of low resolution images only, and usage of costly high resolution data can be reduces as much as possible, and it presents a highly effective way to build up an economically operational monitoring solution for agriculture, forest, land use investigation

  13. High resolution melting (HRM) analysis in sugar beet: identification of SNP markers associated to Fusarium resistance

    Science.gov (United States)

    Fusarium spp. cause severe damage in many agricultural crops including sugar beet. Sugar beet needs to be protected from these soil borne pathogens to guarantee an optimal sugar yield in the field. The genetic control is the key to overcoming this disease. Identification of single nucleotide polymor...

  14. Genetic high throughput screening in Retinitis Pigmentosa based on high resolution melting (HRM) analysis.

    Science.gov (United States)

    Anasagasti, Ander; Barandika, Olatz; Irigoyen, Cristina; Benitez, Bruno A; Cooper, Breanna; Cruchaga, Carlos; López de Munain, Adolfo; Ruiz-Ederra, Javier

    2013-11-01

    Retinitis Pigmentosa (RP) involves a group of genetically determined retinal diseases caused by a large number of mutations that result in rod photoreceptor cell death followed by gradual death of cone cells. Most cases of RP are monogenic, with more than 80 associated genes identified so far. The high number of genes and variants involved in RP, among other factors, is making the molecular characterization of RP a real challenge for many patients. Although HRM has been used for the analysis of isolated variants or single RP genes, as far as we are concerned, this is the first study that uses HRM analysis for a high-throughput screening of several RP genes. Our main goal was to test the suitability of HRM analysis as a genetic screening technique in RP, and to compare its performance with two of the most widely used NGS platforms, Illumina and PGM-Ion Torrent technologies. RP patients (n = 96) were clinically diagnosed at the Ophthalmology Department of Donostia University Hospital, Spain. We analyzed a total of 16 RP genes that meet the following inclusion criteria: 1) size: genes with transcripts of less than 4 kb; 2) number of exons: genes with up to 22 exons; and 3) prevalence: genes reported to account for, at least, 0.4% of total RP cases worldwide. For comparison purposes, RHO gene was also sequenced with Illumina (GAII; Illumina), Ion semiconductor technologies (PGM; Life Technologies) and Sanger sequencing (ABI 3130xl platform; Applied Biosystems). Detected variants were confirmed in all cases by Sanger sequencing and tested for co-segregation in the family of affected probands. We identified a total of 65 genetic variants, 15 of which (23%) were novel, in 49 out of 96 patients. Among them, 14 (4 novel) are probable disease-causing genetic variants in 7 RP genes, affecting 15 patients. Our HRM analysis-based study, proved to be a cost-effective and rapid method that provides an accurate identification of genetic RP variants. This approach is effective for medium sized (<4 kb transcript) RP genes, which constitute over 80% of the total of known RP genes.

  15. High resolution melting detects sequence polymorphism in rubus occidentalis L. monomorphic microsatellite markers

    Science.gov (United States)

    Microsatellite, or simple sequence repeat (SSR) markers, are valuable as co-dominant genetic markers with a variety of applications such as DNA fingerprinting, linkage mapping, and population structure analysis. However, primer pairs designed from the regions that flank SSRs often generate fragment...

  16. Detection of Sequence Polymorphism in Rubus Occidentalis L. Monomorphic Microsatellite Markers by High Resolution Melting

    Science.gov (United States)

    Microsatellite, or simple sequence repeat (SSR) markers, are valuable as co-dominant genetic markers with a variety of applications such as DNA fingerprinting, linkage mapping, and population structure analysis. Development of microsatellite primers through the identification of appropriate repeate...

  17. Identification of rare and frequent variants of the CASR gene by high-resolution melting

    DEFF Research Database (Denmark)

    Nissen, Peter H; Christensen, Signe E; Ladefoged, Søren A

    2012-01-01

    of seven new CASR variants and nine recurrent. HRM variant scanning, in combination with small amplicon genotyping, provides a simple workflow with reduced sequencing burden. Bioinformatics analyses using two freely available prediction tools (PolyPhen2 and SIFT) for evaluating amino acid substitutions...

  18. High resolution manometry findings in patients with esophageal epiphrenic diverticula.

    Science.gov (United States)

    Vicentine, Fernando P P; Herbella, Fernando A M; Silva, Luciana C; Patti, Marco G

    2011-12-01

    The pathophysiology of esophageal epiphrenic diverticula is still uncertain even though a concomitant motility disorder is found in the majority of patients in different series. High resolution manometry may allow detection of motor abnormalities in a higher number of patients with esophageal epiphrenic diverticula compared with conventional manometry. This study aims to evaluate the high resolution manometry findings in patients with esophageal epiphrenic diverticula. Nine individuals (mean age 63 ± 10 years, 4 females) with esophageal epiphrenic diverticula underwent high resolution manometry. A single diverticulum was observed in eight patients and multiple diverticula in one. Visual analysis of conventional tracings and color pressure plots for identification of segmental abnormalities was performed by two researchers experienced in high resolution manometry. Upper esophageal sphincter was normal in all patients. Esophageal body was abnormal in eight patients; lower esophageal sphincter was abnormal in seven patients. Named esophageal motility disorders were found in seven patients: achalasia in six, diffuse esophageal spasm in one. In one patient, a segmental hypercontractile zone was noticed with pressure of 196 mm Hg. High resolution manometry demonstrated motor abnormalities in all patients with esophageal epiphrenic diverticula.

  19. Compact and high-resolution optical orbital angular momentum sorter

    Directory of Open Access Journals (Sweden)

    Chenhao Wan

    2017-03-01

    Full Text Available A compact and high-resolution optical orbital angular momentum (OAM sorter is proposed and demonstrated. The sorter comprises a quadratic fan-out mapper and a dual-phase corrector positioned in the pupil plane and the Fourier plane, respectively. The optical system is greatly simplified compared to previous demonstrations of OAM sorting, and the performance in resolution and efficiency is maintained. A folded configuration is set up using a single reflective spatial light modulator (SLM to demonstrate the validity of the scheme. The two phase elements are implemented on the left and right halves of the SLM and connected by a right-angle prism. Experimental results demonstrate the high resolution of the compact OAM sorter, and the current limit in efficiency can be overcome by replacing with transmissive SLMs and removing the beam splitters. This novel scheme paves the way for the miniaturization and integration of high-resolution OAM sorters.

  20. High-resolution SPECT for small-animal imaging

    International Nuclear Information System (INIS)

    Qi Yujin

    2006-01-01

    This article presents a brief overview of the development of high-resolution SPECT for small-animal imaging. A pinhole collimator has been used for high-resolution animal SPECT to provide better spatial resolution and detection efficiency in comparison with a parallel-hole collimator. The theory of imaging characteristics of the pinhole collimator is presented and the designs of the pinhole aperture are discussed. The detector technologies used for the development of small-animal SPECT and the recent advances are presented. The evolving trend of small-animal SPECT is toward a multi-pinhole and a multi-detector system to obtain a high resolution and also a high detection efficiency. (authors)

  1. High-resolution spectroscopy of gases for industrial applications

    DEFF Research Database (Denmark)

    Fateev, Alexander; Clausen, Sønnik

    High-resolution spectroscopy of gases is a powerful technique which has various fundamental and practical applications: in situ simultaneous measurements of gas temperature and gas composition, radiative transfer modeling, validation of existing and developing of new databases and etc. Existing...... databases (e.g. HITRAN, HITEMP or CDSD) can normally be used for absorption spectra calculations at limited temperature/pressure ranges. Therefore experimental measurements of absorption/transmission spectra gases (e.g. CO2, H2O or SO2) at high-resolution and elevated temperatures are essential both...... for analysis of complex experimental data and further development of the databases. High-temperature gas cell facilities available at DTU Chemical Engineering are presented and described. The gas cells and high-resolution spectrometers allow us to perform high-quality reference measurements of gases relevant...

  2. 1024 matrix image reconstruction: usefulness in high resolution chest CT

    International Nuclear Information System (INIS)

    Jeong, Sun Young; Chung, Myung Jin; Chong, Se Min; Sung, Yon Mi; Lee, Kyung Soo

    2006-01-01

    We tried to evaluate whether high resolution chest CT with a 1,024 matrix has a significant advantage in image quality compared to a 512 matrix. Each set of 512 and 1024 matrix high resolution chest CT scans with both 0.625 mm and 1.25 mm slice thickness were obtained from 26 patients. Seventy locations that contained twenty-four low density lesions without sharp boundary such as emphysema, and forty-six sharp linear densities such as linear fibrosis were selected; these were randomly displayed on a five mega pixel LCD monitor. All the images were masked for information concerning the matrix size and slice thickness. Two chest radiologists scored the image quality of each ar rowed lesion as follows: (1) undistinguishable, (2) poorly distinguishable, (3) fairly distinguishable, (4) well visible and (5) excellently visible. The scores were compared from the aspects of matrix size, slice thickness and the different observers by using ANOVA tests. The average and standard deviation of image quality were 3.09 (± .92) for the 0.625 mm x 512 matrix, 3.16 (± .84) for the 0.625 mm x 1024 matrix, 2.49 (± 1.02) for the 1.25 mm x 512 matrix, and 2.35 (± 1.02) for the 1.25 mm x 1024 matrix, respectively. The image quality on both matrices of the high resolution chest CT scans with a 0.625 mm slice thickness was significantly better than that on the 1.25 mm slice thickness (ρ < 0.001). However, the image quality on the 1024 matrix high resolution chest CT scans was not significantly different from that on the 512 matrix high resolution chest CT scans (ρ = 0.678). The interobserver variation between the two observers was not significant (ρ = 0.691). We think that 1024 matrix image reconstruction for high resolution chest CT may not be clinical useful

  3. Application of the specific thermal properties of Ag nanoparticles to high-resolution metal patterning

    International Nuclear Information System (INIS)

    Son, Yong; Yeo, Junyeob; Ha, Cheol Woo; Lee, Jinhwan; Hong, Sukjoon; Nam, Koo Hyun; Yang, Dong-Yol; Ko, Seung Hwan

    2012-01-01

    Metal nanoparticles exhibit specific electronic, chemical and optical properties due to the thermodynamic size effect, which cannot be observed in bulk materials. Ag NPs show size dependent melting temperature depression phenomena. In this study, the thermal sintering behavior of the self-assembled monolayer protected Ag NPs has been observed using in situ transmission electron microscopy. The thermal characteristics of the Ag NPs have also been examined with a thermogravimetric analysis, a differential scanning calorimetry and a thermal conductivity measurement. These assessments have shown that the melting of the Ag NPs starts at 150 °C, which is much lower than the melting temperature of bulk silver (960 °C). The measured thermal conductivity of the Ag NPs (0.37 W/(m K)) is also lower than that of bulk silver (429 W/(m K)). These specific thermal properties of the Ag NPs can be applied to a low-temperature and a high-resolution direct-metal patterning process.

  4. Reproducible high-resolution multispectral image acquisition in dermatology

    Science.gov (United States)

    Duliu, Alexandru; Gardiazabal, José; Lasser, Tobias; Navab, Nassir

    2015-07-01

    Multispectral image acquisitions are increasingly popular in dermatology, due to their improved spectral resolution which enables better tissue discrimination. Most applications however focus on restricted regions of interest, imaging only small lesions. In this work we present and discuss an imaging framework for high-resolution multispectral imaging on large regions of interest.

  5. Dye laser light for high-resolution classical photography

    International Nuclear Information System (INIS)

    Geissler, K.K.

    1982-01-01

    The test run with the bubble chamber HOLEBC in October 1981 offered the opportunity of checking the usefulness of de-speckled dye laser light for illumination purposes in high-resolution classical dark field photography of small bubble chambers. (orig./HSI)

  6. High-resolution seismic imaging of the Sohagpur Gondwana basin ...

    Indian Academy of Sciences (India)

    The quality of the high-resolution seismic data depends mainly on the data ..... metric rift geometry. Based on the .... Biswas S K 2003 Regional tectonic framework of the .... Sheth H C, Ray J S, Ray R, Vanderkluysen L, Mahoney J. J, Kumar A ...

  7. Pulmonary Gaucher's disease: high-resolution computed tomographic features

    International Nuclear Information System (INIS)

    Tunaci, A.; Berkmen, Y.M.; Goekmen, E.

    1995-01-01

    CT findings in pulmonary Gaucher's disease have not been previously reported. Chest radiograph of a patient with pulmonary involvement in type I Gaucher's disease proven by biopsy showed linear and reticulo-nodular opacities. High-resolution CT demonstrated thickening of the interlobular septa and between four and six small nodules within secondary lobules, probably each corresponding to an acinus. (orig.)

  8. High resolution techniques using scanning proton microprobe (SPM)

    International Nuclear Information System (INIS)

    Cholewa, M.; Saint, A.; Prawer, S.; Laird, J.S.; Legge, G.J.F.; Bardos, R.A.; Moorhead, G.F.; Taylor, G.N.; Stuart, S.A.; Howard, J.

    1994-01-01

    The very high resolution (down to 50 nm) achieved with low beam currents (fA) in a scanning ion microprobe have lead to many nondestructive techniques of microanalysis. This paper discusses recent developments and applications in the use of 3-D STIM (scanning transmission ion microscopy) Tomography, channeling STIM and IBIC (ion beam induced charge). (orig.)

  9. Structure Identification in High-Resolution Transmission Electron Microscopic Images

    DEFF Research Database (Denmark)

    Vestergaard, Jacob Schack; Kling, Jens; Dahl, Anders Bjorholm

    2014-01-01

    A connection between microscopic structure and macroscopic properties is expected for almost all material systems. High-resolution transmission electron microscopy is a technique offering insight into the atomic structure, but the analysis of large image series can be time consuming. The present ...

  10. Role of land state in a high resolution mesoscale model

    Indian Academy of Sciences (India)

    ... Proceedings – Mathematical Sciences · Resonance – Journal of Science ... Land surface characteristics; high resolution mesoscale model; Uttarakhand ... to predict realistic location, timing, amount,intensity and distribution of rainfall ... region embedded within two low pressure centers over Arabian Seaand Bay of Bengal.

  11. A high resolution powder diffractometer using focusing optics

    Indian Academy of Sciences (India)

    E-mail: siruguri@csr.ernet.in. Abstract. In this paper, we describe the design, construction and performance of a new high resolution neutron powder diffractometer that has been installed at the Dhruva reactor, Trombay, India. The instrument employs novel design concepts like the use of bent, perfect crystal monochromator ...

  12. Application of high resolution SNP arrays in patients with congenital ...

    Indian Academy of Sciences (India)

    clinical experience in implementing whole-genome high-resolution SNP arrays to investigate 33 patients with syndromic and .... Online Mendelian Inheritance in Man database (OMIM, ..... of damaged mitochondria through either autophagy or mito- ..... malformations: associations with maternal and infant character- istics in a ...

  13. Workshop on high-resolution, large-acceptance spectrometers

    Energy Technology Data Exchange (ETDEWEB)

    Zeidman, B. (ed.)

    1981-01-01

    The purpose of the Workshop on High-Resolution, Large-Acceptance Spectrometers was to provide a means for exchange of information among those actively engaged in the design and construction of these new spectrometers. Thirty-seven papers were prepared for the data base.

  14. Yeast expression proteomics by high-resolution mass spectrometry

    DEFF Research Database (Denmark)

    Walther, Tobias C; Olsen, Jesper Velgaard; Mann, Matthias

    2010-01-01

    -translational controls contribute majorly to regulation of protein abundance, for example in heat shock stress response. The development of new sample preparation methods, high-resolution mass spectrometry and novel bioinfomatic tools close this gap and allow the global quantitation of the yeast proteome under different...

  15. High resolution satellite imagery : from spies to pipeline management

    Energy Technology Data Exchange (ETDEWEB)

    Adam, S. [Canadian Geomatic Solutions Ltd., Calgary, AB (Canada); Farrell, M. [TransCanada Transmission, Calgary, AB (Canada)

    2000-07-01

    The launch of Space Imaging's IKONOS satellite in September 1999 has opened the door for corridor applications. The technology has been successfully implemented by TransCanada PipeLines in mapping over 1500 km of their mainline. IKONOS is the world's first commercial high resolution satellite which collects data at 1-meter black/white and 4-meter multi-spectral. Its use is regulated by the U.S. government. It is the best source of high resolution satellite image data. Other sources include the Indian Space Agency's IRS-1 C/D satellite and the Russian SPIN-2 which provides less reliable coverage. In addition, two more high resolution satellites may be launched this year to provide imagery every day of the year. IKONOS scenes as narrow as 5 km can be purchased. TransCanada conducted a pilot study to determine if high resolution satellite imagery is as effective as ortho-photos for identifying population structures within a buffer of TransCanada's east line right-of-way. The study examined three unique segments where residential, commercial, industrial and public features were compared. It was determined that IKONOS imagery is as good as digital ortho-photos for updating structures from low to very high density areas. The satellite imagery was also logistically easier than ortho-photos to acquire. This will be even more evident when the IKONOS image archives begins to grow. 4 tabs., 3 figs.

  16. Novel techniques in VUV high-resolution spectroscopy

    NARCIS (Netherlands)

    Ubachs, W.M.G.; Salumbides, E.J.; Eikema, K.S.E.; de Oliveira, N.; Nahon, L.

    2014-01-01

    Novel VUV sources and techniques for VUV spectroscopy are reviewed. Laser-based VUV sources have been developed via non-linear upconversion of laser pulses in the nanosecond (ns), the picosecond (ps), and femtosecond (fs) domain, and are applied in high-resolution gas phase spectroscopic studies.

  17. High resolution X-ray diffraction studies on unirradiated

    Indian Academy of Sciences (India)

    High-resolution X-ray diffraction technique, employing a three-crystal monochromator–collimator combination is used to study the irradiation induced defects in flux grown Sr-hexaferrite crystals irradiated with 50 MeV Li3+ ion beams at room temperature with a fluence value of 1 × 1014 ions/cm2. The diffraction curves of the ...

  18. High resolution STEM of quantum dots and quantum wires

    DEFF Research Database (Denmark)

    Kadkhodazadeh, Shima

    2013-01-01

    This article reviews the application of high resolution scanning transmission electron microscopy (STEM) to semiconductor quantum dots (QDs) and quantum wires (QWRs). Different imaging and analytical techniques in STEM are introduced and key examples of their application to QDs and QWRs...

  19. Pattern of interstitial lung disease detected by high resolution ...

    African Journals Online (AJOL)

    Background: Diffuse lung diseases constitute a major cause of morbidity and mortality worldwide. High Resolution Computed Tomography (HRCT) is the recommended imaging technique in the diagnosis, assessment and followup of these diseases. Objectives: To describe the pattern of HRCT findings in patients with ...

  20. A multi-channel high-resolution time recorder system

    International Nuclear Information System (INIS)

    Zhang Lingyun; Yang Xiaojun; Song Kezhu; Wang Yanfang

    2004-01-01

    This paper introduces a multi-channel and high-speed time recorder system, which was originally designed to work in the experiments of quantum cryptography research. The novelty of the system is that all the hardware logic is performed by only one FPGA. The system can achieve several desirable features, such as simplicity, high resolution and high processing speed. (authors)

  1. Remote parallel rendering for high-resolution tiled display walls

    KAUST Repository

    Nachbaur, Daniel

    2014-11-01

    © 2014 IEEE. We present a complete, robust and simple to use hardware and software stack delivering remote parallel rendering of complex geometrical and volumetric models to high resolution tiled display walls in a production environment. We describe the setup and configuration, present preliminary benchmarks showing interactive framerates, and describe our contributions for a seamless integration of all the software components.

  2. Remote parallel rendering for high-resolution tiled display walls

    KAUST Repository

    Nachbaur, Daniel; Dumusc, Raphael; Bilgili, Ahmet; Hernando, Juan; Eilemann, Stefan

    2014-01-01

    © 2014 IEEE. We present a complete, robust and simple to use hardware and software stack delivering remote parallel rendering of complex geometrical and volumetric models to high resolution tiled display walls in a production environment. We describe the setup and configuration, present preliminary benchmarks showing interactive framerates, and describe our contributions for a seamless integration of all the software components.

  3. Human enamel structure studied by high resolution electron microscopy

    International Nuclear Information System (INIS)

    Wen, S.L.

    1989-01-01

    Human enamel structural features are characterized by high resolution electron microscopy. The human enamel consists of polycrystals with a structure similar to Ca10(PO4)6(OH)2. This article describes the structural features of human enamel crystal at atomic and nanometer level. Besides the structural description, a great number of high resolution images are included. Research into the carious process in human enamel is very important for human beings. This article firstly describes the initiation of caries in enamel crystal at atomic and unit-cell level and secondly describes the further steps of caries with structural and chemical demineralization. The demineralization in fact, is the origin of caries in human enamel. The remineralization of carious areas in human enamel has drawn more and more attention as its potential application is realized. This process has been revealed by high resolution electron microscopy in detail in this article. On the other hand, the radiation effects on the structure of human enamel are also characterized by high resolution electron microscopy. In order to reveal this phenomenon clearly, a great number of electron micrographs have been shown, and a physical mechanism is proposed. 26 references

  4. High resolution and high speed positron emission tomography data acquisition

    International Nuclear Information System (INIS)

    Burgiss, S.G.; Byars, L.G.; Jones, W.F.; Casey, M.E.

    1986-01-01

    High resolution positron emission tomography (PET) requires many detectors. Thus, data collection systems for PET must have high data rates, wide data paths, and large memories to histogram the events. This design uses the VMEbus to cost effectively provide these features. It provides for several modes of operation including real time sorting, list mode data storage, and replay of stored list mode data

  5. High resolution wind turbine wake measurements with a scanning lidar

    DEFF Research Database (Denmark)

    Herges, T. G.; Maniaci, D. C.; Naughton, B. T.

    2017-01-01

    High-resolution lidar wake measurements are part of an ongoing field campaign being conducted at the Scaled Wind Farm Technology facility by Sandia National Laboratories and the National Renewable Energy Laboratory using a customized scanning lidar from the Technical University of Denmark. One...

  6. High resolution mid-infrared spectroscopy based on frequency upconversion

    DEFF Research Database (Denmark)

    Dam, Jeppe Seidelin; Hu, Qi; Tidemand-Lichtenberg, Peter

    2013-01-01

    signals can be analyzed. The obtainable frequency resolution is usually in the nm range where sub nm resolution is preferred in many applications, like gas spectroscopy. In this work we demonstrate how to obtain sub nm resolution when using upconversion. In the presented realization one object point...... high resolution spectral performance by observing emission from hot water vapor in a butane gas burner....

  7. Systematic high-resolution assessment of global hydropower potential

    NARCIS (Netherlands)

    Hoes, Olivier A C; Meijer, Lourens J J; Van Der Ent, Ruud J.|info:eu-repo/dai/nl/364164794; Van De Giesen, Nick C.

    2017-01-01

    Population growth, increasing energy demand and the depletion of fossil fuel reserves necessitate a search for sustainable alternatives for electricity generation. Hydropower could replace a large part of the contribution of gas and oil to the present energy mix. However, previous high-resolution

  8. High-Resolution Geologic Mapping of Martian Terraced Fan Deposits

    Science.gov (United States)

    Wolak, J. M.; Patterson, A. B.; Smith, S. D.; Robbins, N. N.

    2018-06-01

    This abstract documents our initial progress (year 1) mapping terraced fan features on Mars. Our objective is to investigate the role of fluids during fan formation and produce the first high-resolution geologic map (1:18k) of a terraced fan.

  9. High-resolution axial MR imaging of tibial stress injuries

    Directory of Open Access Journals (Sweden)

    Mammoto Takeo

    2012-05-01

    Full Text Available Abstract Purpose To evaluate the relative involvement of tibial stress injuries using high-resolution axial MR imaging and the correlation with MR and radiographic images. Methods A total of 33 patients with exercise-induced tibial pain were evaluated. All patients underwent radiograph and high-resolution axial MR imaging. Radiographs were taken at initial presentation and 4 weeks later. High-resolution MR axial images were obtained using a microscopy surface coil with 60 × 60 mm field of view on a 1.5T MR unit. All images were evaluated for abnormal signals of the periosteum, cortex and bone marrow. Results Nineteen patients showed no periosteal reaction at initial and follow-up radiographs. MR imaging showed abnormal signals in the periosteal tissue and partially abnormal signals in the bone marrow. In 7 patients, periosteal reaction was not seen at initial radiograph, but was detected at follow-up radiograph. MR imaging showed abnormal signals in the periosteal tissue and entire bone marrow. Abnormal signals in the cortex were found in 6 patients. The remaining 7 showed periosteal reactions at initial radiograph. MR imaging showed abnormal signals in the periosteal tissue in 6 patients. Abnormal signals were seen in the partial and entire bone marrow in 4 and 3 patients, respectively. Conclusions Bone marrow abnormalities in high-resolution axial MR imaging were related to periosteal reactions at follow-up radiograph. Bone marrow abnormalities might predict later periosteal reactions, suggesting shin splints or stress fractures. High-resolution axial MR imaging is useful in early discrimination of tibial stress injuries.

  10. High-resolution axial MR imaging of tibial stress injuries

    Science.gov (United States)

    2012-01-01

    Purpose To evaluate the relative involvement of tibial stress injuries using high-resolution axial MR imaging and the correlation with MR and radiographic images. Methods A total of 33 patients with exercise-induced tibial pain were evaluated. All patients underwent radiograph and high-resolution axial MR imaging. Radiographs were taken at initial presentation and 4 weeks later. High-resolution MR axial images were obtained using a microscopy surface coil with 60 × 60 mm field of view on a 1.5T MR unit. All images were evaluated for abnormal signals of the periosteum, cortex and bone marrow. Results Nineteen patients showed no periosteal reaction at initial and follow-up radiographs. MR imaging showed abnormal signals in the periosteal tissue and partially abnormal signals in the bone marrow. In 7 patients, periosteal reaction was not seen at initial radiograph, but was detected at follow-up radiograph. MR imaging showed abnormal signals in the periosteal tissue and entire bone marrow. Abnormal signals in the cortex were found in 6 patients. The remaining 7 showed periosteal reactions at initial radiograph. MR imaging showed abnormal signals in the periosteal tissue in 6 patients. Abnormal signals were seen in the partial and entire bone marrow in 4 and 3 patients, respectively. Conclusions Bone marrow abnormalities in high-resolution axial MR imaging were related to periosteal reactions at follow-up radiograph. Bone marrow abnormalities might predict later periosteal reactions, suggesting shin splints or stress fractures. High-resolution axial MR imaging is useful in early discrimination of tibial stress injuries. PMID:22574840

  11. Analysis of Chinese women with primary ovarian insufficiency by high resolution array-comparative genomic hybridization.

    Science.gov (United States)

    Liao, Can; Fu, Fang; Yang, Xin; Sun, Yi-Min; Li, Dong-Zhi

    2011-06-01

    Primary ovarian insufficiency (POI) is defined as a primary ovarian defect characterized by absent menarche (primary amenorrhea) or premature depletion of ovarian follicles before the age of 40 years. The etiology of primary ovarian insufficiency in human female patients is still unclear. The purpose of this study is to investigate the potential genetic causes in primary amenorrhea patients by high resolution array based comparative genomic hybridization (array-CGH) analysis. Following the standard karyotyping analysis, genomic DNA from whole blood of 15 primary amenorrhea patients and 15 normal control women was hybridized with Affymetrix cytogenetic 2.7M arrays following the standard protocol. Copy number variations identified by array-CGH were confirmed by real time polymerase chain reaction. All the 30 samples were negative by conventional karyotyping analysis. Microdeletions on chromosome 17q21.31-q21.32 with approximately 1.3 Mb were identified in four patients by high resolution array-CGH analysis. This included the female reproductive secretory pathway related factor N-ethylmaleimide-sensitive factor (NSF) gene. The results of the present study suggest that there may be critical regions regulating primary ovarian insufficiency in women with a 17q21.31-q21.32 microdeletion. This effect might be due to the loss of function of the NSF gene/genes within the deleted region or to effects on contiguous genes.

  12. High-resolution transmission electron microscopy of grain-refining particles in amorphous aluminum alloys

    International Nuclear Information System (INIS)

    Schumacher, P.; Greer, A.L.

    1996-01-01

    The nucleation mechanism of Al-Ti-B grain refiners is studied in an Al-based amorphous alloy. The ability to limit growth of α-Al in the amorphous alloy permits the microscopical observation of nucleation events on boride particles. Earlier studies of this kind are extended by using high-resolution electron microscopy. This shows that the efficient nucleation α-Al depends on the TiB 2 particles being coated with a thin layer of Al 3 Ti, which can form only when there is some excess titanium in the melt. The aluminide layer, stabilized by adsorption effects, can be as little as a few monolayers thick, and is coherent with the boride. The nature of this layer, and its importance for the nucleation mechanism are discussed. The fading of the grain refinement action is also considered

  13. High resolution selective multilayer laser processing by nanosecond laser ablation of metal nanoparticle films

    International Nuclear Information System (INIS)

    Ko, Seung H.; Pan Heng; Hwang, David J.; Chung, Jaewon; Ryu, Sangil; Grigoropoulos, Costas P.; Poulikakos, Dimos

    2007-01-01

    Ablation of gold nanoparticle films on polymer was explored using a nanosecond pulsed laser, with the goal to achieve feature size reduction and functionality not amenable with inkjet printing. The ablation threshold fluence for the unsintered nanoparticle deposit was at least ten times lower than the reported threshold for the bulk film. This could be explained by the combined effects of melting temperature depression, lower conductive heat transfer loss, strong absorption of the incident laser beam, and the relatively weak bonding between nanoparticles. The ablation physics were verified by the nanoparticle sintering characterization, ablation threshold measurement, time resolved ablation plume shadowgraphs, analysis of ablation ejecta, and the measurement and calculation of optical properties. High resolution and clean feature fabrication with small energy and selective multilayer processing are demonstrated

  14. High-resolution x-ray imaging using a structured scintillator

    Energy Technology Data Exchange (ETDEWEB)

    Hormozan, Yashar, E-mail: hormozan@kth.se; Sychugov, Ilya; Linnros, Jan [Materials and Nano Physics, School of Information and Communication Technology, KTH Royal Institute of Technology, Electrum 229, Kista, Stockholm SE-16440 (Sweden)

    2016-02-15

    Purpose: In this study, the authors introduce a new generation of finely structured scintillators with a very high spatial resolution (a few micrometers) compared to conventional scintillators, yet maintaining a thick absorbing layer for improved detectivity. Methods: Their concept is based on a 2D array of high aspect ratio pores which are fabricated by ICP etching, with spacings (pitches) of a few micrometers, on silicon and oxidation of the pore walls. The pores were subsequently filled by melting of powdered CsI(Tl), as the scintillating agent. In order to couple the secondary emitted photons of the back of the scintillator array to a CCD device, having a larger pixel size than the pore pitch, an open optical microscope with adjustable magnification was designed and implemented. By imaging a sharp edge, the authors were able to calculate the modulation transfer function (MTF) of this finely structured scintillator. Results: The x-ray images of individually resolved pores suggest that they have been almost uniformly filled, and the MTF measurements show the feasibility of a few microns spatial resolution imaging, as set by the scintillator pore size. Compared to existing techniques utilizing CsI needles as a structured scintillator, their results imply an almost sevenfold improvement in resolution. Finally, high resolution images, taken by their detector, are presented. Conclusions: The presented work successfully shows the functionality of their detector concept for high resolution imaging and further fabrication developments are most likely to result in higher quantum efficiencies.

  15. High resolution color imagery for orthomaps and remote sensing

    Energy Technology Data Exchange (ETDEWEB)

    Fricker, Peter [Leica Geosystems GIS and Mapping, LLC (Switzerland); Gallo, M. Guillermo [Leica Geosystems GIS and Mapping, LLC (United States)

    2005-07-01

    The ADS40 Airborne Digital Pushbroom Sensor is currently the only commercial sensor capable of acquiring color and false color strip images in the low decimeter range at the same high resolution as the black and white stereo images. This high resolution of 12,000 pixels across the entire swath and 100% forward overlap in the image strips result in high quality DSM's, True Ortho's and at the same time allow unbiased remote sensing applications due to color strip images unchanged by pan-sharpening. The paper gives details on how the pushbroom sensor achieves these seemingly difficult technical challenges. It describes how a variety of mapping applications benefit from this sensor, a sensor which acts as a satellite pushbroom sensor within the airborne environment. (author)

  16. Ring artifact correction for high-resolution micro CT

    International Nuclear Information System (INIS)

    Kyriakou, Yiannis; Prell, Daniel; Kalender, Willi A

    2009-01-01

    In high-resolution micro CT using flat detectors (FD), imperfect or defect detector elements may cause concentric-ring artifacts due to their continuous over- or underestimation of attenuation values, which often disturb image quality. We here present a dedicated image-based ring artifact correction method for high-resolution micro CT, based on median filtering of the reconstructed image and working on a transformed version of the reconstructed images in polar coordinates. This post-processing method reduced ring artifacts in the reconstructed images and improved image quality for phantom and in in vivo scans. Noise and artifacts were reduced both in transversal and in multi-planar reformations along the longitudinal axis. (note)

  17. High resolution spectroscopy in solids by nuclear magnetic resonance

    International Nuclear Information System (INIS)

    Bonagamba, T.J.

    1991-07-01

    The nuclear magnetic resonance (NMR) techniques for High Resolution Spectroscopy in Solids are described. Also the construction project of a partially home made spectrometer and its applications in the characterization of solid samples are shown in detail. The high resolution spectrometer used is implemented with the double resonance multiple pulses sequences and magic angle spinning (MAS) and can be used with solid and liquid samples. The maximum spinning frequency for the MAS experiment is in excess of 5 Khz, the double resonance sequences can be performed with any type of nucleus, in the variable temperature operating range with nitrogen gas: -120 0 C to +160 0 C, and is fully controlled by a Macintosh IIci microcomputer. (author)

  18. High resolution radar satellite imagery analysis for safeguards applications

    Energy Technology Data Exchange (ETDEWEB)

    Minet, Christian; Eineder, Michael [German Aerospace Center, Remote Sensing Technology Institute, Department of SAR Signal Processing, Wessling, (Germany); Rezniczek, Arnold [UBA GmbH, Herzogenrath, (Germany); Niemeyer, Irmgard [Forschungszentrum Juelich, Institue of Energy and Climate Research, IEK-6: Nuclear Waste Management and Reactor Safety, Juelich, (Germany)

    2011-12-15

    For monitoring nuclear sites, the use of Synthetic Aperture Radar (SAR) imagery shows essential promises. Unlike optical remote sensing instruments, radar sensors operate under almost all weather conditions and independently of the sunlight, i.e. time of the day. Such technical specifications are required both for continuous and for ad-hoc, timed surveillance tasks. With Cosmo-Skymed, TerraSARX and Radarsat-2, high-resolution SAR imagery with a spatial resolution up to 1m has recently become available. Our work therefore aims to investigate the potential of high-resolution TerraSAR data for nuclear monitoring. This paper focuses on exploiting amplitude of a single acquisition, assessing amplitude changes and phase differences between two acquisitions, and PS-InSAR processing of an image stack.

  19. High resolution reservoir geological modelling using outcrop information

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Changmin; Lin Kexiang; Liu Huaibo [Jianghan Petroleum Institute, Hubei (China)] [and others

    1997-08-01

    This is China`s first case study of high resolution reservoir geological modelling using outcrop information. The key of the modelling process is to build a prototype model and using the model as a geological knowledge bank. Outcrop information used in geological modelling including seven aspects: (1) Determining the reservoir framework pattern by sedimentary depositional system and facies analysis; (2) Horizontal correlation based on the lower and higher stand duration of the paleo-lake level; (3) Determining the model`s direction based on the paleocurrent statistics; (4) Estimating the sandbody communication by photomosaic and profiles; (6) Estimating reservoir properties distribution within sandbody by lithofacies analysis; and (7) Building the reservoir model in sandbody scale by architectural element analysis and 3-D sampling. A high resolution reservoir geological model of Youshashan oil field has been built by using this method.

  20. High resolution spectroscopy in the microwave and far infrared

    Science.gov (United States)

    Pickett, Herbert M.

    1990-01-01

    High resolution rotational spectroscopy has long been central to remote sensing techniques in atmospheric sciences and astronomy. As such, laboratory measurements must supply the required data to make direct interpretation of data for instruments which sense atmospheres using rotational spectra. Spectral measurements in the microwave and far infrared regions are also very powerful tools when combined with infrared measurements for characterizing the rotational structure of vibrational spectra. In the past decade new techniques were developed which have pushed high resolution spectroscopy into the wavelength region between 25 micrometers and 2 mm. Techniques to be described include: (1) harmonic generation of microwave sources, (2) infrared laser difference frequency generation, (3) laser sideband generation, and (4) ultrahigh resolution interferometers.

  1. High-resolution observation by double-biprism electron holography

    International Nuclear Information System (INIS)

    Harada, Ken; Tonomura, Akira; Matsuda, Tsuyoshi; Akashi, Tetsuya; Togawa, Yoshihiko

    2004-01-01

    High-resolution electron holography has been achieved by using a double-biprism interferometer implemented on a 1 MV field emission electron microscope. The interferometer was installed behind the first magnifying lens to narrow carrier fringes and thus enabled complete separation of sideband Fourier spectrum from center band in reconstruction process. Holograms of Au fine particles and single-crystalline thin films with the finest fringe spacing of 4.2 pm were recorded and reconstructed. The overall holography system including the reconstruction process performed well for holograms in which carrier fringes had a spacing of around 10 pm. High-resolution lattice images of the amplitude and phase were clearly reconstructed without mixing of the center band and sideband information. Additionally, entire holograms were recorded without Fresnel fringes normally generated by the filament electrode of the biprism, and the holograms were thus reconstructed without the artifacts caused by Fresnel fringes

  2. Automated data processing of high-resolution mass spectra

    DEFF Research Database (Denmark)

    Hansen, Michael Adsetts Edberg; Smedsgaard, Jørn

    of the massive amounts of data. We present an automated data processing method to quantitatively compare large numbers of spectra from the analysis of complex mixtures, exploiting the full quality of high-resolution mass spectra. By projecting all detected ions - within defined intervals on both the time...... infusion of crude extracts into the source taking advantage of the high sensitivity, high mass resolution and accuracy and the limited fragmentation. Unfortunately, there has not been a comparable development in the data processing techniques to fully exploit gain in high resolution and accuracy...... infusion analyses of crude extract to find the relationship between species from several species terverticillate Penicillium, and also that the ions responsible for the segregation can be identified. Furthermore the process can automate the process of detecting unique species and unique metabolites....

  3. Towards high-resolution positron emission tomography for small volumes

    International Nuclear Information System (INIS)

    McKee, B.T.A.

    1982-01-01

    Some arguments are made regarding the medical usefulness of high spatial resolution in positron imaging, even if limited to small imaged volumes. Then the intrinsic limitations to spatial resolution in positron imaging are discussed. The project to build a small-volume, high resolution animal research prototype (SHARP) positron imaging system is described. The components of the system, particularly the detectors, are presented and brief mention is made of data acquisition and image reconstruction methods. Finally, some preliminary imaging results are presented; a pair of isolated point sources and 18 F in the bones of a rabbit. Although the detector system is not fully completed, these first results indicate that the goals of high sensitivity and high resolution (4 mm) have been realized. (Auth.)

  4. High-resolution investigations of edge effects in neutron imaging

    International Nuclear Information System (INIS)

    Strobl, M.; Kardjilov, N.; Hilger, A.; Kuehne, G.; Frei, G.; Manke, I.

    2009-01-01

    Edge enhancement is the main effect measured by the so-called inline or propagation-based neutron phase contrast imaging method. The effect has originally been explained by diffraction, and high spatial coherence has been claimed to be a necessary precondition. However, edge enhancement has also been found in conventional imaging with high resolution. In such cases the effects can produce artefacts and hinder quantification. In this letter the edge effects at cylindrical shaped samples and long straight edges have been studied in detail. The enhancement can be explained by refraction and total reflection. Using high-resolution imaging, where spatial resolutions better than 50 μm could be achieved, refraction and total reflection peaks - similar to diffraction patterns - could be separated and distinguished.

  5. SRS station 16.3: high-resolution applications

    CERN Document Server

    Murphy, B M; Golshan, M; Moore, M; Reid, J; Kowalski, G

    2001-01-01

    Station 16.3 is a high-resolution X-ray diffraction beamline at Daresbury Laboratory Synchrotron Radiation Source. The data presented demonstrate the high-resolution available on the station utilising the recently commissioned four-reflection Si 1 1 1 monochromator and three-reflection Si 1 1 1 analyser. For comparison, a reciprocal space map of the two-bounce Si 1 1 1 monochromator and two-bounce analyser is also shown. Operation of the station is illustrated with examples for silicon, and for diamond. Lattice parameter variations were measured with accuracies in the part per million range and lattice tilts at the arc second level (DuMond, Phys. Rev. 52 (1937) 872).

  6. High-resolution nuclear magnetic resonance studies of proteins.

    Science.gov (United States)

    Jonas, Jiri

    2002-03-25

    The combination of advanced high-resolution nuclear magnetic resonance (NMR) techniques with high-pressure capability represents a powerful experimental tool in studies of protein folding. This review is organized as follows: after a general introduction of high-pressure, high-resolution NMR spectroscopy of proteins, the experimental part deals with instrumentation. The main section of the review is devoted to NMR studies of reversible pressure unfolding of proteins with special emphasis on pressure-assisted cold denaturation and the detection of folding intermediates. Recent studies investigating local perturbations in proteins and the experiments following the effects of point mutations on pressure stability of proteins are also discussed. Ribonuclease A, lysozyme, ubiquitin, apomyoglobin, alpha-lactalbumin and troponin C were the model proteins investigated.

  7. Turbine component casting core with high resolution region

    Science.gov (United States)

    Kamel, Ahmed; Merrill, Gary B.

    2014-08-26

    A hollow turbine engine component with complex internal features can include a first region and a second, high resolution region. The first region can be defined by a first ceramic core piece formed by any conventional process, such as by injection molding or transfer molding. The second region can be defined by a second ceramic core piece formed separately by a method effective to produce high resolution features, such as tomo lithographic molding. The first core piece and the second core piece can be joined by interlocking engagement that once subjected to an intermediate thermal heat treatment process thermally deform to form a three dimensional interlocking joint between the first and second core pieces by allowing thermal creep to irreversibly interlock the first and second core pieces together such that the joint becomes physically locked together providing joint stability through thermal processing.

  8. Precision crystal alignment for high-resolution electron microscope imaging

    International Nuclear Information System (INIS)

    Wood, G.J.; Beeching, M.J.

    1990-01-01

    One of the more difficult tasks involved in obtaining quality high-resolution electron micrographs is the precise alignment of a specimen into the required zone. The current accepted procedure, which involves changing to diffraction mode and searching for symmetric point diffraction pattern, is insensitive to small amounts of misalignment and at best qualitative. On-line analysis of the fourier space representation of the image, both for determining and correcting crystal tilt, is investigated. 8 refs., 42 figs

  9. A high resolution large dynamic range TDC circuit implementation

    International Nuclear Information System (INIS)

    Lei Wuhu; Liu Songqiu; Ye Weiguo; Han Hui; Li Pengyu

    2003-01-01

    Time measurement technology is usually used in nuclear experimentation. There are many methods of time measurement. The implementation method of Time to Digital Conversion (TDC) by means of electronic is a classical technology. The range and resolution of TDC is different according with different usage. A wide range and high resolution TDC circuit, including its theory and implementation way, is introduced in this paper. The test result is also given. (authors)

  10. A high resolution large dynamic range TDC circuit implementation

    International Nuclear Information System (INIS)

    Lei Wuhu; Liu Songqiu; Li Pengyu; Han Hui; Ye Yanlin

    2005-01-01

    Time measurement technology is usually used in nuclear experimentation. There are many methods of time measurement. The implementation method of Time to Digital Conversion (TDC) by means of electronics is a classical technology. The range and resolution of TDC is different according with different usage. A wide range and high resolution TDC circuit, including its theory and implementation way, is introduced in this paper. The test result is also given. (authors)

  11. Constraining Stochastic Parametrisation Schemes Using High-Resolution Model Simulations

    Science.gov (United States)

    Christensen, H. M.; Dawson, A.; Palmer, T.

    2017-12-01

    Stochastic parametrisations are used in weather and climate models as a physically motivated way to represent model error due to unresolved processes. Designing new stochastic schemes has been the target of much innovative research over the last decade. While a focus has been on developing physically motivated approaches, many successful stochastic parametrisation schemes are very simple, such as the European Centre for Medium-Range Weather Forecasts (ECMWF) multiplicative scheme `Stochastically Perturbed Parametrisation Tendencies' (SPPT). The SPPT scheme improves the skill of probabilistic weather and seasonal forecasts, and so is widely used. However, little work has focused on assessing the physical basis of the SPPT scheme. We address this matter by using high-resolution model simulations to explicitly measure the `error' in the parametrised tendency that SPPT seeks to represent. The high resolution simulations are first coarse-grained to the desired forecast model resolution before they are used to produce initial conditions and forcing data needed to drive the ECMWF Single Column Model (SCM). By comparing SCM forecast tendencies with the evolution of the high resolution model, we can measure the `error' in the forecast tendencies. In this way, we provide justification for the multiplicative nature of SPPT, and for the temporal and spatial scales of the stochastic perturbations. However, we also identify issues with the SPPT scheme. It is therefore hoped these measurements will improve both holistic and process based approaches to stochastic parametrisation. Figure caption: Instantaneous snapshot of the optimal SPPT stochastic perturbation, derived by comparing high-resolution simulations with a low resolution forecast model.

  12. High-Resolution Imaging of Colliding and Merging Galaxies

    Science.gov (United States)

    Whitmore, Brad

    1991-07-01

    We propose to obtain high-resolution images, using the WF/PC, of two colliding and merging galaxies (i.e., NGC 4038/4039 = "The Antennae" and NGC 7252 ="Atoms-for-Peace Galaxy". Our goal is to use HST to make critical observations of each object in order to gain a better understanding of the various phases of the merger process. Our primary objective is to determine whether globular clusters are formed during mergers\\?

  13. PROFIL-360 high resolution steam generator tube profilometry system

    International Nuclear Information System (INIS)

    Glass, S.W.

    1985-01-01

    A high-resolution profilometry system, PROFIL 360, has been developed to assess the condition of steam generator tubes and rapidly produce the data to evaluate the potential for developing in-service leaks. The probe has an electromechanical sensor in a rotating head. This technique has been demonstrated in the field, saving tubes that would have been plugged with the go-gauge criterion and indicating plugging other high-risk candidates that might otherwise not have been removed from service

  14. Profil-360 high resolution steam generator tube profilometry system

    International Nuclear Information System (INIS)

    Glass, S.W.

    1985-01-01

    A high-resolution profilometry system, PROFIL 360, has been developed to assess the condition of steam generator tubes and rapidly produce the data to evaluate the potential for developing in-service leaks. The probe has an electromechanical sensor in a rotating head. This technique has been demonstrated in the field, saving tubes that would have been plugged with the go-gauge criterion and indicating plugging other high-risk candidates that might otherwise not have been removed from service

  15. Tuberculous otitis media: findings on high-resolution CT

    International Nuclear Information System (INIS)

    Lungenschmid, D.; Buchberger, W.; Schoen, G.; Schoepf, R.; Mihatsch, T.; Birbamer, G.; Wicke, K.

    1993-01-01

    We describe two cases of tuberculous otitis media studied with high-resolution computed tomography (CT). Findings included extensive soft tissue densities with fluid levels in the tympanic cavity, the antrum, the mastoid and petrous air cells. Multifocal bony erosions and reactive bone sclerosis were seen as well. CT proved valuable for planning therapy by accurately displaying the involvement of the various structures of the middle and inner ear. However, the specific nature of the disease could only be presumed. (orig.)

  16. Environmental high resolution electron microscopy and applications to chemical science

    OpenAIRE

    Boyes, Edward; Gai, Pratibha

    2017-01-01

    An environmental cell high resolution electron microscope (EHREM) has been developed for in situ studies of dynamic chemical reactions on the atomic scale. It allows access to metastable intermediate phases of catalysts and to sequences of reversible microstructural and chemical development associated with the activation, deactivation and poisoning of a catalyst. Materials transported through air can be restored or recreated and samples damaged, e.g. by dehydration, by the usual vacuum enviro...

  17. Creating the High-Resolution Settlement Layer - lessons learned

    Science.gov (United States)

    Gros, A.

    2017-12-01

    Facebook publishes the High-resolution Settlement Layer (HRSL: https://ciesin.columbia.edu/data/hrsl/) in collaboration with Columbia University's CIESIN institute and the World Bank. So far, data for 13 countries have been published over the past nine months. HRSL data for Burkina Faso, Ghana, Haiti, Ivory Coast, Madagascar, Malawi, Mexico, The Philippines, Rwanda, South Africa, Sri Lanka, Thailand, and Uganda are available for download. We will present a status update and report on lessons learned.

  18. High-resolution spectroscopy of gases for industrial applications

    OpenAIRE

    Fateev, Alexander; Clausen, Sønnik

    2012-01-01

    High-resolution spectroscopy of gases is a powerful technique which has various fundamental and practical applications: in situ simultaneous measurements of gas temperature and gas composition, radiative transfer modeling, validation of existing and developing of new databases and etc. Existing databases (e.g. HITRAN, HITEMP or CDSD) can normally be used for absorption spectra calculations at limited temperature/pressure ranges. Therefore experimental measurements of absorption/transmission s...

  19. High resolution atomic spectra of rare earths : progress report

    International Nuclear Information System (INIS)

    Saksena, G.D.; Ahmad, S.A.

    1976-01-01

    High resolution studies of atomic spectra of neodymium and gadolinium are being carried out on a recording Fabry-Perot spectrometer. The present progress report concerns work done on new assignments as well as confirmation of recently assigned electronic configurations and evaluation of isotope shifts of energy levels which have been possible from the isotope shift data obtained for several transitions of NdI, NdII and GdI, GdII respectively. (author)

  20. High-resolution CT of lesions of the optic nerve

    International Nuclear Information System (INIS)

    Peyster, R.G.; Hoover, E.D.; Hershey, B.L.; Haskin, M.E.

    1983-01-01

    The optic nerves are well demonstrated by high-resolution computed tomography. Involvement of the optic nerve by optic gliomas and optic nerve sheath meningiomas is well known. However, nonneoplastic processes such as increased intracranial pressure, optic neuritis, Grave ophthalmopathy, and orbital pseudotumor may also alter the appearance of the optic nerve/sheath on computed tomography. Certain clinical and computed tomographic features permit distinction of these nonneoplastic tumefactions from tumors

  1. Detectors for high resolution dynamic positron emission tomography

    International Nuclear Information System (INIS)

    Derenzo, S.E.; Budinger, T.F.; Huesman, R.H.

    1985-01-01

    Tomography is the technique of producing a photographic image of an opaque specimen by transmitting a beam of x-rays or gamma rays through the specimen onto an adjacent photographic film. The image results from variations in thickness, density, and chemical composition, of the specimen. This technique is used to study the metabolism of the human brain. This article examines the design of equipment used for high resolution dynamic positron emission tomography. 27 references, 5 figures, 3 tables

  2. High-Resolution Esophageal Manometry: A Time Motion Study

    Directory of Open Access Journals (Sweden)

    Daniel C Sadowski

    2008-01-01

    Full Text Available INTRODUCTION: High-resolution manometry (HRM of the esophagus is a new technique that provides a more precise assessment of esophageal motility than conventional techniques. Because HRM measures pressure events along the entire length of the esophagus simultaneously, clinical procedure time should be shorter because less catheter manipulation is required. According to manufacturer advertising, the new HRM system is more accurate and up to 50% faster than conventional methods.

  3. High Resolution Mass Spectrometry of Polyfluorinated Polyether-Based Formulation

    DEFF Research Database (Denmark)

    Dimzon, Ian Ken; Trier, Xenia; Frömel, Tobias

    2016-01-01

    High resolution mass spectrometry (HRMS) was successfully applied to elucidate the structure of a polyfluorinated polyether (PFPE)-based formulation. The mass spectrum generated from direct injection into the MS was examined by identifying the different repeating units manually and with the aid o......-fluorinated polymers. The information from MS is essential in studying the physico-chemical properties of PFPEs and can help in assessing the risks they pose to the environment and to human health. Graphical Abstract ᅟ....

  4. Chronic pneumonitis of infancy: high-resolution CT findings

    International Nuclear Information System (INIS)

    Olsen, Oeystein E.; Owens, Catherine M.; Sebire, Neil J.; Jaffe, Adam

    2004-01-01

    Chronic pneumonitis of infancy (CPI) is a very rare entity. We report the chest radiography and high-resolution CT (HRCT) findings in an infant with histopathologically confirmed CPI. The child was admitted for intensive care 18 h after birth and died at 39 days of age. On HRCT there was diffuse ground-glass change, interlobular septal thickening and discrete centrilobular nodules. An accurate diagnosis is crucial for correct management; however, several entities with the same HRCT findings are recognized. (orig.)

  5. Acute pulmonary injury: high-resolution CT and histopathological spectrum

    Science.gov (United States)

    Obadina, E T; Torrealba, J M

    2013-01-01

    Acute lung injury usually causes hypoxaemic respiratory failure and acute respiratory distress syndrome (ARDS). Although diffuse alveolar damage is the hallmark of ARDS, other histopathological patterns of injury, such as acute and fibrinoid organising pneumonia, can be associated with acute respiratory failure. Acute eosinophilic pneumonia can also cause acute hypoxaemic respiratory failure and mimic ARDS. This pictorial essay reviews the high-resolution CT findings of acute lung injury and the correlative histopathological findings. PMID:23659926

  6. Aspects of pulmonary histiocytosis X on high resolution computed tomography

    International Nuclear Information System (INIS)

    Costa, N.S.S.; Castro Lessa Angela, M.T. de; Angelo Junior, J.R.L.; Silva, F.M.D.; Kavakama, J.; Carvalho, C.R.R. de; Cerri, G.G.

    1995-01-01

    Pulmonary histiocytosis X is a disease that occurs in young adults and presents with nodules and cysts, mainly in upper lobes, with consequent pulmonary fibrosis. These pulmonary changes are virtually pathognomonic findings on high resolution computed tomography, that allows estimate the area of the lung involved and distinguish histiocytosis X from other disorders that also produces nodules and cysts. (author). 10 refs, 2 tabs, 6 figs

  7. Concept for a new high resolution high intensity diffractometer

    Energy Technology Data Exchange (ETDEWEB)

    Stuhr, U [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-09-01

    A concept of a new time-of-flight powder-diffractometer for a thermal neutral beam tube at SINQ is presented. The design of the instrument optimises the contradictory conditions of high intensity and high resolution. The high intensity is achieved by using many neutron pulses simultaneously. By analysing the time-angle-pattern of the detected neutrons an assignment of the neutrons to a single pulse is possible. (author) 3 figs., tab., refs.

  8. COST EFFECTIVE AND HIGH RESOLUTION SUBSURFACE CHARACTERIZATION USING HYDRAULIC TOMOGRAPHY

    Science.gov (United States)

    2017-08-01

    objective of this project is to provide the DoD and its remediation contractors with the HT technology for delineating the spatial distribution of...STATEMENT Approved for public release; distribution is unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT Hydraulic Tomography ( HT ) is a high-resolution...performance of subsurface remedial actions at environmental sites. The good technical performance and cost-effectiveness of HT have been demonstrated in

  9. High resolution solar soft X-ray spectrometer

    International Nuclear Information System (INIS)

    Zhang Fei; Wang Huanyu; Peng Wenxi; Liang Xiaohua; Zhang Chunlei; Cao Xuelei; Jiang Weichun; Zhang Jiayu; Cui Xingzhu

    2012-01-01

    A high resolution solar soft X-ray spectrometer (SOX) payload onboard a satellite is developed. A silicon drift detector (SDD) is adopted as the detector of the SOX spectrometer. The spectrometer consists of the detectors and their readout electronics, a data acquisition unit and a payload data handling unit. A ground test system is also developed to test SOX. The test results show that the design goals of the spectrometer system have been achieved. (authors)

  10. High-Resolution Wind Measurements for Offshore Wind Energy Development

    Science.gov (United States)

    Nghiem, Son V.; Neumann, Gregory

    2011-01-01

    A mathematical transform, called the Rosette Transform, together with a new method, called the Dense Sampling Method, have been developed. The Rosette Transform is invented to apply to both the mean part and the fluctuating part of a targeted radar signature using the Dense Sampling Method to construct the data in a high-resolution grid at 1-km posting for wind measurements over water surfaces such as oceans or lakes.

  11. Pneumonia: high-resolution CT findings in 114 patients

    Energy Technology Data Exchange (ETDEWEB)

    Reittner, Pia [Department of Radiology, Vancouver Hospital and Health Sciences Center, 855 W. 12th Ave., Vancouver, BC (Canada); Department of Radiology, Karl Franzens University and University Hospital Graz, Auenbruggerplatz 9, 8036 Graz (Austria); Ward, Suzanne; Heyneman, Laura; Mueller, Nestor L. [Department of Radiology, Vancouver Hospital and Health Sciences Center, 855 W. 12th Ave., Vancouver, BC (Canada); Johkoh, Takeshi [Department of Radiology, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565-0825 (Japan)

    2003-03-01

    The objective of the present study was to assess the high-resolution CT appearances of different types of pneumonia. The high-resolution CT scans obtained in 114 patients (58 immunocompetent, 59 immunocompromised) with bacterial, Mycoplasma pneumoniae, viral, fungal, and Pneumocystis carinii pneumonias were analyzed retrospectively by two independent observers for presence, pattern, and distribution of abnormalities. Areas of air-space consolidation were not detected in patients with viral pneumonia and were less frequently seen in patients with Pneumocystis carinii pneumonia (2 of 22 patients, 9%) than in bacterial (30 of 35, 85%), Mycoplasma pneumoniae (22 of 28, 79%), and fungal pneumonias (15 of 20, 75%; p<0.01). There was no significant difference in the prevalence or distribution of consolidation between bacterial, Mycoplasma pneumoniae, and fungal pneumonias. Extensive symmetric bilateral areas of ground-glass attenuation were present in 21 of 22 (95%) patients with Pneumocystis carinii pneumonia and were not seen in other pneumonias except in association with areas of consolidation and nodules. Centrilobular nodules were present less commonly in bacterial pneumonia (6 of 35 patients, 17%) than in Mycoplasma pneumoniae (24 of 28, 96%), viral (7 of 9, 78%), or fungal (12 of 20, 92%) pneumonia (p<0.01). Except for Pneumocystis carinii pneumonia and Mycoplasma pneumoniae pneumonia, which often have a characteristic appearance, high-resolution CT is of limited value in the differential diagnosis of the various types of infective pneumonia. (orig.)

  12. High-resolution CT findings in Streptococcus milleri pulmonary infection

    International Nuclear Information System (INIS)

    Okada, F.; Ono, A.; Ando, Y.; Nakayama, T.; Ishii, H.; Hiramatsu, K.; Sato, H.; Kira, A.; Otabe, M.; Mori, H.

    2013-01-01

    Aim: To assess pulmonary high-resolution computed tomography (CT) findings in patients with acute Streptococcus milleri pulmonary infection. Materials and methods: Sixty consecutive patients with acute S. milleri pneumonia who had undergone high-resolution CT chest examinations between January 2004 and March 2010 were retrospectively identified. Twenty-seven patients with concurrent infections were excluded. The final study group comprised 33 patients (25 men, 8 women; aged 20–88 years, mean 63.1 years) with S. milleri infection. The patients' clinical findings were assessed. Parenchymal abnormalities, enlarged lymph nodes, and pleural effusion were evaluated on high-resolution CT. Results: Underlying conditions included malignancy (n = 15), a smoking habit (n = 11), and diabetes mellitus (n = 8). CT images of all patients showed abnormal findings, including ground-glass opacity (n = 24), bronchial wall thickening (n = 23), consolidation (n = 17), and cavities (n = 7). Pleural effusion was found in 18 patients, and complex pleural effusions were found in seven patients. Conclusion: Pulmonary infection caused by S. milleri was observed mostly in male patients with underlying conditions such as malignancy or a smoking habit. The CT findings in patients with S. milleri consisted mainly of ground-glass opacity, bronchial wall thickening, pleural effusions, and cavities

  13. Adaptive optics with pupil tracking for high resolution retinal imaging.

    Science.gov (United States)

    Sahin, Betul; Lamory, Barbara; Levecq, Xavier; Harms, Fabrice; Dainty, Chris

    2012-02-01

    Adaptive optics, when integrated into retinal imaging systems, compensates for rapidly changing ocular aberrations in real time and results in improved high resolution images that reveal the photoreceptor mosaic. Imaging the retina at high resolution has numerous potential medical applications, and yet for the development of commercial products that can be used in the clinic, the complexity and high cost of the present research systems have to be addressed. We present a new method to control the deformable mirror in real time based on pupil tracking measurements which uses the default camera for the alignment of the eye in the retinal imaging system and requires no extra cost or hardware. We also present the first experiments done with a compact adaptive optics flood illumination fundus camera where it was possible to compensate for the higher order aberrations of a moving model eye and in vivo in real time based on pupil tracking measurements, without the real time contribution of a wavefront sensor. As an outcome of this research, we showed that pupil tracking can be effectively used as a low cost and practical adaptive optics tool for high resolution retinal imaging because eye movements constitute an important part of the ocular wavefront dynamics.

  14. The high resolution shear wave seismic reflection technique

    International Nuclear Information System (INIS)

    Johnson, W.J.; Clark, J.C.

    1991-04-01

    This report presents the state-of-the-art of the high resolution S-wave reflection technique. Published and unpublished literature has been reviewed and discussions have been held with experts. Result is to confirm that the proposed theoretical and practical basis for identifying aquifer systems using both P- and S-wave reflections is sound. Knowledge of S-wave velocity and P-wave velocity is a powerful tool for assessing the fluid characteristics of subsurface layers. Material properties and lateral changes in material properties such as change from clay to sand, can be inferred from careful dual evaluation of P and S-wave records. The high resolution S-wave reflection technique has seen its greatest application to date as part of geotechnical studies for building foundations in the Far East. Information from this type of study has been evaluated and will be incorporated in field studies. In particular, useful information regarding S-wave sources, noise suppression and recording procedures will be incorporated within the field studies. Case histories indicate that the best type of site for demonstrating the power of the high resolution S-wave technique will be in unconsolidated soil without excessive structural complexities. More complex sites can form the basis for subsequent research after the basic principles of the technique can be established under relatively uncomplicated conditions

  15. Pneumonia: high-resolution CT findings in 114 patients

    International Nuclear Information System (INIS)

    Reittner, Pia; Ward, Suzanne; Heyneman, Laura; Mueller, Nestor L.; Johkoh, Takeshi

    2003-01-01

    The objective of the present study was to assess the high-resolution CT appearances of different types of pneumonia. The high-resolution CT scans obtained in 114 patients (58 immunocompetent, 59 immunocompromised) with bacterial, Mycoplasma pneumoniae, viral, fungal, and Pneumocystis carinii pneumonias were analyzed retrospectively by two independent observers for presence, pattern, and distribution of abnormalities. Areas of air-space consolidation were not detected in patients with viral pneumonia and were less frequently seen in patients with Pneumocystis carinii pneumonia (2 of 22 patients, 9%) than in bacterial (30 of 35, 85%), Mycoplasma pneumoniae (22 of 28, 79%), and fungal pneumonias (15 of 20, 75%; p<0.01). There was no significant difference in the prevalence or distribution of consolidation between bacterial, Mycoplasma pneumoniae, and fungal pneumonias. Extensive symmetric bilateral areas of ground-glass attenuation were present in 21 of 22 (95%) patients with Pneumocystis carinii pneumonia and were not seen in other pneumonias except in association with areas of consolidation and nodules. Centrilobular nodules were present less commonly in bacterial pneumonia (6 of 35 patients, 17%) than in Mycoplasma pneumoniae (24 of 28, 96%), viral (7 of 9, 78%), or fungal (12 of 20, 92%) pneumonia (p<0.01). Except for Pneumocystis carinii pneumonia and Mycoplasma pneumoniae pneumonia, which often have a characteristic appearance, high-resolution CT is of limited value in the differential diagnosis of the various types of infective pneumonia. (orig.)

  16. The demonstration of the auditory ossicles by high resolution CT

    International Nuclear Information System (INIS)

    Lloyd, G.A.S.; Boulay, G.H. du; Phelps, P.D.; Pullicino, P.

    1979-01-01

    The high resolution CT scanning system introduced by EMI in 1978 has added a new dimension to computerised tomography in otology. The apparatus used for this study was an EMI CT 5005 body scanner adapted for head and neck scanning and incorporating a high resolution facility. The latter has proved most advantageous in areas of relatively high differential absorption, so that its application to the demonstration of abnormalities in the petrous temporal bone, and in particular middle ear disease, has been very rewarding. Traumatic ossicular disruptions may now be demonstrated and the high contrast of CT often shows them better than conventional hypocycloidal tomography. The stapes is also better visualised and congenital abnormalities of its superstructure have been recorded. These studies have been achieved with a very acceptable level of radiation to the eye, lens and cornea and the technique is clearly a rival to conventional pluridirectional tomography in the assessment of the petrous temporal bone. With further design improvements high resolution CT could completely replace existing techniques. (orig.) [de

  17. Progress in high-resolution x-ray holographic microscopy

    International Nuclear Information System (INIS)

    Jacobsen, C.; Kirz, J.; Howells, M.; McQuaid, K.; Rothman, S.; Feder, R.; Sayre, D.

    1987-07-01

    Among the various types of x-ray microscopes that have been demonstrated, the holographic microscope has had the largest gap between promise and performance. The difficulties of fabricating x-ray optical elements have led some to view holography as the most attractive method for obtaining the ultimate in high resolution x-ray micrographs; however, we know of no investigations prior to 1987 that clearly demonstrated submicron resolution in reconstructed images. Previous efforts suffered from problems such as limited resolution and dynamic range in the recording media, low coherent x-ray flux, and aberrations and diffraction limits in visible light reconstruction. We have addressed the recording limitations through the use of an undulator x-ray source and high-resolution photoresist recording media. For improved results in the readout and reconstruction steps, we have employed metal shadowing and transmission electron microscopy, along with numerical reconstruction techniques. We believe that this approach will allow holography to emerge as a practical method of high-resolution x-ray microscopy. 30 refs., 4 figs

  18. Toward high-resolution NMR spectroscopy of microscopic liquid samples

    Energy Technology Data Exchange (ETDEWEB)

    Butler, Mark C.; Mehta, Hardeep S.; Chen, Ying; Reardon, Patrick N.; Renslow, Ryan S.; Khbeis, Michael; Irish, Duane; Mueller, Karl T.

    2017-01-01

    A longstanding limitation of high-resolution NMR spectroscopy is the requirement for samples to have macroscopic dimensions. Commercial probes, for example, are designed for volumes of at least 5 mL, in spite of decades of work directed toward the goal of miniaturization. Progress in miniaturizing inductive detectors has been limited by a perceived need to meet two technical requirements: (1) minimal separation between the sample and the detector, which is essential for sensitivity, and (2) near-perfect magnetic-field homogeneity at the sample, which is typically needed for spectral resolution. The first of these requirements is real, but the second can be relaxed, as we demonstrate here. By using pulse sequences that yield high-resolution spectra in an inhomogeneous field, we eliminate the need for near-perfect field homogeneity and the accompanying requirement for susceptibility matching of microfabricated detector components. With this requirement removed, typical imperfections in microfabricated components can be tolerated, and detector dimensions can be matched to those of the sample, even for samples of volume << 5 uL. Pulse sequences that are robust to field inhomogeneity thus enable small-volume detection with optimal sensitivity. We illustrate the potential of this approach to miniaturization by presenting spectra acquired with a flat-wire detector that can easily be scaled to subnanoliter volumes. In particular, we report high-resolution NMR spectroscopy of an alanine sample of volume 500 pL.

  19. Progress in high-resolution x-ray holographic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jacobsen, C.; Kirz, J.; Howells, M.; McQuaid, K.; Rothman, S.; Feder, R.; Sayre, D.

    1987-07-01

    Among the various types of x-ray microscopes that have been demonstrated, the holographic microscope has had the largest gap between promise and performance. The difficulties of fabricating x-ray optical elements have led some to view holography as the most attractive method for obtaining the ultimate in high resolution x-ray micrographs; however, we know of no investigations prior to 1987 that clearly demonstrated submicron resolution in reconstructed images. Previous efforts suffered from problems such as limited resolution and dynamic range in the recording media, low coherent x-ray flux, and aberrations and diffraction limits in visible light reconstruction. We have addressed the recording limitations through the use of an undulator x-ray source and high-resolution photoresist recording media. For improved results in the readout and reconstruction steps, we have employed metal shadowing and transmission electron microscopy, along with numerical reconstruction techniques. We believe that this approach will allow holography to emerge as a practical method of high-resolution x-ray microscopy. 30 refs., 4 figs.

  20. Global Monitoring of Mountain Glaciers Using High-Resolution Spotlight Imaging from the International Space Station

    Science.gov (United States)

    Donnellan, A.; Green, J. J.; Bills, B. G.; Goguen, J.; Ansar, A.; Knight, R. L.; Hallet, B.; Scambos, T. A.; Thompson, L. G.; Morin, P. J.

    2013-12-01

    Mountain glaciers around the world are retreating rapidly, contributing about 20% to present-day sea level rise. Numerous studies have shown that mountain glaciers are sensitive to global environmental change. Temperate-latitude glaciers and snowpack provide water for over 1 billion people. Glaciers are a resource for irrigation and hydroelectric power, but also pose flood and avalanche hazards. Accurate mass balance assessments have been made for only 280 glaciers, yet there are over 130,000 in the World Glacier Inventory. The rate of glacier retreat or advance can be highly variable, is poorly sampled, and inadequately understood. Liquid water from ice front lakes, rain, melt, or sea water and debris from rocks, dust, or pollution interact with glacier ice often leading to an amplification of warming and further melting. Many mountain glaciers undergo rapid and episodic events that greatly change their mass balance or extent but are sparsely documented. Events include calving, outburst floods, opening of crevasses, or iceberg motion. Spaceborne high-resolution spotlight optical imaging provides a means of clarifying the relationship between the health of mountain glaciers and global environmental change. Digital elevation models (DEMs) can be constructed from a series of images from a range of perspectives collected by staring at a target during a satellite overpass. It is possible to collect imagery for 1800 targets per month in the ×56° latitude range, construct high-resolution DEMs, and monitor changes in high detail over time with a high-resolution optical telescope mounted on the International Space Station (ISS). Snow and ice type, age, and maturity can be inferred from different color bands as well as distribution of liquid water. Texture, roughness, albedo, and debris distribution can be estimated by measuring bidirectional reflectance distribution functions (BRDF) and reflectance intensity as a function of viewing angle. The non-sun-synchronous orbit

  1. Direct fabrication of high-resolution three-dimensional polymeric scaffolds using electrohydrodynamic hot jet plotting

    International Nuclear Information System (INIS)

    Wei, Chuang; Dong, Jingyan

    2013-01-01

    This paper presents the direct three-dimensional (3D) fabrication of polymer scaffolds with sub-10 µm structures using electrohydrodynamic jet (EHD-jet) plotting of melted thermoplastic polymers. Traditional extrusion-based fabrication approaches of 3D periodic porous structures are very limited in their resolution, due to the excessive pressure requirement for extruding highly viscous thermoplastic polymers. EHD-jet printing has become a high-resolution alternative to other forms of nozzle deposition-based fabrication approaches by generating micro-scale liquid droplets or a fine jet through the application of a large electrical voltage between the nozzle and the substrate. In this study, we successfully apply EHD-jet plotting technology with melted biodegradable polymer (polycaprolactone, or PCL) for the fabrication of 2D patterns and 3D periodic porous scaffold structures in potential tissue engineering applications. Process conditions (e.g. electrical voltage, pressure, plotting speed) have been thoroughly investigated to achieve reliable jet printing of fine filaments. We have demonstrated for the first time that the EHD-jet plotting process is capable of the fabrication of 3D periodic structures with sub-10 µm resolution, which has great potential in advanced biomedical applications, such as cell alignment and guidance. (paper)

  2. High resolution solar observations from first principles to applications

    Science.gov (United States)

    Verdoni, Angelo P.

    2009-10-01

    The expression "high-resolution observations" in Solar Physics refers to the spatial, temporal and spectral domains in their entirety. High-resolution observations of solar fine structure are a necessity to answer many of the intriguing questions related to solar activity. However, a researcher building instruments for high-resolution observations has to cope with the fact that these three domains often have diametrically opposed boundary conditions. Many factors have to be considered in the design of a successful instrument. Modern post-focus instruments are more closely linked with the solar telescopes that they serve than in past. In principle, the quest for high-resolution observations already starts with the selection of the observatory site. The site survey of the Advanced Technology Solar Telescope (ATST) under the stewardship of the National Solar Observatory (NSO) has identified Big Bear Solar Observatory (BBSO) as one of the best sites for solar observations. In a first step, the seeing characteristics at BBSO based on the data collected for the ATST site survey are described. The analysis will aid in the scheduling of high-resolution observations at BBSO as well as provide useful information concerning the design and implementation of a thermal control system for the New Solar Telescope (NST). NST is an off-axis open-structure Gregorian-style telescope with a 1.6 m aperture. NST will be housed in a newly constructed 5/8-sphere ventilated dome. With optics exposed to the surrounding air, NST's open-structure design makes it particularly vulnerable to the effects of enclosure-related seeing. In an effort to mitigate these effects, the initial design of a thermal control system for the NST dome is presented. The goal is to remediate thermal related seeing effects present within the dome interior. The THermal Control System (THCS) is an essential component for the open-telescope design of NST to work. Following these tasks, a calibration routine for the

  3. A subspace approach to high-resolution spectroscopic imaging.

    Science.gov (United States)

    Lam, Fan; Liang, Zhi-Pei

    2014-04-01

    To accelerate spectroscopic imaging using sparse sampling of (k,t)-space and subspace (or low-rank) modeling to enable high-resolution metabolic imaging with good signal-to-noise ratio. The proposed method, called SPectroscopic Imaging by exploiting spatiospectral CorrElation, exploits a unique property known as partial separability of spectroscopic signals. This property indicates that high-dimensional spectroscopic signals reside in a very low-dimensional subspace and enables special data acquisition and image reconstruction strategies to be used to obtain high-resolution spatiospectral distributions with good signal-to-noise ratio. More specifically, a hybrid chemical shift imaging/echo-planar spectroscopic imaging pulse sequence is proposed for sparse sampling of (k,t)-space, and a low-rank model-based algorithm is proposed for subspace estimation and image reconstruction from sparse data with the capability to incorporate prior information and field inhomogeneity correction. The performance of the proposed method has been evaluated using both computer simulations and phantom studies, which produced very encouraging results. For two-dimensional spectroscopic imaging experiments on a metabolite phantom, a factor of 10 acceleration was achieved with a minimal loss in signal-to-noise ratio compared to the long chemical shift imaging experiments and with a significant gain in signal-to-noise ratio compared to the accelerated echo-planar spectroscopic imaging experiments. The proposed method, SPectroscopic Imaging by exploiting spatiospectral CorrElation, is able to significantly accelerate spectroscopic imaging experiments, making high-resolution metabolic imaging possible. Copyright © 2014 Wiley Periodicals, Inc.

  4. Landslide detection using very high-resolution satellite imageries

    Science.gov (United States)

    Suga, Yuzo; Konishi, Tomohisa

    2012-10-01

    The heavy rain induced by the 12th typhoon caused landslide disaster at Kii Peninsula in the middle part of Japan. We propose a quick response method for landslide disaster mapping using very high resolution (VHR) satellite imageries. Especially, Synthetic Aperture Radar (SAR) is effective because it has the capability of all weather and day/night observation. In this study, multi-temporal COSMO-SkyMed imageries were used to detect the landslide areas. It was difficult to detect the landslide areas using only backscatter change pattern derived from pre- and post-disaster COSMOSkyMed imageries. Thus, the authors adopted a correlation analysis which the moving window was selected for the correlation coefficient calculation. Low value of the correlation coefficient reflects land cover change between pre- and post-disaster imageries. This analysis is effective for the detection of landslides using SAR data. The detected landslide areas were compared with the area detected by EROS-B high resolution optical image. In addition, we have developed 3D viewing system for geospatial visualizing of the damaged area using these satellite image data with digital elevation model. The 3D viewing system has the performance of geographic measurement with respect to elevation height, area and volume calculation, and cross section drawing including landscape viewing and image layer construction using a mobile personal computer with interactive operation. As the result, it was verified that a quick response for the detection of landslide disaster at the initial stage could be effectively performed using optical and SAR very high resolution satellite data by means of 3D viewing system.

  5. Proceedings of the workshop on high resolution computed microtomography (CMT)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-02-01

    The purpose of the workshop was to determine the status of the field, to define instrumental and computational requirements, and to establish minimum specifications required by possible users. The most important message sent by implementers was the remainder that CMT is a tool. It solves a wide spectrum of scientific problems and is complementary to other microscopy techniques, with certain important advantages that the other methods do not have. High-resolution CMT can be used non-invasively and non-destructively to study a variety of hierarchical three-dimensional microstructures, which in turn control body function. X-ray computed microtomography can also be used at the frontiers of physics, in the study of granular systems, for example. With high-resolution CMT, for example, three-dimensional pore geometries and topologies of soils and rocks can be obtained readily and implemented directly in transport models. In turn, these geometries can be used to calculate fundamental physical properties, such as permeability and electrical conductivity, from first principles. Clearly, use of the high-resolution CMT technique will contribute tremendously to the advancement of current R and D technologies in the production, transport, storage, and utilization of oil and natural gas. It can also be applied to problems related to environmental pollution, particularly to spilling and seepage of hazardous chemicals into the Earth's subsurface. Applications to energy and environmental problems will be far-ranging and may soon extend to disciplines such as materials science--where the method can be used in the manufacture of porous ceramics, filament-resin composites, and microelectronics components--and to biomedicine, where it could be used to design biocompatible materials such as artificial bones, contact lenses, or medication-releasing implants. Selected papers are indexed separately for inclusion in the Energy Science and Technology Database.

  6. High Resolution Thz and FIR Spectroscopy of SOCl_2

    Science.gov (United States)

    Martin-Drumel, M. A.; Cuisset, A.; Sadovskii, D. A.; Mouret, G.; Hindle, F.; Pirali, O.

    2013-06-01

    Thionyl chloride (SOCl_2) is an extremely powerful oxidant widely used in industrial processes and playing a role in the chemistry of the atmosphere. In addition, it has a molecular configuration similar to that of phosgene (COCl_2), and is therefore of particular interest for security and defense applications. Low resolution vibrational spectra of gas phase SOCl_2 as well as high resolution pure rotational transitions up to 25 GHz have previously been investigated. To date no high resolution data are reported at frequencies higher than 25 GHz. We have investigated the THz absorption spectrum of SOCl_2 in the spectral region 70-650 GHz using a frequency multiplier chain coupled to a 1 m long single path cell containing a pressure of about 15 μbar. At the time of the writing, about 8000 pure rotational transitions of SO^{35}Cl_2 with highest J and K_a values of 110 and 50 respectively have been assigned on the spectrum. We have also recorded the high resolution FIR spectra of SOCl_2 in the spectral range 50-700 wn using synchrotron radiation at the AILES beamline of SOLEIL facility. A White-type cell aligned with an absorption path length of 150 m has been used to record, at a resolution of 0.001 wn, two spectra at pressures of 5 and 56 μbar of SOCl_2. On these spectra all FIR modes of SOCl_2 are observed (ν_2 to ν_6) and present a resolved rotational structure. Their analysis is in progress. T. J. Johnson et al., J. Phys. Chem. A 107, 6183 (2003) D. E. Martz and R. T. Lagemann, J. Chem. Phys. 22,1193 (1954) H. S. P. Müller and M. C. L. Gerry, J. Chem. Soc. Faraday Trans. 90, 3473 (1994)

  7. Geological survey by high resolution electrical survey on granite areas

    International Nuclear Information System (INIS)

    Sugimoto, Yoshihiro; Yamada, Naoyuki

    2002-03-01

    As an Integral part of the geological survey in 'The study of the regions ground water flow system' that we are carrying out with Tono Geoscience Center, we proved the relation between the uncontinuation structure such as lineament in the base rock and resistivity structure (resistivity distribution), for the purpose of that confirms the efficacy of the high resolution electrical survey as geological survey, we carried out high resolution electrical survey on granite area. We obtained the following result, by the comparison of resistivity distribution with established geological survey, lineament analysis and investigative drilling. 1. The resistivity structure of this survey area is almost able to classify it into the following four range. 1) the low resistivity range of 50-800 Ωm, 2) The resistivity range like the middle of 200-2000 Ωm, 3) The high resistivity range of 2000 Ωm over, 4) The low resistivity range of depth of the survey line 400-550 section. 2. The low resistivity range of 4) that correspond with the established geological data is not admitted. 3. It was confirmed that resistivity structure almost correspond to geological structure by the comparison with the established data. 4. The small-scale low resistivity area is admitted in the point equivalent to the lineament position of established. 5. We carried out it with the simulation method about the low resistivity range of 4). As a result, it understood that it has the possibility that the narrow ratio low resistivity area is shown as the wide ratio resistivity range in the analysis section. In the survey in this time, it is conceivable that the resistivity distribution with the possibility of the unhomogeneous and uncontinuation structure of the base rock is being shown conspicuously, the efficacy of the high resolution resistivity survey as geological survey on granite was shown. (author)

  8. High-resolution ultrasonography in assessing temporomandibular joint disc position.

    Science.gov (United States)

    Talmaceanu, Daniel; Lenghel, Lavinia Manuela; Bolog, Nicolae; Popa Stanila, Roxana; Buduru, Smaranda; Leucuta, Daniel Corneliu; Rotar, Horatiu; Baciut, Mihaela; Baciut, Grigore

    2018-02-04

    The purpose of this study was to determine the diagnostic value of high-resolution ultrasonography (US) in temporomandibular joint (TMJ) disc displacements. A number of 74 patients (148 TMJs) with signs and symptoms of TMJ disorders, according to the Research Diagnostic Criteria for Temporomandibular Disorders, were included in this study. All patients received US and magnetic resonance imaging (MRI) of both TMJs 1 to 5 days after the clinical examination. MRI examinations were performed using 1.5 T MRI equipment (Siemens Avanto, Siemens, Erlangen). Ultrasonographic examination was performed on a Hitachi EUB 8500 (Hitachi Medical Corp., Tokyo, Japan) scanner with L 54 M6.5-13 MHz linear transducer. MRI depicted 68 (45.95%) normal joints, 47 (31.76%) with disc displacement with reduction, 33 (22.3%) with disc displacement without reduction and 34 (22.97%) with degenerative changes. US detected 78 (52.7%) normal joints, 37 (25%) with disc displacement with reduction, 33 (22.3%) with disc displacement without reduction and 21 (14.19%) with degenerative changes. Compared to MRI, US showed a sensitivity of 93.1%, specificity of 87.88%, accuracy of 90.32%, a positive predictive value of 87.1% and a negative predictive value of 93.55% for overall diagnosis of disc displacement. The Youden index was 0.81. Based on our results, high-resolution ultrasonography showed high sensitivity, specificity and accuracy in the diagnosis of TMJ disc displacement. It could be a valuable imaging technique in assessing TMJ disc position. The diagnostic value of high-resolution ultrasonography depends strictly on the examiner's skills and on the equipment used.

  9. On temporal correlations in high-resolution frequency counting

    OpenAIRE

    Dunker, Tim; Hauglin, Harald; Rønningen, Ole Petter

    2016-01-01

    We analyze noise properties of time series of frequency data from different counting modes of a Keysight 53230A frequency counter. We use a 10 MHz reference signal from a passive hydrogen maser connected via phase-stable Huber+Suhner Sucoflex 104 cables to the reference and input connectors of the counter. We find that the high resolution gap-free (CONT) frequency counting process imposes long-term correlations in the output data, resulting in a modified Allan deviation that is characteristic...

  10. High Resolution Spectrometer (HRS) particle-identification system

    International Nuclear Information System (INIS)

    Pratt, J.C.; Spencer, J.E.; Whitten, C.A.

    1977-08-01

    The functions of the particle-identification system (PIDS) designed for the High Resolution Spectrometer facility (HRS) at LAMPF are described, together with the mechanical layout, counter hardware, and associated electronics. The system was designed for easy use and to be applicable to currently proposed experiments at HRS. The several strobe signals that can be generated correspond to different event types or characteristics, and logic configuration and timing can be remotely controlled by computer. Concepts of discrete pattern recognition and multidimensional, analog pulse discrimination are used to distinguish between different event types

  11. Computing with high-resolution upwind schemes for hyperbolic equations

    International Nuclear Information System (INIS)

    Chakravarthy, S.R.; Osher, S.; California Univ., Los Angeles)

    1985-01-01

    Computational aspects of modern high-resolution upwind finite-difference schemes for hyperbolic systems of conservation laws are examined. An operational unification is demonstrated for constructing a wide class of flux-difference-split and flux-split schemes based on the design principles underlying total variation diminishing (TVD) schemes. Consideration is also given to TVD scheme design by preprocessing, the extension of preprocessing and postprocessing approaches to general control volumes, the removal of expansion shocks and glitches, relaxation methods for implicit TVD schemes, and a new family of high-accuracy TVD schemes. 21 references

  12. Achieving High Resolution Timer Events in Virtualized Environment.

    Science.gov (United States)

    Adamczyk, Blazej; Chydzinski, Andrzej

    2015-01-01

    Virtual Machine Monitors (VMM) have become popular in different application areas. Some applications may require to generate the timer events with high resolution and precision. This however may be challenging due to the complexity of VMMs. In this paper we focus on the timer functionality provided by five different VMMs-Xen, KVM, Qemu, VirtualBox and VMWare. Firstly, we evaluate resolutions and precisions of their timer events. Apparently, provided resolutions and precisions are far too low for some applications (e.g. networking applications with the quality of service). Then, using Xen virtualization we demonstrate the improved timer design that greatly enhances both the resolution and precision of achieved timer events.

  13. Free radicals. High-resolution spectroscopy and molecular structure

    International Nuclear Information System (INIS)

    Hirota, E.

    1983-01-01

    High-resolution, high-sensitivity spectroscopy using CW laser and microwave sources has been applied to free radicals and transient molecules to establish their existence and to explore their properties in detail. The radicals studied were mainly generated by discharge-induced reactions. A few molecules are used as typical examples to illustrate the results so far obtained. The molecular and electronic structures of free radicals, intramolecular motions of large amplitudes in some labile molecules, and metastable electronic states of carbenes are given special emphasis. The significance of the present spectroscopic results in other related fields such as astronomy and atmospheric chemistry is stressed. 4 figures, 3 tables

  14. High resolution spectroscopy of six new extreme helium stars

    Science.gov (United States)

    Heber, U.; Jones, G.; Drilling, J. S.

    1986-01-01

    High resolution spectra of six newly discovered extreme helium stars are presented. LSS 5121 is shown to be a spectroscopical twin of the hot extreme helium star HD 160641. A preliminary LTE analysis of LSS 3184 yielded an effective temperature of 22,000 K and a surface gravity of log g = 3.2. Four stars form a new subgroup, classified by sharp-lined He I spectra and pronounced O II spectra, and it is conjectured that these lie close to the Eddington limit. The whole group of extreme helium stars apparently is inhomogeneous with respect to luminosity to mass ratio and chemical composition.

  15. High resolution laser spectroscopy as a diagnostic tool in beams

    International Nuclear Information System (INIS)

    Bergmann, K.; Hefter, U.; Hering, P.

    1977-01-01

    The combination of high resolution laser spectroscopy with the technique of molecular beams allows a very detailed beam research since molecules or atoms in specific quantum states can be sampled yielding previously unavailable sources of data. In these experiments a Na/Na 2 beam emerges from a 0.2 mm nozzle and is collimated by a 2 mm wide slit 50 cm downstream. To probe the molecules a single mode Ar + -laser was used which can be tuned within the gain profile of the laser line (8 GHz) to several transitions between specific levels in the ground state and second electronically excited state of the Na 2 molecule. (Auth.)

  16. High resolution skin colorimetry, strain mapping and mechanobiology.

    Science.gov (United States)

    Devillers, C; Piérard-Franchimont, C; Schreder, A; Docquier, V; Piérard, G E

    2010-08-01

    Skin colours are notoriously different between individuals. They are governed by ethnicities and phototypes, and further influenced by a variety of factors including photoexposures and sustained mechanical stress. Indeed, mechanobiology is a feature affecting the epidermal melanization. High-resolution epiluminescence colorimetry helps in deciphering the effects of forces generated by Langer's lines or relaxed skin tension lines on the melanocyte activity. The same procedure shows a prominent laddering pattern of melanization in striae distensae contrasting with the regular honeycomb pattern in the surrounding skin.

  17. Diesel characterization by high-resolution mass spectrometry - gas chromatography

    International Nuclear Information System (INIS)

    Baldrich, C.A

    1998-01-01

    High-resolution mass spectrometry-gas chromatography is combined with the HC22 method in order to obtain detailed information about the chemical composition of diesel and the distribution of different compound types in terms of its final boiling temperature from a single analysis. The total time elapsed from sample injection and signal processing to obtain final results is 90 minutes. This fact makes this methodology a new and very important tool for the decision making process concerning the most suitable final boiling temperature and the type of treatment of the product in order to obtain diesel that fulfills the international standards. The consistency and repeatability of the experimental results are demonstrated

  18. New detector developments for high resolution positron emission tomography

    International Nuclear Information System (INIS)

    Ziegler, S.I.; Pichler, B.; Lorenz, E.

    1998-01-01

    The strength of quantitative, functional imaging using positron emission tomography, specially in small animals, is limited due to the spatial resolution. Therefore, various tomograph designs employing new scintillators, light sensors, or coincidence electronic are investigated to improve resolution without losses in sensitivity. Luminous scintillators with short light decay time in combination with novel readout schemes using photomultipliers or semiconductor detectors are currently tested by several groups and are implemented in tomographs for small animals. This review summarises the state of development in high resolution positron emission tomography with a detailed description of a system incorporating avalanche photodiode arrays and small scintillation crystals. (orig.) [de

  19. High resolution upgrade of the ATF damping ring BPM system

    International Nuclear Information System (INIS)

    Terunuma, N.; Urakawa, J.; Frisch, J.; May, J.; McCormick, D.; Nelson, J.; Seryi, A.; Smith, T.; Woodley, M.; Briegel, C.; Dysert, R.

    2008-01-01

    A beam position monitor (BPM) upgrade at the KEK Accelerator Test Facility (ATF) damping ring has been accomplished in its first stage, carried out by a KEK/FNAL/SLAC collaboration under the umbrella of the global ILC R and D effort. The upgrade consists of a high resolution, high reproducibility read-out system, based on analog and digital downconversion techniques, digital signal processing, and also tests a new automatic gain error correction schema. The technical concept and realization, as well as preliminary results of beam studies are presented

  20. High-resolution neutron-diffraction measurements to 8 kbar

    Science.gov (United States)

    Bull, C. L.; Fortes, A. D.; Ridley, C. J.; Wood, I. G.; Dobson, D. P.; Funnell, N. P.; Gibbs, A. S.; Goodway, C. M.; Sadykov, R.; Knight, K. S.

    2017-10-01

    We describe the capability to measure high-resolution neutron powder diffraction data to a pressure of at least 8 kbar. We have used the HRPD instrument at the ISIS neutron source and a piston-cylinder design of pressure cell machined from a null-scattering titanium zirconium alloy. Data were collected under hydrostatic conditions from an elpasolite perovskite La?NiMnO?; by virtue of a thinner cell wall on the incident-beam side of the cell, it was possible to obtain data in the instrument's highest resolution back-scattering detector banks up to a maximum pressure of 8.5 kbar.

  1. Diagnosis of cholesteatoma by high resolution computed tomography

    International Nuclear Information System (INIS)

    Kakitsubata, Yousuke; Kakitsubata, Sachiko; Ogata, Noboru; Asada, Keiko; Watanabe, Katsushi; Tohno, Tetsuya; Makino, Kohji

    1988-01-01

    Three normal volunteers and 57 patients with cholesteatoma were examined by high resolution computed tomography. Serial sections were made through the temporal bone at the nasaly inclined position of 30 degree to the orbitomeatal line (semiaxial plane ; SAP). The findings of temporal bone structures in normal subjects were evaluated in SAP and axial plane (OM). Although the both planes showed good visualization, SAP showed both the eustachian tube and tympanic cavity in one slice. In cholesteatoma soft tissue masses in the tympanic cavity, mastoid air cells and Eustachian tube were demonstrated clearly by SAP. (author)

  2. High resolution Moessbauer spectroscopy with 67Zn in metallic systems

    International Nuclear Information System (INIS)

    Potzel, W.

    1985-01-01

    Moessbauer experiments on metallic systems are described where the high resolution 93.3 keV resonance in 67 Zn is used. In the first part, the Cu-Zn alloy system is investigated and the high energy resolution of this Moessbauer transition is employed to determine small changes of the s-electron density at the 67 Zn nucleus when the Zn concentration is changed. In the second part, Zn metal is taken as an example to demonstrate that the 93.3 keV transition is also extremely sensitive to small changes of lattice dynamical effects. 7 refs., 18 figs. (author)

  3. Evaluation of high-resolution CT after tympanoplasty

    International Nuclear Information System (INIS)

    Torizuka, T.; Hayakawa, K.; Sato, Y.; Tanaka, F.; Okuno, Y.

    1991-01-01

    This paper reports on the condition of the middle ear cavity following tympanoplasty which is always of great interest to radiologists and otosurgeons. This study consisted of 21 patients who had various types of tympanoplasty (types I-IV) for chronic otitis media and cholesteatoma by using high-resolution CT (HRCT). HRCT following tympanoplasty was a valuable method for assessing the middle ear aeration and the state of ossicular reconstruction, including stapes prosthesis, although in some cases of soft-tissue mass in the middle ear it was necessary to correlate with clinical findings in order to differentiate between granulation and recurrence

  4. High resolution X-ray spectroscopy of thermal plasmas

    International Nuclear Information System (INIS)

    Canizares, C.R.

    1990-01-01

    This paper concentrates on reviewing highlights of the Focal Plane Crystal Spectrometer (FPCS) results on thermal plasmas, particularly supernova remnants (SNRs) and clusters of galaxies from the Einstein observatory. During Einstein's short but happy life, we made over 400 observations with the FPCS of 40 different objects. Three quarters of these were objects in which the emission was primarily from optically thin thermal plasma, primarily supernova remnants (SNRs) and clusters of galaxies. Thermal plasmas provide an excellent illustration of how spectral data, particularly high resolution spectral data, can be an important tool for probing the physical properties of astrophysical objects. (author)

  5. High-resolution CT of lymphoid interstitial pneumonia

    International Nuclear Information System (INIS)

    Vilgrain, V.; Frija, J.; Yana, C.; Couderc, L.J.; David, M.; Clauvel, J.P.; Laval-Jeantet, M.

    1989-01-01

    Three patients with lymphoid interstitial pneumonia (two HIV 1+ patients with chronic lymphadenopathic syndromes and one with a not-characterized autoimmune disease) have been studied with high-resolution computed tomography (HR-CT). This technique reveals septal lines, small reticulonodular opacities, polyhedral micronodular opacities, 'ground-glass' opacities and a dense, subpleural, curved broken line in one patient. The lesions dominate in the bases of the lungs. They are not characteristic for lymphoid interstitial pneumonia. If a patient presents with a chronic lymphadenopathic syndrome, the diagnosis of an opportunistic infection should not be automatically made, since the syndrome can be caused by lymphoid interstitial pneumonia [fr

  6. Intercalibration of the ZEUS high resolution and backing calorimeters

    International Nuclear Information System (INIS)

    Abramowicz, H.; Czyrkowski, H.; Derlicki, A.; Krzyzanowski, M.; Kudla, I.; Kusmierz, W.; Nowak, R.J.; Pawlak, J.M.; Rajca, A.; Stopczynski, A.; Walczak, R.; Zarnecki, A.F.; Kowalski, T.Z.

    1991-07-01

    We have studied the combined performance of two calorimeters, the high resolution uranium-scintillator prototype of the ZEUS forward calorimeter (FCAL), followed by a prototype of the coarser ZEUS backing calorimeter (BAC), made out of thick iron plates interleaved with planes of aluminium proportional chambers. The test results, obtained in an exposure of the calorimeter system to a hadron test beam at the CERN-SPS, show that the backing calorimeter does fulfil its role of recognizing the energy leaking out of the FCAL calorimeter. The measurement of this energy is feasible, if an appropriate calibration of the BAC calorimeter is performed. (orig.)

  7. High-resolution seismic wave propagation using local time stepping

    KAUST Repository

    Peter, Daniel

    2017-03-13

    High-resolution seismic wave simulations often require local refinements in numerical meshes to accurately capture e.g. steep topography or complex fault geometry. Together with explicit time schemes, this dramatically reduces the global time step size for ground-motion simulations due to numerical stability conditions. To alleviate this problem, local time stepping (LTS) algorithms allow an explicit time stepping scheme to adapt the time step to the element size, allowing nearoptimal time steps everywhere in the mesh. This can potentially lead to significantly faster simulation runtimes.

  8. High resolution resistivity measurements at the Down Ampney research site

    International Nuclear Information System (INIS)

    Hallam, J.R.; Jackson, P.D.; Rainsbury, M.; Raines, M.

    1991-01-01

    A new high resolution resistivity surveying method is described for fault detection and characterisation. The resolution is shown to be significantly higher than conventional apparent resistivity profiling when applied to geological discontinuities such as faults. Nominal fault locations have been determined to an accuracy of 0.5 m, as proven by drilling. Two dimensional profiling and image enhancement of the resulting 2-D data set indicated the possibility of subsidiary fractures and/or lateral changes within the clay to clay' fault zone. The increased resolution allows greater confidence to be placed on both the fault detection and lateral perturbations derived from processed resistance and resistivity images. (Author)

  9. High resolution SPM imaging of organic molecules with functionalized tips

    Czech Academy of Sciences Publication Activity Database

    Jelínek, Pavel

    2017-01-01

    Roč. 29, č. 34 (2017), 1-18, č. článku 343002. ISSN 0953-8984 R&D Projects: GA MŠk LM2015087; GA MŠk 8E15B010; GA ČR(CZ) GC14-16963J Grant - others:AV ČR(CZ) Praemium Academiae Institutional support: RVO:68378271 Keywords : atomic- force microscopy * scanning tunneling microscope * on-surface synthesis * single-molecule * AFM * STM * high resolution * molecules * surfaces Subject RIV: BM - Solid Matter Physics ; Magnetism OBOR OECD: Condensed matter physics (including formerly solid state physics, supercond.) Impact factor: 2.649, year: 2016

  10. High-resolution x-ray photoemission spectra of silver

    DEFF Research Database (Denmark)

    Barrie, A.; Christensen, N. E.

    1976-01-01

    An electron spectrometer fitted with an x-ray monochromator for Al Kα1,2 radiation (1486.6 eV) has been used to record high-resolution x-ray photoelectron spectra for the 4d valence band as well as the 3d spin doublet in silver. The core-level spectrum has a line shape that can be described...... successfully in terms of the many-body theory of Mahan, Nozières, and De Dominicis. The 4d spectrum agrees well with predictions based on a relativistic-augmented-plane-wave band-structure calculation....

  11. Intercalibration of the ZEUS high resolution and backing calorimeters

    International Nuclear Information System (INIS)

    Abramowicz, H.; Czyrkowski, H.; Derlicki, A.; Krzyzanowski, M.; Kudla, I.; Kusmierz, W.; Nowak, R.J.; Pawlak, J.M.; Rajca, A.; Stopczynski, A.; Walczak, R.; Zarnecki, A.F.; Kowalski, T.Z.

    1992-01-01

    We have studied the combined performance of two calorimeters, the high resolution uranium-scintillator prototype of the ZEUS forward calorimeter (FCAL), followed by a prototype of the coarser ZEUS backing calorimeter (BAC), made out of thick iron plates interleaved with planes of aluminium proportional chambers. The test results, obtained in an exposure of the calorimeter system to a hadron test beam at the CERN SPS, show that the backing calorimeter does fulfil its role of recognizing the energy leaking out of the FCAL calorimeter. The measurement of this energy is feasible, if an appropriate calibration of the BAC calorimeter is performed. (orig.)

  12. Clickstream data yields high-resolution maps of science

    Energy Technology Data Exchange (ETDEWEB)

    Bollen, Johan [Los Alamos National Laboratory; Van De Sompel, Herbert [Los Alamos National Laboratory; Hagberg, Aric [Los Alamos National Laboratory; Bettencourt, Luis [Los Alamos National Laboratory; Chute, Ryan [Los Alamos National Laboratory; Rodriguez, Marko A [Los Alamos National Laboratory; Balakireva, Lyudmila [Los Alamos National Laboratory

    2009-01-01

    Intricate maps of science have been created from citation data to visualize the structure of scientific activity. However, most scientific publications are now accessed online. Scholarly web portals record detailed log data at a scale that exceeds the number of all existing citations combined. Such log data is recorded immediately upon publication and keeps track of the sequences of user requests (clickstreams) that are issued by a variety of users across many different domains. Given these advantagees of log datasets over citation data, we investigate whether they can produce high-resolution, more current maps of science.

  13. High resolution X-ray spectroscopy in light antiprotonic atoms

    CERN Document Server

    Borchert, G L; Augsburger, M A; Castelli, C M; Chatellard, D; Egger, J P; El-Khoury, P; Elble, M; Gorke, H; Gotta, D; Hauser, P R; Indelicato, P J; Kirch, K; Lenz, S; Nelms, N; Rashid, K; Schult, O W B; Siems, T; Simons, L M

    2000-01-01

    At the LEAR facility, CERN, antiprotonic L alpha transitions in light elements have been investigated with a focussing crystal spectrometer. The high resolution of the experiment allowed for the first time to resolve in pH/pH the 2/sup 3/P/sub 0/ state from the close-lying states 2/sup 3/P/sub 2/, 2/sup 1/P/sub 1/, and 2/sup 3/P /sub 1/. In pD the corresponding transitions were found to be more than an order of magnitude broader. To a large extent the results for pH support the meson exchange model. (15 refs).

  14. High resolution multiplexed functional imaging in live embryos (Conference Presentation)

    Science.gov (United States)

    Xu, Dongli; Zhou, Weibin; Peng, Leilei

    2017-02-01

    Fourier multiplexed fluorescence lifetime imaging (FmFLIM) scanning laser optical tomography (FmFLIM-SLOT) combines FmFLIM and Scanning laser optical tomography (SLOT) to perform multiplexed 3D FLIM imaging of live embryos. The system had demonstrate multiplexed functional imaging of zebrafish embryos genetically express Foster Resonant Energy Transfer (FRET) sensors. However, previous system has a 20 micron resolution because the focused Gaussian beam diverges quickly from the focused plane, makes it difficult to achieve high resolution imaging over a long projection depth. Here, we present a high-resolution FmFLIM-SLOT system with achromatic Bessel beam, which achieves 3 micron resolution in 3D deep tissue imaging. In Bessel-FmFLIM-SLOT, multiple laser excitation lines are firstly intensity modulated by a Michelson interferometer with a spinning polygon mirror optical delay line, which enables Fourier multiplexed multi-channel lifetime measurements. Then, a spatial light modulator and a prism are used to transform the modulated Gaussian laser beam to an achromatic Bessel beam. The achromatic Bessel beam scans across the whole specimen with equal angular intervals as sample rotated. After tomography reconstruction and the frequency domain lifetime analysis method, both the 3D intensity and lifetime image of multiple excitation-emission can be obtained. Using Bessel-FmFLIM-SLOT system, we performed cellular-resolution FLIM tomography imaging of live zebrafish embryo. Genetically expressed FRET sensors in these embryo will allow non-invasive observation of multiple biochemical processes in vivo.

  15. Development of a metallic magnetic calorimeter for high resolution spectroscopy

    International Nuclear Information System (INIS)

    Linck, M.

    2007-01-01

    In this thesis the development of a metallic magnetic calorimeter for high resolution detection of single x-ray quanta is described. The detector consists of an X-ray absorber and a paramagnetic temperature sensor. The raise in temperature of the paramagnetic sensor due to the absorption of a single X-ray is measured by the change in magnetization of the sensor using a low-noise SQUID magnetometer. The thermodynamic properties of the detector can be described by a theoretical model based on a mean field approximation. This allows for an optimization of the detector design with respect to signal size. The maximal archivable energy resolution is limited by thermodynamic energy fluctuations between absorber, heat bath and thermometer. An interesting field of application for a metallic magnetic calorimeter is X-ray astronomy and the investigation of X-ray emitting objects. Through high-resolution X-ray spectroscopy it is possible to obtain information about physical processes of even far distant objects. The magnetic calorimeter that was developed in this thesis has a metallic absorber with a quantum efficiency of 98% at 6 keV. The energy resolution of the magnetic calorimeter is EFWHM=2.7 eV at 5.9 keV. The deviation of the detector response from a linear behavior of the detector is only 0.8% at 5.9 keV. (orig.)

  16. High resolution tsunami inversion for 2010 Chile earthquake

    Directory of Open Access Journals (Sweden)

    T.-R. Wu

    2011-12-01

    Full Text Available We investigate the feasibility of inverting high-resolution vertical seafloor displacement from tsunami waveforms. An inversion method named "SUTIM" (small unit tsunami inversion method is developed to meet this goal. In addition to utilizing the conventional least-square inversion, this paper also enhances the inversion resolution by Grid-Shifting method. A smooth constraint is adopted to gain stability. After a series of validation and performance tests, SUTIM is used to study the 2010 Chile earthquake. Based upon data quality and azimuthal distribution, we select tsunami waveforms from 6 GLOSS stations and 1 DART buoy record. In total, 157 sub-faults are utilized for the high-resolution inversion. The resolution reaches 10 sub-faults per wavelength. The result is compared with the distribution of the aftershocks and waveforms at each gauge location with very good agreement. The inversion result shows that the source profile features a non-uniform distribution of the seafloor displacement. The highly elevated vertical seafloor is mainly concentrated in two areas: one is located in the northern part of the epicentre, between 34° S and 36° S; the other is in the southern part, between 37° S and 38° S.

  17. Exogenous lipoid pneumonia: high-resolution CT findings

    International Nuclear Information System (INIS)

    Lee, J.S.; Song, K.S.; Lim, T.H.; Im, J.G.; Seo, J.B.

    1999-01-01

    The aim of this study was to assess high-resolution computed tomography (HRCT) findings of exogenous lipoid pneumonia. High-resolution computed tomography was obtained in 25 patients with proven exogenous lipoid pneumonia resulting from aspiration of squalene (derived from shark liver oil). Diagnosis was based on biopsy (n = 9), bronchoalveolar lavage (n = 8), or sputum cytology and clinical findings (n = 8). The clinical history of taking squalene was confirmed in all patients. The CT findings were classified into three patterns: diffuse ground-glass opacity, consolidation, and interstitial abnormalities. Distribution of the abnormalities, duration of taking squalene, predisposing factors for aspiration, and route of administration were analyzed. Ten patients showed diffuse ground-glass opacity pattern. Seven of 10 patients had predisposing conditions such as unconsciousness, pharyngeal dysmotility, or motor disturbances, and 6 patients had a recent history of taking large amount of squalene through nasal route. Seven patients who had consolidation pattern had a history of taking squalene for several months and did not have any predisposing factor. All of the 5 patients who had a pattern of interstitial abnormalities had a history of taking squalene longer than 1 year and showed segmental distribution of interstitial thickening with interposing ground-glass opacities. Three patients simultaneously had two different patterns at different lobes of the lung. The HRCT findings of lipoid pneumonia are ground-glass opacities, consolidation, and interstitial abnormalities. These HRCT findings with appropriate inquiries could be useful for diagnosis of exogeneous lipoid pneumonia. (orig.)

  18. Quantitative analysis of cholesteatoma using high resolution computed tomography

    International Nuclear Information System (INIS)

    Kikuchi, Shigeru; Yamasoba, Tatsuya; Iinuma, Toshitaka.

    1992-01-01

    Seventy-three cases of adult cholesteatoma, including 52 cases of pars flaccida type cholesteatoma and 21 of pars tensa type cholesteatoma, were examined using high resolution computed tomography, in both axial (lateral semicircular canal plane) and coronal sections (cochlear, vestibular and antral plane). These cases were classified into two subtypes according to the presence of extension of cholesteatoma into the antrum. Sixty cases of chronic otitis media with central perforation (COM) were also examined as controls. Various locations of the middle ear cavity were measured in terms of size in comparison with pars flaccida type cholesteatoma, pars tensa type cholesteatoma and COM. The width of the attic was significantly larger in both pars flaccida type and pars tensa type cholesteatoma than in COM. With pars flaccida type cholesteatoma there was a significantly larger distance between the malleus and lateral wall of the attic than with COM. In contrast, the distance between the malleus and medial wall of the attic was significantly larger with pars tensa type cholesteatoma than with COM. With cholesteatoma extending into the antrum, regardless of the type of cholesteatoma, there were significantly larger distances than with COM at the following sites: the width and height of the aditus ad antrum, and the width, height and anterior-posterior diameter of the antrum. However, these distances were not significantly different between cholesteatoma without extension into the antrum and COM. The hitherto demonstrated qualitative impressions of bone destruction in cholesteatoma were quantitatively verified in detail using high resolution computed tomography. (author)

  19. Characterization of ceramic archaeological by high resolution X ray microtomography

    International Nuclear Information System (INIS)

    Machado, Alessandra C.; Freitas, Renato; Calza, Cristiane F.; Lopes, Ricardo T.; Lima, Inaya; Carvalho, Daniele D.; Gaspar, Maria D.

    2013-01-01

    Characterization of ceramic fragments is a very important area of research in art and archeometry area because it enables a greater understanding of how ancient civilizations behave and what were their traditions and customs. Petrography and chemical analyses are commonly used, but these techniques are destructive, which is not interesting for this type of sample. Through the exchange of multidisciplinary scientific knowledge and new partnerships, high resolution X-ray microtomography has been introduced in archaeological area as a great possibility of 3D inspection in a non-destructive way. The goal of this work is to investigate the internal microstructures of four samples of archeological ceramic, from the Archaeological Site of Macacu - RJ. The X-ray microtomography were performed in a high resolution setup, and can be used to infer the nature of organic temper even with all plant remains completely burnt out during the firing process and also to ensure the homogeneity of samples envisaged for geochemical analyses, especially with respect to the distribution of chemically diverse fabric compounds. In this way this study intends to contribute to our understanding of the archaeological and historical formations of this region. (author)

  20. Characterization of ceramic archaeological by high resolution X ray microtomography

    Energy Technology Data Exchange (ETDEWEB)

    Machado, Alessandra C.; Freitas, Renato; Calza, Cristiane F.; Lopes, Ricardo T.; Lima, Inaya, E-mail: alecastro@lin.ufrj.br [Coordenacao dos Programas de Pos-Graduacao de Engenharia (COPPE/UFRJ), Rio de Janeiro, RJ (Brazil). Lab. de Instrumentacao Nuclear; Carvalho, Daniele D.; Gaspar, Maria D. [Museu Nacional (MN/UFRJ), RJ (Brazil). Centro de Tecnologia

    2013-07-01

    Characterization of ceramic fragments is a very important area of research in art and archeometry area because it enables a greater understanding of how ancient civilizations behave and what were their traditions and customs. Petrography and chemical analyses are commonly used, but these techniques are destructive, which is not interesting for this type of sample. Through the exchange of multidisciplinary scientific knowledge and new partnerships, high resolution X-ray microtomography has been introduced in archaeological area as a great possibility of 3D inspection in a non-destructive way. The goal of this work is to investigate the internal microstructures of four samples of archeological ceramic, from the Archaeological Site of Macacu - RJ. The X-ray microtomography were performed in a high resolution setup, and can be used to infer the nature of organic temper even with all plant remains completely burnt out during the firing process and also to ensure the homogeneity of samples envisaged for geochemical analyses, especially with respect to the distribution of chemically diverse fabric compounds. In this way this study intends to contribute to our understanding of the archaeological and historical formations of this region. (author)

  1. MR-Venography Using High Resolution True-FISP

    Energy Technology Data Exchange (ETDEWEB)

    Spuentrup, E. [Technische Hochschule Aachen (Germany). Klinik fuer Radiologische Diagnostik; Beth Israel Deaconess Medical Center, Boston, MA (United States). Dept. of Medicine; Harvard Medical School, Boston, MA (United States); Buecker, A.; Guenther, R.W. [Technische Hochschule Aachen (Germany). Klinik fuer Radiologische Diagnostik; Stuber, M. [Beth Israel Deaconess Medical Center, Boston, MA (United States). Dept. of Medicine; Harvard Medical School, Boston, MA (United States); Philips Med. Syst., Best (Netherlands)

    2001-08-01

    A new fast MR-venography approach using a high resolution True-FISP imaging sequence was investigated in 20 patients suffering from 23 deep vein thromboses. Diagnosis was proven by X-ray venography, CT or ultrasound examination. The presented technique allowed for clear thrombus visualization with a high contrast to the surrounding blood pool even in calf veins. Acquisition time was less than 10 minutes for imaging the pelvis and the legs. No contrast media was needed. The presented high resolution True-FISP MR-veography is a promising non-invasive, fast MR-venography approach for detection of deep venous thrombosis. (orig.) [German] Eine neue schnelle, oertlich hochaufgeloeste MR-Phlebographietechnik mit einer axialen True-FISP Bildgebungssequenz wurde an 20 Patienten mit 23 nach-gewiesenen tiefen Beinvenenthrombosen untersucht. Die Befunde wurden mit einer konventionellen Roentgenphlebographie, einer CT oder einer Sonographie gesichert. Die vorgestellte Technik erlaubte in allen Faellen eine Thrombusdarstellung mit hohem Kontrast zum umgebenden venoesen Blut, wobei aufgrund der hohen Ortsaufloesung auch die Unterschenkelvenen beurteilt werden konnten. Die Datenaufnahmezeit zur Untersuchung des Beckens und der Beine betrug weniger als 10 Minuten. Kontrastmittel wurde nicht benoetigt. Die vorgestellte MR-Phlebographietechnik unter Verwendung einer oertlich hochauf-geloesten True-FISP Sequenz ist eine neue, vielversprechende, nicht-invasive Technik zur Diagnostik der tiefen Bein- und Beckenvenenthrombose. (orig.)

  2. High-resolution electron microscopy of advanced materials

    Energy Technology Data Exchange (ETDEWEB)

    Mitchell, T.E.; Kung, H.H.; Sickafus, K.E.; Gray, G.T. III; Field, R.D.; Smith, J.F. [Los Alamos National Lab., NM (United States). Materials Science and Technology Div.

    1997-11-01

    This final report chronicles a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The High-Resolution Electron Microscopy Facility has doubled in size and tripled in quality since the beginning of the three-year period. The facility now includes a field-emission scanning electron microscope, a 100 kV field-emission scanning transmission electron microscope (FE-STEM), a 300 kV field-emission high-resolution transmission electron microscope (FE-HRTEM), and a 300 kV analytical transmission electron microscope. A new orientation imaging microscope is being installed. X-ray energy dispersive spectrometers for chemical analysis are available on all four microscopes; parallel electron energy loss spectrometers are operational on the FE-STEM and FE-HRTEM. These systems enable evaluation of local atomic bonding, as well as chemical composition in nanometer-scale regions. The FE-HRTEM has a point-to-point resolution of 1.6 {angstrom}, but the resolution can be pushed to its information limit of 1 {angstrom} by computer reconstruction of a focal series of images. HRTEM has been used to image the atomic structure of defects such as dislocations, grain boundaries, and interfaces in a variety of materials from superconductors and ferroelectrics to structural ceramics and intermetallics.

  3. Pulmonary leukemic involvement: high-resolution computed tomography evaluation

    International Nuclear Information System (INIS)

    Oliveira, Ana Paola de; Marchiori, Edson; Souza Junior, Arthur Soares

    2004-01-01

    Objective: To evaluate the role of high-resolution computed tomography (HRCT) in patients with leukemia and pulmonary symptoms, to establish the main patterns and to correlate them with the etiology. Materials and Methods: This is a retrospective study of the HRCT of 15 patients with leukemia and pulmonary symptoms. The examinations were performed using a spatial high-resolution protocol and were analyzed by two independent radiologists. Results: The main HRCT patterns found were ground-glass opacity (n=11), consolidation (n=9), airspace nodules (n=3), septal thickening (n=3), tree-in-bud pattern (n=3), and pleural effusion (n=3). Pulmonary infection was the most common finding seen in 12 patients: bacterial pneumonia (n=6), fungal infection (n = 4), pulmonary tuberculosis (n=1) and viral infection (n=1). Leukemic pleural infiltration (n=1), lymphoma (n=1) and pulmonary hemorrhage (n=1) were detected in the other three patients. Conclusion: HRCT is an important tool that may suggest the cause of lung involvement, its extension and in some cases to guide invasive procedures in patients with leukemia. (author)

  4. Peculiar velocity effects in high-resolution microwave background experiments

    International Nuclear Information System (INIS)

    Challinor, Anthony; Leeuwen, Floor van

    2002-01-01

    We investigate the impact of peculiar velocity effects due to the motion of the solar system relative to the cosmic microwave background (CMB) on high resolution CMB experiments. It is well known that on the largest angular scales the combined effects of Doppler shifts and aberration are important; the lowest Legendre multipoles of total intensity receive power from the large CMB monopole in transforming from the CMB frame. On small angular scales aberration dominates and is shown here to lead to significant distortions of the total intensity and polarization multipoles in transforming from the rest frame of the CMB to the frame of the solar system. We provide convenient analytic results for the distortions as series expansions in the relative velocity of the two frames, but at the highest resolutions a numerical quadrature is required. Although many of the high resolution multipoles themselves are severely distorted by the frame transformations, we show that their statistical properties distort by only an insignificant amount. Therefore, the cosmological parameter estimation is insensitive to the transformation from the CMB frame (where theoretical predictions are calculated) to the rest frame of the experiment

  5. Image thresholding in the high resolution target movement monitor

    Science.gov (United States)

    Moss, Randy H.; Watkins, Steve E.; Jones, Tristan H.; Apel, Derek B.; Bairineni, Deepti

    2009-03-01

    Image thresholding in the High Resolution Target Movement Monitor (HRTMM) is examined. The HRTMM was developed at the Missouri University of Science and Technology to detect and measure wall movements in underground mines to help reduce fatality and injury rates. The system detects the movement of a target with sub-millimeter accuracy based on the images of one or more laser dots projected on the target and viewed by a high-resolution camera. The relative position of the centroid of the laser dot (determined by software using thresholding concepts) in the images is the key factor in detecting the target movement. Prior versions of the HRTMM set the image threshold based on a manual, visual examination of the images. This work systematically examines the effect of varying threshold on the calculated centroid position and describes an algorithm for determining a threshold setting. First, the thresholding effects on the centroid position are determined for a stationary target. Plots of the centroid positions as a function of varying thresholds are obtained to identify clusters of thresholds for which the centroid position does not change for stationary targets. Second, the target is moved away from the camera in sub-millimeter increments and several images are obtained at each position and analyzed as a function of centroid position, target movement and varying threshold values. With this approach, the HRTMM can accommodate images in batch mode without the need for manual intervention. The capability for the HRTMM to provide automated, continuous monitoring of wall movement is enhanced.

  6. A high resolution pneumatic stepping actuator for harsh reactor environments

    Science.gov (United States)

    Tippetts, Thomas B.; Evans, Paul S.; Riffle, George K.

    1993-01-01

    A reactivity control actuator for a high-power density nuclear propulsion reactor must be installed in close proximity to the reactor core. The energy input from radiation to the actuator structure could exceed hundreds of W/cc unless low-cross section, low-absorptivity materials are chosen. Also, for post-test handling and subsequent storage, materials should not be used that are activated into long half-life isotopes. Pneumatic actuators can be constructed from various reactor-compatible materials, but conventional pneumatic piston actuators generally lack the stiffness required for high resolution reactivity control unless electrical position sensors and compensated electronic control systems are used. To overcome these limitations, a pneumatic actuator is under development that positions an output shaft in response to a series of pneumatic pulses, comprising a pneumatic analog of an electrical stepping motor. The pneumatic pulses are generated remotely, beyond the strong radiation environment, and transmitted to the actuator through tubing. The mechanically simple actuator uses a nutating gear harmonic drive to convert motion of small pistons directly to high-resolution angular motion of the output shaft. The digital nature of this actuator is suitable for various reactor control algorithms but is especially compatible with the three bean salad algorithm discussed by Ball et al. (1991).

  7. Evaluation of a high resolution silicon PET insert module

    Energy Technology Data Exchange (ETDEWEB)

    Grkovski, Milan, E-mail: milan.grkovski@ijs.si [Jožef Stefan Institute, Ljubljana (Slovenia); Memorial Sloan Kettering Cancer Center, New York, NY (United States); Brzezinski, Karol [IFIC/CSIC, Valencia (Spain); Cindro, Vladimir [Jožef Stefan Institute, Ljubljana (Slovenia); Clinthorne, Neal H. [University of Michigan, Ann Arbor, MI (United States); Kagan, Harris [Ohio State University, Columbus, OH (United States); Lacasta, Carlos [IFIC/CSIC, Valencia (Spain); Mikuž, Marko [Jožef Stefan Institute, Ljubljana (Slovenia); Solaz, Carles [IFIC/CSIC, Valencia (Spain); Studen, Andrej [Jožef Stefan Institute, Ljubljana (Slovenia); Weilhammer, Peter [Ohio State University, Columbus, OH (United States); Žontar, Dejan [Jožef Stefan Institute, Ljubljana (Slovenia)

    2015-07-11

    Conventional PET systems can be augmented with additional detectors placed in close proximity of the region of interest. We developed a high resolution PET insert module to evaluate the added benefit of such a combination. The insert module consists of two back-to-back 1 mm thick silicon sensors, each segmented into 1040 1 mm{sup 2} pads arranged in a 40 by 26 array. A set of 16 VATAGP7.1 ASICs and a custom assembled data acquisition board were used to read out the signal from the insert module. Data were acquired in slice (2D) geometry with a Jaszczak phantom (rod diameters of 1.2–4.8 mm) filled with {sup 18}F-FDG and the images were reconstructed with ML-EM method. Both data with full and limited angular coverage from the insert module were considered and three types of coincidence events were combined. The ratio of high-resolution data that substantially improves quality of the reconstructed image for the region near the surface of the insert module was estimated to be about 4%. Results from our previous studies suggest that such ratio could be achieved at a moderate technological expense by using an equivalent of two insert modules (an effective sensor thickness of 4 mm)

  8. The development of high resolution silicon x-ray microcalorimeters

    Science.gov (United States)

    Porter, F. S.; Kelley, R. L.; Kilbourne, C. A.

    2005-12-01

    Recently we have produced x-ray microcalorimeters with resolving powers approaching 2000 at 5.9 keV using a spare XRS microcalorimeter array. We attached 400 um square, 8 um thick HgTe absorbers using a variety of attachment methods to an XRS array and ran the detector array at temperatures between 40 and 60 mK. The best results were for absorbers attached using the standard XRS absorber-pixel thermal isolation scheme utilizing SU8 polymer tubes. In this scenario we achieved a resolution of 3.2 eV FWHM at 5.9 keV. Substituting a silicon spacer for the SU8 tubes also yielded sub-4eV results. In contrast, absorbers attached directly to the thermistor produced significant position dependence and thus degraded resolution. Finally, we tested standard 640um-square XRS detectors at reduced bias power at 50mK and achieved a resolution of 3.7eV, a 50% improvement over the XRS flight instrument. Implanted silicon microcalorimeters are a mature flight-qualified technology that still has a substantial phase space for future development. We will discuss these new high resolution results, the various absorber attachment schemes, planned future improvements, and, finally, their relevance to future high resolution x-ray spectrometers including Constellation-X.

  9. High resolution muon computed tomography at neutrino beam facilities

    International Nuclear Information System (INIS)

    Suerfu, B.; Tully, C.G.

    2016-01-01

    X-ray computed tomography (CT) has an indispensable role in constructing 3D images of objects made from light materials. However, limited by absorption coefficients, X-rays cannot deeply penetrate materials such as copper and lead. Here we show via simulation that muon beams can provide high resolution tomographic images of dense objects and of structures within the interior of dense objects. The effects of resolution broadening from multiple scattering diminish with increasing muon momentum. As the momentum of the muon increases, the contrast of the image goes down and therefore requires higher resolution in the muon spectrometer to resolve the image. The variance of the measured muon momentum reaches a minimum and then increases with increasing muon momentum. The impact of the increase in variance is to require a higher integrated muon flux to reduce fluctuations. The flux requirements and level of contrast needed for high resolution muon computed tomography are well matched to the muons produced in the pion decay pipe at a neutrino beam facility and what can be achieved for momentum resolution in a muon spectrometer. Such an imaging system can be applied in archaeology, art history, engineering, material identification and whenever there is a need to image inside a transportable object constructed of dense materials

  10. An original acquisition chain for the TOHR High Resolution Tomograph

    International Nuclear Information System (INIS)

    Pinot, Laurent

    1999-01-01

    The framework of this work is part of a new approach of emission tomography adapted to small animals. The principle of our tomographic system TOHR (French acronym for High Resolution Tomograph) is based on the use of large solid angle and high resolution focusing collimators each mounted in front of a detection module of high efficiency. With a first-generation acquisition chain we were able to characterize TOHR, however, to take fully advantage of the TOHR possibilities, a completely new acquisition scheme had to be designed. This system, being the main topic of this work, makes use of temporal information. The detection of a particle that entered the detector is translated into temporal logical signals. These signals pass into a time coding circuitry and the coded results are transferred in a digital processor. According to the initial terms of delivery, the developed acquisition chain steers the detection of events dependent on the deposited energy and time of arrival. The latter is done by coincidence measurements. All elements are mounted on a special board included into a PC unit and a dedicated program controls the whole system. First experiments showed up the interest of the new acquisition unit for other application in physics or medical imaging

  11. High resolution tsunami inversion for 2010 Chile earthquake

    Science.gov (United States)

    Wu, T.-R.; Ho, T.-C.

    2011-12-01

    We investigate the feasibility of inverting high-resolution vertical seafloor displacement from tsunami waveforms. An inversion method named "SUTIM" (small unit tsunami inversion method) is developed to meet this goal. In addition to utilizing the conventional least-square inversion, this paper also enhances the inversion resolution by Grid-Shifting method. A smooth constraint is adopted to gain stability. After a series of validation and performance tests, SUTIM is used to study the 2010 Chile earthquake. Based upon data quality and azimuthal distribution, we select tsunami waveforms from 6 GLOSS stations and 1 DART buoy record. In total, 157 sub-faults are utilized for the high-resolution inversion. The resolution reaches 10 sub-faults per wavelength. The result is compared with the distribution of the aftershocks and waveforms at each gauge location with very good agreement. The inversion result shows that the source profile features a non-uniform distribution of the seafloor displacement. The highly elevated vertical seafloor is mainly concentrated in two areas: one is located in the northern part of the epicentre, between 34° S and 36° S; the other is in the southern part, between 37° S and 38° S.

  12. High resolution and simultaneous monitoring of airborne radionuclides

    International Nuclear Information System (INIS)

    Abe, T.; Yamaguchi, Y.; Muguntha Manikandan, N.; Komura, K.

    2005-01-01

    By using 11 extremely low background Ge detectors at Ogoya Underground Laboratory, it became possible to investigate temporal variations of airborne 212 Pb (T 1/2 =10.6 h) along with 210 Pb and 7 Be with order of magnitude higher time resolution. Then, we have measured airborne nuclides at three monitoring points, (1) roof of our laboratory (LLRL; 40 m ASL), (2) Shinshiku Plateau (640 m ASL) located about 8 km from LLRL as a comparison of vertical distribution, and (3) Hegura Island (10 m ASL) at about 50 km from Wajima located north of Noto Peninsula facing on the Sea of Japan (about 180 km to the north-northeast of LLRL), to investigate influence of Asian continent. Airborne nuclides were collected by high volume air samplers at intervals of a few hours at either two or three points simultaneously. In the same manner, high resolution monitoring was carried out also at the time of passage of typhoon and cold front. In this study, we observed drastic temporal variations of airborne radionuclides and correlations of multiple monitoring points. The results indicate that high resolution and simultaneous monitoring is very useful to understand dynamic state of variations of airborne nuclides due to short and long-term air-mass movement. (author)

  13. High-Resolution Electronics: Spontaneous Patterning of High-Resolution Electronics via Parallel Vacuum Ultraviolet (Adv. Mater. 31/2016).

    Science.gov (United States)

    Liu, Xuying; Kanehara, Masayuki; Liu, Chuan; Sakamoto, Kenji; Yasuda, Takeshi; Takeya, Jun; Minari, Takeo

    2016-08-01

    On page 6568, T. Minari and co-workers describe spontaneous patterning based on the parallel vacuum ultraviolet (PVUV) technique, enabling the homogeneous integration of complex, high-resolution electronic circuits, even on large-scale, flexible, transparent substrates. Irradiation of PVUV to the hydrophobic polymer surface precisely renders the selected surface into highly wettable regions with sharply defined boundaries, which spontaneously guides a metal nanoparticle ink into a series of circuit lines and gaps with the widths down to a resolution of 1 μm. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. High resolution modelling of extreme precipitation events in urban areas

    Science.gov (United States)

    Siemerink, Martijn; Volp, Nicolette; Schuurmans, Wytze; Deckers, Dave

    2015-04-01

    The present day society needs to adjust to the effects of climate change. More extreme weather conditions are expected, which can lead to longer periods of drought, but also to more extreme precipitation events. Urban water systems are not designed for such extreme events. Most sewer systems are not able to drain the excessive storm water, causing urban flooding. This leads to high economic damage. In order to take appropriate measures against extreme urban storms, detailed knowledge about the behaviour of the urban water system above and below the streets is required. To investigate the behaviour of urban water systems during extreme precipitation events new assessment tools are necessary. These tools should provide a detailed and integral description of the flow in the full domain of overland runoff, sewer flow, surface water flow and groundwater flow. We developed a new assessment tool, called 3Di, which provides detailed insight in the urban water system. This tool is based on a new numerical methodology that can accurately deal with the interaction between overland runoff, sewer flow and surface water flow. A one-dimensional model for the sewer system and open channel flow is fully coupled to a two-dimensional depth-averaged model that simulates the overland flow. The tool uses a subgrid-based approach in order to take high resolution information of the sewer system and of the terrain into account [1, 2]. The combination of using the high resolution information and the subgrid based approach results in an accurate and efficient modelling tool. It is now possible to simulate entire urban water systems using extreme high resolution (0.5m x 0.5m) terrain data in combination with a detailed sewer and surface water network representation. The new tool has been tested in several Dutch cities, such as Rotterdam, Amsterdam and The Hague. We will present the results of an extreme precipitation event in the city of Schiedam (The Netherlands). This city deals with

  15. High resolution remote sensing of water surface patterns

    Science.gov (United States)

    Woodget, A.; Visser, F.; Maddock, I.; Carbonneau, P.

    2012-12-01

    The assessment of in-stream habitat availability within fluvial environments in the UK traditionally includes the mapping of patterns which appear on the surface of the water, known as 'surface flow types' (SFTs). The UK's River Habitat Survey identifies ten key SFTs, including categories such as rippled flow, upwelling, broken standing waves and smooth flow. SFTs result from the interaction between the underlying channel morphology, water depth and velocity and reflect the local flow hydraulics. It has been shown that SFTs can be both biologically and hydraulically distinct. SFT mapping is usually conducted from the river banks where estimates of spatial coverage are made by eye. This approach is affected by user subjectivity and inaccuracies in the spatial extent of mapped units. Remote sensing and specifically the recent developments in unmanned aerial systems (UAS) may now offer an alternative approach for SFT mapping, with the capability for rapid and repeatable collection of very high resolution imagery from low altitudes, under bespoke flight conditions. This PhD research is aimed at investigating the mapping of SFTs using high resolution optical imagery (less than 10cm) collected from a helicopter-based UAS flown at low altitudes (less than 100m). This paper presents the initial findings from a series of structured experiments on the River Arrow, a small lowland river in Warwickshire, UK. These experiments investigate the potential for mapping SFTs from still and video imagery of different spatial resolutions collected at different flying altitudes and from different viewing angles (i.e. vertical and oblique). Imagery is processed using 3D mosaicking software to create orthophotos and digital elevation models (DEM). The types of image analysis which are tested include a simple, manual visual assessment undertaken in a GIS environment, based on the high resolution optical imagery. In addition, an object-based image analysis approach which makes use of the

  16. A PICTORIAL PRESENTATION OF ESOPHAGEAL HIGH RESOLUTION MANOMETRY CURRENT PARAMETERS.

    Science.gov (United States)

    Lafraia, Fernanda M; Herbella, Fernando A M; Kalluf, Julia R; Patti, Marco G

    2017-01-01

    High resolution manometry is the current technology used to the study of esophageal motility and is replacing conventional manometry in important centers for esophageal motility with parameters used on esophageal motility, following the Chicago Classification. This classification unifies high resolution manometry interpretation and classifies esophageal disorders. This review shows, in a pictorial presentation, the new parameters established by the Chicago Classification, version 3.0, aimed to allow an easy comprehension and interpretation of high resolution manometry. Esophageal manometries performed by the authors were reviewed to select illustrative tracings representing Chicago Classification parameters. The parameters are: Esophagogastric Morphology, that classifies this junction according to its physiology and anatomy; Integrated Relaxation Pressure, that measures the lower esophageal sphincter relaxation; Distal Contractile Integral, that evaluates the contraction vigor of each wave; and, Distal Latency, that measures the peristalsis velocity from the beginning of the swallow to the epiphrenic ampulla. Clinical applications of these new concepts is still under evaluation. Mostrar, de forma pictórica, os novos parâmetros compilados na versão 3.0 da Classificação de Chicago, buscando facilitar a compreensão e interpretação da manometria de alta resolução. Foram revistas as manometrias da casuística dos autores e selecionados os traçados representativos dos parâmetros da Classificação de Chicago. Entre os parâmetros apresentados foram considerados a Morfologia da Transição Gastroesofágica, que classifica o segmento de acordo com sua fisiologia e anatomia; a Integral da Pressão de Relaxamento, que mede o relaxamento do esfíncter esofagiano inferior; a Integral Contrátil Distal, que avalia o vigor contrátil da onda peristáltica; e, a Latência Distal, que mede o tempo da peristalse, desde o início da deglutição até a ampola epifr

  17. Quantifying and containing the curse of high resolution coronal imaging

    Directory of Open Access Journals (Sweden)

    V. Delouille

    2008-10-01

    Full Text Available Future missions such as Solar Orbiter (SO, InterHelioprobe, or Solar Probe aim at approaching the Sun closer than ever before, with on board some high resolution imagers (HRI having a subsecond cadence and a pixel area of about (80 km2 at the Sun during perihelion. In order to guarantee their scientific success, it is necessary to evaluate if the photon counts available at these resolution and cadence will provide a sufficient signal-to-noise ratio (SNR. For example, if the inhomogeneities in the Quiet Sun emission prevail at higher resolution, one may hope to locally have more photon counts than in the case of a uniform source. It is relevant to quantify how inhomogeneous the quiet corona will be for a pixel pitch that is about 20 times smaller than in the case of SoHO/EIT, and 5 times smaller than TRACE. We perform a first step in this direction by analyzing and characterizing the spatial intermittency of Quiet Sun images thanks to a multifractal analysis. We identify the parameters that specify the scale-invariance behavior. This identification allows next to select a family of multifractal processes, namely the Compound Poisson Cascades, that can synthesize artificial images having some of the scale-invariance properties observed on the recorded images. The prevalence of self-similarity in Quiet Sun coronal images makes it relevant to study the ratio between the SNR present at SoHO/EIT images and in coarsened images. SoHO/EIT images thus play the role of "high resolution" images, whereas the "low-resolution" coarsened images are rebinned so as to simulate a smaller angular resolution and/or a larger distance to the Sun. For a fixed difference in angular resolution and in Spacecraft-Sun distance, we determine the proportion of pixels having a SNR preserved at high resolution given a particular increase in effective area. If scale-invariance continues to prevail at smaller scales, the conclusion reached with SoHO/EIT images can be transposed

  18. Investigation of the Behavior of Ethylene Molecular Films Using High Resolution Adsorption Isotherms and Neutron Scattering

    International Nuclear Information System (INIS)

    Barbour, Andi M.; Telling, Mark T.; Larese, John Z.

    2010-01-01

    The wetting behavior of ethylene adsorbed on MgO(100) was investigated from 83-135 K using high resolution volumetric adsorption isotherms. The results are compared to ethylene adsorption on graphite, a prototype adsorption system, in an effort to gain further insight into the forces that drive the observed film growth. Layering transitions for ethylene on MgO(100) are observed below the bulk triple point of ethylene (T = 104.0 K). The formation of three discrete adlayers is observed on the MgO(100) surface; onset of the second and third layers occurs at 79.2 ± 1.3 K and 98.3 ± 0.9 K, respectively. Thermodynamic quantities such as differential enthalpy and entropy, heat of adsorption, and isosteric heat of adsorption are determined and compared to the previously published values for ethylene on graphite. The average area occupied by a ethylene molecule on MgO(100) is 22.6 ± 1.1 (angstrom) 2 molecule -1 . The locations of two phase transitions are identified (i.e., layer critical temperatures at T c2 (n=1) at 108.6 ± 1.7 K and T c2 (n=2) at 116.5 ± 1.2 K) and a phase diagram is proposed. Preliminary neutron diffraction measurements reveal evidence of a monolayer solid with a lattice constant of ∼4.2 (angstrom). High resolution INS measurements show that the onset to dynamical motion and monolayer melting take place at 35 K and 65 K, respectively. The data reported here exhibit a striking similarity to ethylene on graphite which suggests that molecule-molecule interactions play an important role in determining the physical properties and growth of molecularly thin ethylene films.

  19. A high-resolution Fourier Transform Spectrometer for planetary spectroscopy

    Science.gov (United States)

    Cruikshank, D. P.; Sinton, W. M.

    1973-01-01

    The employment of a high-resolution Fourier Transform Spectrometer (FTS) is described for planetary and other astronomical spectroscopy in conjunction with the 88-inch telescope at Mauna Kea Observatory. The FTS system is designed for a broad range of uses, including double-beam laboratory spectroscopy, infrared gas chromatography, and nuclear magnetic resonance spectroscopy. The data system is well-suited to astronomical applications because of its great speed in acquiring and transforming data, and because of the enormous storage capability of the magnetic tape unit supplied with the system. The basic instrument is outlined 2nd some of the initial results from the first attempted use on the Mauna Kea 88-inch telescope are reported.

  20. High Resolution Energetic X-ray Imager (HREXI)

    Science.gov (United States)

    Grindlay, Jonathan

    We propose to design and build the first imaging hard X-ray detector system that incorporates 3D stacking of closely packed detector readouts in finely-spaced imaging arrays with their required data processing and control electronics. In virtually all imaging astronomical detectors, detector readout is done with flex connectors or connections that are not vertical but rather horizontal , requiring loss of focal plane area. For high resolution pixel detectors needed for high speed event-based X-ray imaging, from low energy applications (CMOS) with focusing X-ray telescopes, to hard X-ray applications with pixelated CZT for large area coded aperture telescopes, this new detector development offers great promise. We propose to extend our previous and current APRA supported ProtoEXIST program that has developed the first large area imaging CZT detectors and demonstrated their astrophysical capabilities on two successful balloon flight to a next generation High Resolution Energetic X-ray Imager (HREXI), which would incorporate microvia technology for the first time to connect the readout ASIC on each CZT crystal directly to its control and data processing system. This 3-dimensional stacking of detector and readout/control system means that large area (>2m2) imaging detector planes for a High Resolution Wide-field hard X-ray telescope can be built with initially greatly reduced detector gaps and ultimately with no gaps. This increases detector area, efficiency, and simplicity of detector integration. Thus higher sensitivity wide-field imagers will be possible at lower cost. HREXI will enable a post-Swift NASA mission such as the EREXS concept proposed to PCOS to be conducted as a future MIDEX mission. This mission would conduct a high resolution (<2 arcmin) , broad band (5 200 keV) hard X-ray survey of black holes on all scales with ~10X higher sensitivity than Swift. In the current era of Time Domain Astrophysics, such a survey capability, in conjunction with a n

  1. Unattented mode monitoring of high resolution gamma-ray spectra

    International Nuclear Information System (INIS)

    Smith, B.G.R.; Van Dyck, P.; Debraix, P.

    1991-01-01

    An Isotope Monitoring System (IMS) for unattended spectrum acquisition is described. This consists of a simple low cost flexible software package running on a Compaq 80386 and controlling up to 4 Canberra Packard System 100 multi-channel analyzer (MCA) cards. The IMS permits the independent configuration of each of the 4 MCA cards for different monitoring cycles and for different spectrum acquisition cycles each being based upon different trigger criteria. In this way IMS is able to automatically identify events, time tag them, and acquire and store valid spectra corresponding to those event. An additional feature of IMS permits to run a Multigroup Analysis (MGA) software package for the determination of plutonium isotopic compositions in batch mode. One particular application is discussed which consists of 4 high resolution gamma-ray detector systems connected together to the IMS for unattended spectrum acquisition. The off-line batch mode analysis of the spectra using MGA is also discussed

  2. High-resolution method for evolving complex interface networks

    Science.gov (United States)

    Pan, Shucheng; Hu, Xiangyu Y.; Adams, Nikolaus A.

    2018-04-01

    In this paper we describe a high-resolution transport formulation of the regional level-set approach for an improved prediction of the evolution of complex interface networks. The novelty of this method is twofold: (i) construction of local level sets and reconstruction of a global level set, (ii) local transport of the interface network by employing high-order spatial discretization schemes for improved representation of complex topologies. Various numerical test cases of multi-region flow problems, including triple-point advection, single vortex flow, mean curvature flow, normal driven flow, dry foam dynamics and shock-bubble interaction show that the method is accurate and suitable for a wide range of complex interface-network evolutions. Its overall computational cost is comparable to the Semi-Lagrangian regional level-set method while the prediction accuracy is significantly improved. The approach thus offers a viable alternative to previous interface-network level-set method.

  3. High resolution ultrasound and photoacoustic imaging of single cells

    Directory of Open Access Journals (Sweden)

    Eric M. Strohm

    2016-03-01

    Full Text Available High resolution ultrasound and photoacoustic images of stained neutrophils, lymphocytes and monocytes from a blood smear were acquired using a combined acoustic/photoacoustic microscope. Photoacoustic images were created using a pulsed 532 nm laser that was coupled to a single mode fiber to produce output wavelengths from 532 nm to 620 nm via stimulated Raman scattering. The excitation wavelength was selected using optical filters and focused onto the sample using a 20× objective. A 1000 MHz transducer was co-aligned with the laser spot and used for ultrasound and photoacoustic images, enabling micrometer resolution with both modalities. The different cell types could be easily identified due to variations in contrast within the acoustic and photoacoustic images. This technique provides a new way of probing leukocyte structure with potential applications towards detecting cellular abnormalities and diseased cells at the single cell level.

  4. HIGH-RESOLUTION ATMOSPHERIC ENSEMBLE MODELING AT SRNL

    Energy Technology Data Exchange (ETDEWEB)

    Buckley, R.; Werth, D.; Chiswell, S.; Etherton, B.

    2011-05-10

    The High-Resolution Mid-Atlantic Forecasting Ensemble (HME) is a federated effort to improve operational forecasts related to precipitation, convection and boundary layer evolution, and fire weather utilizing data and computing resources from a diverse group of cooperating institutions in order to create a mesoscale ensemble from independent members. Collaborating organizations involved in the project include universities, National Weather Service offices, and national laboratories, including the Savannah River National Laboratory (SRNL). The ensemble system is produced from an overlapping numerical weather prediction model domain and parameter subsets provided by each contributing member. The coordination, synthesis, and dissemination of the ensemble information are performed by the Renaissance Computing Institute (RENCI) at the University of North Carolina-Chapel Hill. This paper discusses background related to the HME effort, SRNL participation, and example results available from the RENCI website.

  5. Precision Viticulture from Multitemporal, Multispectral Very High Resolution Satellite Data

    Science.gov (United States)

    Kandylakis, Z.; Karantzalos, K.

    2016-06-01

    In order to exploit efficiently very high resolution satellite multispectral data for precision agriculture applications, validated methodologies should be established which link the observed reflectance spectra with certain crop/plant/fruit biophysical and biochemical quality parameters. To this end, based on concurrent satellite and field campaigns during the veraison period, satellite and in-situ data were collected, along with several grape samples, at specific locations during the harvesting period. These data were collected for a period of three years in two viticultural areas in Northern Greece. After the required data pre-processing, canopy reflectance observations, through the combination of several vegetation indices were correlated with the quantitative results from the grape/must analysis of grape sampling. Results appear quite promising, indicating that certain key quality parameters (like brix levels, total phenolic content, brix to total acidity, anthocyanin levels) which describe the oenological potential, phenolic composition and chromatic characteristics can be efficiently estimated from the satellite data.

  6. High-Resolution Characterization of UMo Alloy Microstructure

    Energy Technology Data Exchange (ETDEWEB)

    Devaraj, Arun [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Kovarik, Libor [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Jana, Saumyadeep [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Manandhar, Sandeep [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Arey, Bruce W. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-11-30

    This report highlights the capabilities and procedure for high-resolution characterization of UMo fuels in PNNL. Uranium-molybdenum (UMo) fuel processing steps, from casting to forming final fuel, directly affect the microstructure of the fuel, which in turn dictates the in-reactor performance of the fuel under irradiation. In order to understand the influence of processing on UMo microstructure, microstructure characterization techniques are necessary. Higher-resolution characterization techniques like transmission electron microscopy (TEM) and atom probe tomography (APT) are needed to interrogate the details of the microstructure. The findings from TEM and APT are also directly beneficial for developing predictive multiscale modeling tools that can predict the microstructure as a function of process parameters. This report provides background on focused-ion-beam–based TEM and APT sample preparation, TEM and APT analysis procedures, and the unique information achievable through such advanced characterization capabilities for UMo fuels, from a fuel fabrication capability viewpoint.

  7. THE HIGH-RESOLUTION INFRARED SPECTRUM OF HCl{sup +}

    Energy Technology Data Exchange (ETDEWEB)

    Doménech, J. L.; Herrero, V. J.; Tanarro, I. [Molecular Physics Department, Instituto de Estructura de la Materia (IEM-CSIC), Serrano 123, E-28006 Madrid (Spain); Drouin, B. J. [Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91109-8099 (United States); Cernicharo, J., E-mail: jl.domenech@csic.es [Molecular Astrophysics Group, Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC), Sor Juana Inés de la Cruz 3, Cantoblanco, E-28049 Madrid (Spain)

    2016-12-20

    The chloroniumyl cation, HCl{sup +}, has been recently identified in space from Herschel 's spectra. A joint analysis of extensive vis-UV spectroscopy emission data together with a few high-resolution and high-accuracy millimeter-wave data provided the necessary rest frequencies to support the astronomical identification. Nevertheless, the analysis did not include any infrared (IR) vibration–rotation data. Furthermore, with the end of the Herschel mission, IR observations from the ground may be one of the few available means to further study this ion in space. In this work, we provide a set of accurate rovibrational transition wavenumbers, as well as a new and improved global fit of vis-UV, IR, and millimeter-wave spectroscopy laboratory data, that will aid in future studies of this molecule.

  8. Measuring large-scale social networks with high resolution.

    Directory of Open Access Journals (Sweden)

    Arkadiusz Stopczynski

    Full Text Available This paper describes the deployment of a large-scale study designed to measure human interactions across a variety of communication channels, with high temporal resolution and spanning multiple years-the Copenhagen Networks Study. Specifically, we collect data on face-to-face interactions, telecommunication, social networks, location, and background information (personality, demographics, health, politics for a densely connected population of 1000 individuals, using state-of-the-art smartphones as social sensors. Here we provide an overview of the related work and describe the motivation and research agenda driving the study. Additionally, the paper details the data-types measured, and the technical infrastructure in terms of both backend and phone software, as well as an outline of the deployment procedures. We document the participant privacy procedures and their underlying principles. The paper is concluded with early results from data analysis, illustrating the importance of multi-channel high-resolution approach to data collection.

  9. Application of the Oslo method to high resolution gamma spectra

    Science.gov (United States)

    Simon, A.; Guttormsen, M.; Larsen, A. C.; Beausang, C. W.; Humby, P.

    2015-10-01

    Hauser-Feshbach statistical model is a widely used tool for calculation of the reaction cross section, in particular for astrophysical processes. The HF model requires as an input an optical potential, gamma-strength function (GSF) and level density (LD) to properly model the statistical properties of the nucleus. The Oslo method is a well established technique to extract GSFs and LDs from experimental data, typically used for gamma-spectra obtained with scintillation detectors. Here, the first application of the Oslo method to high-resolution data obtained using the Ge detectors of the STARLITER setup at TAMU is discussed. The GSFs and LDs extracted from (p,d) and (p,t) reactions on 152154 ,Sm targets will be presented.

  10. Highly sensitive high resolution Raman spectroscopy using resonant ionization methods

    International Nuclear Information System (INIS)

    Owyoung, A.; Esherick, P.

    1984-05-01

    In recent years, the introduction of stimulated Raman methods has offered orders of magnitude improvement in spectral resolving power for gas phase Raman studies. Nevertheless, the inherent weakness of the Raman process suggests the need for significantly more sensitive techniques in Raman spectroscopy. In this we describe a new approach to this problem. Our new technique, which we call ionization-detected stimulated Raman spectroscopy (IDSRS), combines high-resolution SRS with highly-sensitive resonant laser ionization to achieve an increase in sensitivity of over three orders of magnitude. The excitation/detection process involves three sequential steps: (1) population of a vibrationally excited state via stimulated Raman pumping; (2) selective ionization of the vibrationally excited molecule with a tunable uv source; and (3) collection of the ionized species at biased electrodes where they are detected as current in an external circuit

  11. High resolution computed tomography of the middle ear

    International Nuclear Information System (INIS)

    Ikeda, Katsuhisa; Sakurai, Tokio; Saijo, Shigeru; Kobayashi, Toshimitsu

    1983-01-01

    High resolution computed tomography was performed in 57 cases with various middle ear diseases (chronic otitis media, otitis media with effusion, acute otitis media and atelectasis). Although further improvement in detectability is necessary in order to discriminate each type of the soft tissue lesions, CT is the most useful method currently available in detecting the small structures and soft tissue lesions of the middle ear. In particular, the lesions at the tympanic isthmus and tympanic fold could very clearly be detected only by CT. In acute otitis media, lesions usually started in the attic and spread to the mastoid air cells. In otitis media with effusion, the soft tissue shadow was ovserved in the attic and mastoid air cell. CT is valuable in diagnosis, evaluation of the treatment and prognosis, and analysis of pathophysiology in the middle ear diseases. (author)

  12. High resolution X-ray tomography for stationary multiphase flows

    International Nuclear Information System (INIS)

    Schmitz, D.; Reinecke, N.; Petritsch, G.; Mewes, D.

    1998-01-01

    The high resolution which can be obtained by computer assisted tomography is used to investigate the liquid distribution and void fraction in random and structured packing. With a spatial resolution of 0.4x0.4mm 2 it is possible even to detect thin liquid films on structured packings. The experimental set-up consists of a custom-built second generation tomograph. The imaged object consists of a column filled with either a random ceramic packing of spheres or a structured metal packing. The liquid and void fraction distribution in random and structured packings with a quiescent gaseous phase is visualized. The water/air system is used. The liquid distributor consists of a perforated plate. The experimental hold-up values averaged over the column cross-section are in good agreement with empirical equations. (author)

  13. Fast iterative segmentation of high resolution medical images

    International Nuclear Information System (INIS)

    Hebert, T.J.

    1996-01-01

    Various applications in positron emission tomography (PET), single photon emission computed tomography (SPECT) and magnetic resonance imaging (MRI) require segmentation of 20 to 60 high resolution images of size 256x256 pixels in 3-9 seconds per image. This places particular constraints on the design of image segmentation algorithms. This paper examines the trade-offs in segmenting images based on fitting a density function to the pixel intensities using curve-fitting versus the maximum likelihood method. A quantized data representation is proposed and the EM algorithm for fitting a finite mixture density function to the quantized representation for an image is derived. A Monte Carlo evaluation of mean estimation error and classification error showed that the resulting quantized EM algorithm dramatically reduces the required computation time without loss of accuracy

  14. High resolution capacitance detection circuit for rotor micro-gyroscope

    Directory of Open Access Journals (Sweden)

    Ming-Yuan Ren

    2014-03-01

    Full Text Available Conventional methods for rotor position detection of micro-gyroscopes include common exciting electrodes (single frequency and common sensing electrodes (frequency multiplex, but they have encountered some problems. So we present a high resolution and low noise pick-off circuit for micro-gyroscopes which utilizes the time multiplex method. The detecting circuit adopts a continuous-time current sensing circuit for capacitance measurement, and its noise analysis of the charge amplifier is introduced. The equivalent output noise power spectral density of phase-sensitive demodulation is 120 nV/Hz1/2. Tests revealed that the whole circuitry has a relative capacitance resolution of 1 × 10−8.

  15. Automated high-resolution NMR with a sample changer

    International Nuclear Information System (INIS)

    Wade, C.G.; Johnson, R.D.; Philson, S.B.; Strouse, J.; McEnroe, F.J.

    1989-01-01

    Within the past two years, it has become possible to obtain high-resolution NMR spectra using automated commercial instrumentation. Software control of all spectrometer functions has reduced most of the tedious manual operations to typing a few computer commands or even making selections from a menu. Addition of an automatic sample changer is the next natural step in improving efficiency and sample throughput; it has a significant (and even unexpected) impact on how NMR laboratories are run and how it is taught. Such an instrument makes even sophisticated experiments routine, so that people with no previous exposure to NMR can run these experiments after a training session of an hour or less. This A/C Interface examines the impact of such instrumentation on both the academic and the industrial laboratory

  16. DETECTION OF BARCHAN DUNES IN HIGH RESOLUTION SATELLITE IMAGES

    Directory of Open Access Journals (Sweden)

    M. A. Azzaoui

    2016-06-01

    Full Text Available Barchan dunes are the fastest moving sand dunes in the desert. We developed a process to detect barchans dunes on High resolution satellite images. It consisted of three steps, we first enhanced the image using histogram equalization and noise reduction filters. Then, the second step proceeds to eliminate the parts of the image having a texture different from that of the barchans dunes. Using supervised learning, we tested a coarse to fine textural analysis based on Kolomogorov Smirnov test and Youden’s J-statistic on co-occurrence matrix. As an output we obtained a mask that we used in the next step to reduce the search area. In the third step we used a gliding window on the mask and check SURF features with SVM to get barchans dunes candidates. Detected barchans dunes were considered as the fusion of overlapping candidates. The results of this approach were very satisfying in processing time and precision.

  17. High resolution computed tomography of the post partum pituitary gland

    International Nuclear Information System (INIS)

    Hinshaw, D.B.; Hasso, A.N.; Thompson, J.R.; Davidson, B.J.

    1984-01-01

    Eight volunteer post partum female patients were examined with high resolution computed tomography during the week immediately after delivery. All patients received high dose (40-70 gm) intravenous iodine contrast administration. The scans were examined for pituitary gland height, shape and homogeneity. All of the patients had enlarged glands by the traditional standards (i.e. gland height of 8 mm or greater). The diaphragma sellae in every call bulged upward with a convex domed appearance. The glands were generally inhomogeneous. One gland had a 4 mm focal well defined area of decreased attenuation. Two patients who were studied again months later had glands which had returned to ''normal'' size. The enlarged, upwardly convex pituitary gland appears to be typical and normal for the recently post partum period. (orig.)

  18. High-resolution sonography in carpal tunnel syndrome

    International Nuclear Information System (INIS)

    Solbiati, L.; De Pra, L.; Rizzatto, G.; Derchi, L.E.

    1986-01-01

    Carpal tunnel syndrome, caused by the compression on the median nerve under the transverse carpal ligament, has multiple causes and clinical presentations. One hundred eighteen patients with carpal tunnel sydrome underwent high-resolution US which demonstrated unpalpable cystic masses in 25 patients (lobulated stalked synovial cysts in 19 and retrotendinous cysts in six, all confirmed at surgery), and diffuse thickening and decreased echogenicity of the tendon sheaths in 87 patients, suggesting tenosynovitis (confirmed at surgery in 64). In six patients simple encasement of muscle bellies in the carpal tunnel was shown. US can delineate the cause of carpal tunnel syndrome, suggest the need for surgery, and aid the surgeon in locating the lesion to be removed

  19. Feasibility tests of a high resolution sampling radial drift chamber

    International Nuclear Information System (INIS)

    Huth, J.

    1985-01-01

    The design concept and results of feasibility tests for a vertex detector intended for use in the TPC-PEP4/9 experiment are presented. The detector is based on a slow radial drift in dimethyl ether. High resolution localization of the avalanches at the sense wire is accomplished with nearby pickup wires and the utilization of waveform sampling electronics. The avalanche angular coordinate measurements, combined with knowledge of the electric field distribution and drift velocity permit reconstruction of the trajectory using essentially all track information. Measurements with a test chamber constructed to study characteristics of avalanche localization indicate that the recoverable track information in one centimeter of dimethyl ether at 1.5 atm is equivalent to 30 measurements of 40 μm accuracy. (orig.)

  20. Sparse Representation Denoising for Radar High Resolution Range Profiling

    Directory of Open Access Journals (Sweden)

    Min Li

    2014-01-01

    Full Text Available Radar high resolution range profile has attracted considerable attention in radar automatic target recognition. In practice, radar return is usually contaminated by noise, which results in profile distortion and recognition performance degradation. To deal with this problem, in this paper, a novel denoising method based on sparse representation is proposed to remove the Gaussian white additive noise. The return is sparsely described in the Fourier redundant dictionary and the denoising problem is described as a sparse representation model. Noise level of the return, which is crucial to the denoising performance but often unknown, is estimated by performing subspace method on the sliding subsequence correlation matrix. Sliding window process enables noise level estimation using only one observation sequence, not only guaranteeing estimation efficiency but also avoiding the influence of profile time-shift sensitivity. Experimental results show that the proposed method can effectively improve the signal-to-noise ratio of the return, leading to a high-quality profile.

  1. Neutron spin echo and high resolution inelastic spectroscopy

    International Nuclear Information System (INIS)

    Mezei, F.; Hungarian Academy of Sciences, Budapest. Central Research Inst. for Physics)

    1982-01-01

    The principles of neutrons spin echo (NSE) technique are considered. It is shown that the basis of NSE principle is a single step measurement of the change of the neutron velocity in the scattering process. The backscattering soectroscopy and the NSE techniques are compared. The NSF spectrometer is described. It is shown that 0.5 MeV energy resolution achieved in the NSE experiment is about 40 times superior to those achieved by the other techniques. The NSE technique has the unique feature that provides high resolution in neutron energy change independently of the monochromatization of the beam. The NSE instrument not only covers a wider dynamic range on a pulsed source that on a continuous one, but also collects data more efficiently

  2. High-resolution electron microscopy and its applications.

    Science.gov (United States)

    Li, F H

    1987-12-01

    A review of research on high-resolution electron microscopy (HREM) carried out at the Institute of Physics, the Chinese Academy of Sciences, is presented. Apart from the direct observation of crystal and quasicrystal defects for some alloys, oxides, minerals, etc., and the structure determination for some minute crystals, an approximate image-contrast theory named pseudo-weak-phase object approximation (PWPOA), which shows the image contrast change with crystal thickness, is described. Within the framework of PWPOA, the image contrast of lithium ions in the crystal of R-Li2Ti3O7 has been observed. The usefulness of diffraction analysis techniques such as the direct method and Patterson method in HREM is discussed. Image deconvolution and resolution enhancement for weak-phase objects by use of the direct method are illustrated. In addition, preliminary results of image restoration for thick crystals are given.

  3. High resolution CT in children with cystic fibrosis

    International Nuclear Information System (INIS)

    Stiglbauer, R.; Schurawitzki, H.; Eichler, I.; Goetz, M.

    1992-01-01

    High resolution CT (HRCT) was performed in 24 children (median age 57.9 months) suffering from cystic fibrosis (CF). In 23 patients (one examination unacceptable because of motion artifacts) the most frequent finding was bronchial wall thickening, shown in 21 patients (91%), followed by bronchiectasis in 15 patients (65%). Less frequent findings were mucus plugging and patchy consolidations, which could be demonstrated in 11 patients each (48%). Findings were classified using a CT scoring system and including only irreversible pulmonary changes; a statistically significant correlation with lung function tests could be established. HRCT to date seems to be the most valuable method to determine extent and severity of lung involvement in children with CF and should therefore be routinely used for the staging of this disease. (orig.)

  4. Chromatic Modulator for High Resolution CCD or APS Devices

    Science.gov (United States)

    Hartley, Frank T. (Inventor); Hull, Anthony B. (Inventor)

    2003-01-01

    A system for providing high-resolution color separation in electronic imaging. Comb drives controllably oscillate a red-green-blue (RGB) color strip filter system (or otherwise) over an electronic imaging system such as a charge-coupled device (CCD) or active pixel sensor (APS). The color filter is modulated over the imaging array at a rate three or more times the frame rate of the imaging array. In so doing, the underlying active imaging elements are then able to detect separate color-separated images, which are then combined to provide a color-accurate frame which is then recorded as the representation of the recorded image. High pixel resolution is maintained. Registration is obtained between the color strip filter and the underlying imaging array through the use of electrostatic comb drives in conjunction with a spring suspension system.

  5. Porous silicon phantoms for high-resolution scintillation imaging

    Energy Technology Data Exchange (ETDEWEB)

    Di Francia, G. [Portici Research Centre, ENEA, Via Vecchio Macello, 80055 Portici, Naples (Italy); Scafe, R. [Casaccia Research Centre, ENEA, 00060 S.Maria di Galeria, Rome (Italy)]. E-mail: scafe@casaccia.enea.it; De Vincentis, G. [Department of Radiological Sciences, University of Rome ' La Sapienza' , V.le Regina Elena, 324, 00161 Rome (Italy); La Ferrara, V. [Portici Research Centre, ENEA, Via Vecchio Macello, 80055 Portici, Naples (Italy); Iurlaro, G. [Casaccia Research Centre, ENEA, 00060 S.Maria di Galeria, Rome (Italy); Nasti, I. [Portici Research Centre, ENEA, Via Vecchio Macello, 80055 Portici, Naples (Italy); Montani, L. [Casaccia Research Centre, ENEA, 00060 S.Maria di Galeria, Rome (Italy); Pellegrini, R. [Department of Experimental Medicine, University of Rome ' La Sapienza' , V.le Regina Elena, 324, 00161 Rome (Italy); Betti, M. [Department of Experimental Medicine, University of Rome ' La Sapienza' , V.le Regina Elena, 324, 00161 Rome (Italy); Martucciello, N. [Portici Research Centre, ENEA, Via Vecchio Macello, 80055 Portici, Naples (Italy); Pani, R. [Department of Experimental Medicine, University of Rome ' La Sapienza' , V.le Regina Elena, 324, 00161 Rome (Italy)

    2006-12-20

    High resolution radionuclide imaging requires phantoms with precise geometries and known activities using either Anger cameras equipped with pinhole collimators or dedicated small animal devices. Porous silicon samples, having areas of different shape and size, can be made and loaded with a radioactive material, obtaining: (a) precise radio-emitting figures corresponding to the porous areas geometry (b) a radioactivity of each figure depending on the pore's specifications, and (c) the same emission energy to be used in true exams. To this aim a sample with porous circular areas has been made and loaded with a {sup 99m}TcO{sub 4} {sup -} solution. Imaging has been obtained using both general purpose and pinhole collimators. This first sample shows some defects that are analyzed and discussed.

  6. High-resolution 3-T MR neurography of peroneal neuropathy

    International Nuclear Information System (INIS)

    Chhabra, Avneesh; Faridian-Aragh, Neda; Chalian, Majid; Soldatos, Theodoros; Thawait, Shrey K.; Williams, Eric H.; Andreisek, Gustav

    2012-01-01

    The common peroneal nerve (CPN), a major terminal branch of the sciatic nerve, can be subject to a variety of pathologies, which may affect the nerve at any level from the lumbar plexus to its distal branches. Although the diagnosis of peripheral neuropathy is traditionally based on a patient's clinical findings and electrodiagnostic tests, magnetic resonance neurography (MRN) is gaining an increasing role in the definition of the type, site, and extent of peripheral nerve disorders. Current high-field MR scanners enable high-resolution and excellent soft-tissue contrast imaging of peripheral nerves. In the lower extremities, MR neurography has been employed in the demonstration of the anatomy and pathology of the CPN, as well as in the detection of associated secondary muscle denervation changes. This article reviews the normal appearance of the CPN as well as typical pathologies and abnormal findings at 3.0-T MR neurography of the lower extremity. (orig.)

  7. Meniscus tears of the knee: Postarthrogram high resolution CT

    International Nuclear Information System (INIS)

    Kim, Jae Hyoung; Do, Young Soo; You, Jin Jong; Gong, Jae Chul; Kim, Hyung Jin; Chung, Sung Hoon

    1990-01-01

    Thirty-eight knees with clinically suspected meniscal tears were examined with high resolution computed tomography(HRCT) immediately following double contrast arthrography. All subsequently underwent arthroscopy. The findings of postarthrogram HRCT and arthroscopy were compared to evaluated the usefulness of postarthrogram HRCT in diagnosis of the meniscal tears. The sensitivity, specificity and accuracy of HRCT were 96.2%. 83.3% and 92.1% respectively. The anatomic details of the meniscal tears were clearly visible on the HRCT scans. Sagittal and coronal reformation views well visualized the horizontal tears and the relationship of torn meniscal fragments, and well differential the peripheral tears from the synovial recess. Our result indicate that postarthrogram HRCT not only is a sensitive and effective method for the detection and characterization of the meniscal tears, but also provides arthroscopists with the appropriate surgical plans

  8. Quantitative high-resolution genomic analysis of single cancer cells.

    Science.gov (United States)

    Hannemann, Juliane; Meyer-Staeckling, Sönke; Kemming, Dirk; Alpers, Iris; Joosse, Simon A; Pospisil, Heike; Kurtz, Stefan; Görndt, Jennifer; Püschel, Klaus; Riethdorf, Sabine; Pantel, Klaus; Brandt, Burkhard

    2011-01-01

    During cancer progression, specific genomic aberrations arise that can determine the scope of the disease and can be used as predictive or prognostic markers. The detection of specific gene amplifications or deletions in single blood-borne or disseminated tumour cells that may give rise to the development of metastases is of great clinical interest but technically challenging. In this study, we present a method for quantitative high-resolution genomic analysis of single cells. Cells were isolated under permanent microscopic control followed by high-fidelity whole genome amplification and subsequent analyses by fine tiling array-CGH and qPCR. The assay was applied to single breast cancer cells to analyze the chromosomal region centred by the therapeutical relevant EGFR gene. This method allows precise quantitative analysis of copy number variations in single cell diagnostics.

  9. Quantitative high-resolution genomic analysis of single cancer cells.

    Directory of Open Access Journals (Sweden)

    Juliane Hannemann

    Full Text Available During cancer progression, specific genomic aberrations arise that can determine the scope of the disease and can be used as predictive or prognostic markers. The detection of specific gene amplifications or deletions in single blood-borne or disseminated tumour cells that may give rise to the development of metastases is of great clinical interest but technically challenging. In this study, we present a method for quantitative high-resolution genomic analysis of single cells. Cells were isolated under permanent microscopic control followed by high-fidelity whole genome amplification and subsequent analyses by fine tiling array-CGH and qPCR. The assay was applied to single breast cancer cells to analyze the chromosomal region centred by the therapeutical relevant EGFR gene. This method allows precise quantitative analysis of copy number variations in single cell diagnostics.

  10. High-resolution spectrum of the Galactic center

    Science.gov (United States)

    Mahoney, W. A.; Ling, J. C.; Wheaton, W. A.

    1993-01-01

    Recent observations of the Galactic center region indicate the presence of a narrow gamma-ray line feature at 170 keV, and theoretical speculations suggest it may result from Compton backscattering of the 511 keV annihilation radiation. The high-resolution gamma-ray spectrometer on HEAO 3 observed the Galactic center in the fall of 1979 and in the spring of 1980. In view of the recent developments, the HEAO data were re-examined to search for this new feature and to look for possible correlations with the 511 keV line emisison. No evidence for such Compton backscattered radiation was found and the derived upper limits for emission in a line feature near 170 keV were well below previously reported fluxes, indicating possible time variability.

  11. The inelastic contribution to high resolution images of defects

    International Nuclear Information System (INIS)

    Krivanek, O.L.; Ahn, C.C.; Wood, G.J.

    1990-01-01

    The importance of the contribution due to inelastically scattered electrons to unfiltered HREM images is examined, with emphasis on imaging of defects in semiconductors. Whenever the low energy loss spectrum contains sharp peaks, the contribution is not featureless. At specimen thickness of a few tens of nm, it may change the image appearance in a major way. The strongest effect occurs in high resolution, medium voltage (200 to 500 kV) electron microscope images of defects at focus values minimizing the contrast of the elastic image in low Z materials such as Al and Si. In higher Z materials or those with no sharp 'plasmons', the contribution is small. 23 refs., 8 figs

  12. High-Resolution Reciprocal Space Mapping for Characterizing Deformation Structures

    DEFF Research Database (Denmark)

    Pantleon, Wolfgang; Wejdemann, Christian; Jakobsen, Bo

    2014-01-01

    With high-angular resolution three-dimensional X-ray diffraction (3DXRD), quantitative information is gained about dislocation structures in individual grains in the bulk of a macroscopic specimen by acquiring reciprocal space maps. In high-resolution 3D reciprocal space maps of tensile......-deformed copper, individual, almost dislocation-free subgrains are identified from high-intensity peaks and distinguished by their unique combination of orientation and elastic strain; dislocation walls manifest themselves as a smooth cloud of lower intensity. The elastic strain shows only minor variations within...... dynamics is followed in situ during varying loading conditions by reciprocal space mapping: during uninterrupted tensile deformation, formation of subgrains is observed concurrently with broadening of Bragg reflections shortly after the onset of plastic deformation. When the traction is terminated, stress...

  13. Paracoccidioidomycosis: High-resolution computed tomography-pathologic correlation

    International Nuclear Information System (INIS)

    Marchiori, Edson; Valiante, Paulo Marcos; Mano, Claudia Mauro; Zanetti, Glaucia; Escuissato, Dante L.; Souza, Arthur Soares; Capone, Domenico

    2011-01-01

    Objective: The purpose of this study was to describe the high-resolution computed tomography (HRCT) features of pulmonary paracoccidioidomycosis and to correlate them with pathologic findings. Methods: The study included 23 adult patients with pulmonary paracoccidioidomycosis. All patients had undergone HRCT, and the images were retrospectively analyzed by two chest radiologists, who reached decisions by consensus. An experienced lung pathologist reviewed all pathological specimens. The HRCT findings were correlated with histopathologic data. Results: The predominant HRCT findings included areas of ground-glass opacities, nodules, interlobular septal thickening, airspace consolidation, cavitation, and fibrosis. The main pathological features consisted of alveolar and interlobular septal inflammatory infiltration, granulomas, alveolar exudate, cavitation secondary to necrosis, and fibrosis. Conclusion: Paracoccidioidomycosis can present different tomography patterns, which can involve both the interstitium and the airspace. These abnormalities can be pathologically correlated with inflammatory infiltration, granulomatous reaction, and fibrosis.

  14. High resolution imaging of tunnels by magnetic resonance neurography

    Energy Technology Data Exchange (ETDEWEB)

    Subhawong, Ty K.; Thawait, Shrey K.; Machado, Antonio J.; Carrino, John A.; Chhabra, Avneesh [Johns Hopkins Hospital, The Russell H. Morgan Department of Radiology and Radiological Science, Baltimore, MD (United States); Wang, Kenneth C. [Baltimore VA Medical Center, Department of Radiology, Baltimore, MD (United States); Williams, Eric H. [Dellon Institute for Peripheral Nerve Surgery, Towson, MD (United States); Hashemi, Shahreyar Shar [Johns Hopkins Hospital, Division of Plastic and Reconstructive Surgery, Baltimore, MD (United States)

    2012-01-15

    Peripheral nerves often traverse confined fibro-osseous and fibro-muscular tunnels in the extremities, where they are particularly vulnerable to entrapment and compressive neuropathy. This gives rise to various tunnel syndromes, characterized by distinct patterns of muscular weakness and sensory deficits. This article focuses on several upper and lower extremity tunnels, in which direct visualization of the normal and abnormal nerve in question is possible with high resolution 3T MR neurography (MRN). MRN can also serve as a useful adjunct to clinical and electrophysiologic exams by discriminating adhesive lesions (perineural scar) from compressive lesions (such as tumor, ganglion, hypertrophic callous, or anomalous muscles) responsible for symptoms, thereby guiding appropriate treatment. (orig.)

  15. High-resolution 3-T MR neurography of peroneal neuropathy

    Energy Technology Data Exchange (ETDEWEB)

    Chhabra, Avneesh; Faridian-Aragh, Neda; Chalian, Majid; Soldatos, Theodoros; Thawait, Shrey K. [Johns Hopkins Hospital, The Russell H. Morgan Department of Radiology and Radiological Science, Baltimore, MD (United States); Williams, Eric H. [Johns Hopkins Hospital, Department of Plastic Surgery, Baltimore, MD (United States); Dellon Institute for Peripheral Nerve Surgery, Baltimore, MD (United States); Andreisek, Gustav [University Hospital Zurich, Institute for Diagnostic Radiology, Department of Medical Radiology, Zurich (Switzerland)

    2012-03-15

    The common peroneal nerve (CPN), a major terminal branch of the sciatic nerve, can be subject to a variety of pathologies, which may affect the nerve at any level from the lumbar plexus to its distal branches. Although the diagnosis of peripheral neuropathy is traditionally based on a patient's clinical findings and electrodiagnostic tests, magnetic resonance neurography (MRN) is gaining an increasing role in the definition of the type, site, and extent of peripheral nerve disorders. Current high-field MR scanners enable high-resolution and excellent soft-tissue contrast imaging of peripheral nerves. In the lower extremities, MR neurography has been employed in the demonstration of the anatomy and pathology of the CPN, as well as in the detection of associated secondary muscle denervation changes. This article reviews the normal appearance of the CPN as well as typical pathologies and abnormal findings at 3.0-T MR neurography of the lower extremity. (orig.)

  16. High resolution imaging of tunnels by magnetic resonance neurography

    International Nuclear Information System (INIS)

    Subhawong, Ty K.; Thawait, Shrey K.; Machado, Antonio J.; Carrino, John A.; Chhabra, Avneesh; Wang, Kenneth C.; Williams, Eric H.; Hashemi, Shahreyar Shar

    2012-01-01

    Peripheral nerves often traverse confined fibro-osseous and fibro-muscular tunnels in the extremities, where they are particularly vulnerable to entrapment and compressive neuropathy. This gives rise to various tunnel syndromes, characterized by distinct patterns of muscular weakness and sensory deficits. This article focuses on several upper and lower extremity tunnels, in which direct visualization of the normal and abnormal nerve in question is possible with high resolution 3T MR neurography (MRN). MRN can also serve as a useful adjunct to clinical and electrophysiologic exams by discriminating adhesive lesions (perineural scar) from compressive lesions (such as tumor, ganglion, hypertrophic callous, or anomalous muscles) responsible for symptoms, thereby guiding appropriate treatment. (orig.)

  17. High resolution photoelectron spectroscopy of clusters of Group V elements

    International Nuclear Information System (INIS)

    Wang, Lai-sheng; Niu, B.; Lee, Y.T.; Shirley, D.A.

    1989-07-01

    High resolution HeI (580 angstrom) photoelectron spectra of As 2 , As 4 , and P 4 were obtained with a newly-built high temperature molecular beam source. Vibrational structure was resolved in the photoelectron spectra of the three cluster species. The Jahn-Teller effect is discussed for the 2 E and 2 T 2 states of P 4 + and As 4 + . As a result of the Jahn-Teller effect, the 2 E state splits into two bands, and the 2 T 2 state splits into three bands, in combination with the spin-orbit effect. It was observed that the ν 2 normal vibrational mode was involved in the vibronic interaction of the 2 E state, while both the ν 2 and ν 3 modes were active in the 2 T 2 state. 26 refs., 5 figs., 3 tabs

  18. Chandra High Resolution Imaging of NGC 1365 and NGC 4151

    Science.gov (United States)

    Wang, Junfeng; Fabbiano, G.; Elvis, M.; Risaliti, G.; Karovska, M.; Zezas, A.; Mazzarella, J. M.; Lord, S.; Howell, J. H.; Mundell, C. G.

    2010-07-01

    We present Chandra high resolution imaging of the circumnuclear regions of two nearby active galaxies, namely the starburst/AGN composite Seyfert 1.8 NGC 1365 and the archetypal Seyfert 1 NGC 4151. In NGC 1365, the X-ray morphology shows a biconical soft X-ray-emission region extending ~5 kpc in projection from the nucleus, coincident with the optical high-excitation outflows. Chandra HRC imaging of the NGC 4151 nucleus resolves X-ray emission from the 4 arcsec radio jet and the narrow line region (NLR) clouds. Our results demonstrate the unique power of spatially resolved spectroscopy with Chandra, and support previous claims that frequent jet-ISM interaction may explain why jets in Seyfert galaxies appear small, slow, and thermally dominated.

  19. High resolution ultrastructure imaging of fractures in human dental tissues

    Directory of Open Access Journals (Sweden)

    Tan Sui

    2014-01-01

    Full Text Available Human dental hard tissues are dentine, cementum, and enamel. These are hydrated mineralised composite tissues with a hierarchical structure and versatile thermo-mechanical properties. The hierarchical structure of dentine and enamel was imaged by transmission electron microscopy (TEM of samples prepared by focused ion beam (FIB milling. High resolution TEM was carried out in the vicinity of a crack tip in dentine. An intricate “random weave” pattern of hydroxyapatile crystallites was observed and this provided a possible explanation for toughening of the mineralized dentine tissue at the nano-scale. The results reported here provide the basis for improved understanding of the relationship between the multi-scale nature and the mechanical properties of hierarchically structured biomaterials, and will also be useful for the development of better prosthetic and dental restorative materials.

  20. A high resolution gridded ionization chamber for nuclear spectroscopy

    International Nuclear Information System (INIS)

    Vitale, E.R.

    1988-01-01

    This paper describes some techniques used in the design of high resolution gridded ionisation chambers for measurements of absolute activity of radionuclides. Details of the geometry of the system and its electrodes are presented; their shape and the spacing between the grid wire was studied with the help of an electrolytic tank. The experimental spectra obtained with an Am 241 source using Ar + 10% methane as a flow gas show a total resolution of 39,07 KeV in very good agreement with the best results available in the literature. An application of the methods developed was used in the design ans construction of a proportional counter provided with three sequential grids disposed in such a way that the pulses from the first stage had their amplitude multiplied by the two further stages. Multiplication factors of the order of 10 sup(3) were obtained but higher values are expected. (author)

  1. Evaluation of Esophageal Motor Function With High-resolution Manometry

    Science.gov (United States)

    2013-01-01

    For several decades esophageal manometry has been the test of choice to evaluate disorders of esophageal motor function. The recent introduction of high-resolution manometry for the study of esophageal motor function simplified performance of esophageal manometry, and revealed previously unidentified patterns of normal and abnormal esophageal motor function. Presentation of pressure data as color contour plots or esophageal pressure topography led to the development of new tools for analyzing and classifying esophageal motor patterns. The current standard and still developing approach to do this is the Chicago classification. While this methodical approach is improving our diagnosis of esophageal motor disorders, it currently does not address all motor abnormalities. We will explore the Chicago classification and disorders that it does not address. PMID:23875094

  2. High resolution imaging of surface patterns of single bacterial cells

    International Nuclear Information System (INIS)

    Greif, Dominik; Wesner, Daniel; Regtmeier, Jan; Anselmetti, Dario

    2010-01-01

    We systematically studied the origin of surface patterns observed on single Sinorhizobium meliloti bacterial cells by comparing the complementary techniques atomic force microscopy (AFM) and scanning electron microscopy (SEM). Conditions ranged from living bacteria in liquid to fixed bacteria in high vacuum. Stepwise, we applied different sample modifications (fixation, drying, metal coating, etc.) and characterized the observed surface patterns. A detailed analysis revealed that the surface structure with wrinkled protrusions in SEM images were not generated de novo but most likely evolved from similar and naturally present structures on the surface of living bacteria. The influence of osmotic stress to the surface structure of living cells was evaluated and also the contribution of exopolysaccharide and lipopolysaccharide (LPS) by imaging two mutant strains of the bacterium under native conditions. AFM images of living bacteria in culture medium exhibited surface structures of the size of single proteins emphasizing the usefulness of AFM for high resolution cell imaging.

  3. Bayesian Peptide Peak Detection for High Resolution TOF Mass Spectrometry.

    Science.gov (United States)

    Zhang, Jianqiu; Zhou, Xiaobo; Wang, Honghui; Suffredini, Anthony; Zhang, Lin; Huang, Yufei; Wong, Stephen

    2010-11-01

    In this paper, we address the issue of peptide ion peak detection for high resolution time-of-flight (TOF) mass spectrometry (MS) data. A novel Bayesian peptide ion peak detection method is proposed for TOF data with resolution of 10 000-15 000 full width at half-maximum (FWHW). MS spectra exhibit distinct characteristics at this resolution, which are captured in a novel parametric model. Based on the proposed parametric model, a Bayesian peak detection algorithm based on Markov chain Monte Carlo (MCMC) sampling is developed. The proposed algorithm is tested on both simulated and real datasets. The results show a significant improvement in detection performance over a commonly employed method. The results also agree with expert's visual inspection. Moreover, better detection consistency is achieved across MS datasets from patients with identical pathological condition.

  4. High-resolution imaging of solar system objects

    International Nuclear Information System (INIS)

    Goldberg, B.A.

    1988-01-01

    The strategy of this investigation has been to develop new high-resolution imaging capabilities and to apply them to extended observing programs. These programs have included Io's neutral sodium cloud and comets. The Io observing program was carried out at Table Mountain Observatory (1976 to 1981), providing a framework interpreting Voyager measurements of the Io torus, and serving as an important reference for studying asymmetries and time variabilities in the Jovian magnetosphere. Comet observations made with the 3.6 m Canada-France-Hawaii Telescope and 1.6 m AMOS telescope (1984 to 1987) provide basis for studying early coma development in Halley, the kinematics of its nucleus, and the internal and external structure of the nucleus. Images of GZ from the ICE encounter period form the basis for unique comparisons with in situ magnetic field and dust impact measurements to determine the ion tail and dust coma structure, respectively

  5. Polystyrene negative resist for high-resolution electron beam lithography

    Directory of Open Access Journals (Sweden)

    Ma Siqi

    2011-01-01

    Full Text Available Abstract We studied the exposure behavior of low molecular weight polystyrene as a negative tone electron beam lithography (EBL resist, with the goal of finding the ultimate achievable resolution. It demonstrated fairly well-defined patterning of a 20-nm period line array and a 15-nm period dot array, which are the densest patterns ever achieved using organic EBL resists. Such dense patterns can be achieved both at 20 and 5 keV beam energies using different developers. In addition to its ultra-high resolution capability, polystyrene is a simple and low-cost resist with easy process control and practically unlimited shelf life. It is also considerably more resistant to dry etching than PMMA. With a low sensitivity, it would find applications where negative resist is desired and throughput is not a major concern.

  6. Performance of a high resolution cavity beam position monitor system

    Science.gov (United States)

    Walston, Sean; Boogert, Stewart; Chung, Carl; Fitsos, Pete; Frisch, Joe; Gronberg, Jeff; Hayano, Hitoshi; Honda, Yosuke; Kolomensky, Yury; Lyapin, Alexey; Malton, Stephen; May, Justin; McCormick, Douglas; Meller, Robert; Miller, David; Orimoto, Toyoko; Ross, Marc; Slater, Mark; Smith, Steve; Smith, Tonee; Terunuma, Nobuhiro; Thomson, Mark; Urakawa, Junji; Vogel, Vladimir; Ward, David; White, Glen

    2007-07-01

    It has been estimated that an RF cavity Beam Position Monitor (BPM) could provide a position measurement resolution of less than 1 nm. We have developed a high resolution cavity BPM and associated electronics. A triplet comprised of these BPMs was installed in the extraction line of the Accelerator Test Facility (ATF) at the High Energy Accelerator Research Organization (KEK) for testing with its ultra-low emittance beam. The three BPMs were each rigidly mounted inside an alignment frame on six variable-length struts which could be used to move the BPMs in position and angle. We have developed novel methods for extracting the position and tilt information from the BPM signals including a robust calibration algorithm which is immune to beam jitter. To date, we have demonstrated a position resolution of 15.6 nm and a tilt resolution of 2.1 μrad over a dynamic range of approximately ±20 μm.

  7. High resolution 3D imaging of synchrotron generated microbeams

    Energy Technology Data Exchange (ETDEWEB)

    Gagliardi, Frank M., E-mail: frank.gagliardi@wbrc.org.au [Alfred Health Radiation Oncology, The Alfred, Melbourne, Victoria 3004, Australia and School of Medical Sciences, RMIT University, Bundoora, Victoria 3083 (Australia); Cornelius, Iwan [Imaging and Medical Beamline, Australian Synchrotron, Clayton, Victoria 3168, Australia and Centre for Medical Radiation Physics, University of Wollongong, Wollongong, New South Wales 2500 (Australia); Blencowe, Anton [Division of Health Sciences, School of Pharmacy and Medical Sciences, The University of South Australia, Adelaide, South Australia 5000, Australia and Division of Information Technology, Engineering and the Environment, Mawson Institute, University of South Australia, Mawson Lakes, South Australia 5095 (Australia); Franich, Rick D. [School of Applied Sciences and Health Innovations Research Institute, RMIT University, Melbourne, Victoria 3000 (Australia); Geso, Moshi [School of Medical Sciences, RMIT University, Bundoora, Victoria 3083 (Australia)

    2015-12-15

    Purpose: Microbeam radiation therapy (MRT) techniques are under investigation at synchrotrons worldwide. Favourable outcomes from animal and cell culture studies have proven the efficacy of MRT. The aim of MRT researchers currently is to progress to human clinical trials in the near future. The purpose of this study was to demonstrate the high resolution and 3D imaging of synchrotron generated microbeams in PRESAGE® dosimeters using laser fluorescence confocal microscopy. Methods: Water equivalent PRESAGE® dosimeters were fabricated and irradiated with microbeams on the Imaging and Medical Beamline at the Australian Synchrotron. Microbeam arrays comprised of microbeams 25–50 μm wide with 200 or 400 μm peak-to-peak spacing were delivered as single, cross-fire, multidirectional, and interspersed arrays. Imaging of the dosimeters was performed using a NIKON A1 laser fluorescence confocal microscope. Results: The spatial fractionation of the MRT beams was clearly visible in 2D and up to 9 mm in depth. Individual microbeams were easily resolved with the full width at half maximum of microbeams measured on images with resolutions of as low as 0.09 μm/pixel. Profiles obtained demonstrated the change of the peak-to-valley dose ratio for interspersed MRT microbeam arrays and subtle variations in the sample positioning by the sample stage goniometer were measured. Conclusions: Laser fluorescence confocal microscopy of MRT irradiated PRESAGE® dosimeters has been validated in this study as a high resolution imaging tool for the independent spatial and geometrical verification of MRT beam delivery.

  8. Evaluation of a High-Resolution Regional Reanalysis for Europe

    Science.gov (United States)

    Ohlwein, C.; Wahl, S.; Keller, J. D.; Bollmeyer, C.

    2014-12-01

    Reanalyses gain more and more importance as a source of meteorological information for many purposes and applications. Several global reanalyses projects (e.g., ERA, MERRA, CSFR, JMA9) produce and verify these data sets to provide time series as long as possible combined with a high data quality. Due to a spatial resolution down to 50-70km and 3-hourly temporal output, they are not suitable for small scale problems (e.g., regional climate assessment, meso-scale NWP verification, input for subsequent models such as river runoff simulations). The implementation of regional reanalyses based on a limited area model along with a data assimilation scheme is able to generate reanalysis data sets with high spatio-temporal resolution. Within the Hans-Ertel-Centre for Weather Research (HErZ), the climate monitoring branch concentrates efforts on the assessment and analysis of regional climate in Germany and Europe. In joint cooperation with DWD (German Meteorological Service), a high-resolution reanalysis system based on the COSMO model has been developed. The regional reanalysis for Europe matches the domain of the CORDEX EURO-11 specifications, albeit at a higher spatial resolution, i.e., 0.055° (6km) instead of 0.11° (12km) and comprises the assimilation of observational data using the existing nudging scheme of COSMO complemented by a special soil moisture analysis with boundary conditions provided by ERA-Interim data. The reanalysis data set covers 6 years (2007-2012) and is currently extended to 16 years. Extensive evaluation of the reanalysis is performed using independent observations with special emphasis on precipitation and high-impact weather situations indicating a better representation of small scale variability. Further, the evaluation shows an added value of the regional reanalysis with respect to the forcing ERA Interim reanalysis and compared to a pure high-resolution dynamical downscaling approach without data assimilation.

  9. The high-resolution regional reanalysis COSMO-REA6

    Science.gov (United States)

    Ohlwein, C.

    2016-12-01

    Reanalyses gain more and more importance as a source of meteorological information for many purposes and applications. Several global reanalyses projects (e.g., ERA, MERRA, CSFR, JMA9) produce and verify these data sets to provide time series as long as possible combined with a high data quality. Due to a spatial resolution down to 50-70km and 3-hourly temporal output, they are not suitable for small scale problems (e.g., regional climate assessment, meso-scale NWP verification, input for subsequent models such as river runoff simulations). The implementation of regional reanalyses based on a limited area model along with a data assimilation scheme is able to generate reanalysis data sets with high spatio-temporal resolution. Within the Hans-Ertel-Centre for Weather Research (HErZ), the climate monitoring branch concentrates efforts on the assessment and analysis of regional climate in Germany and Europe. In joint cooperation with DWD (German Meteorological Service), a high-resolution reanalysis system based on the COSMO model has been developed. The regional reanalysis for Europe matches the domain of the CORDEX EURO-11 specifications, albeit at a higher spatial resolution, i.e., 0.055° (6km) instead of 0.11° (12km) and comprises the assimilation of observational data using the existing nudging scheme of COSMO complemented by a special soil moisture analysis with boundary conditions provided by ERA-Interim data. The reanalysis data set covers the past 20 years. Extensive evaluation of the reanalysis is performed using independent observations with special emphasis on precipitation and high-impact weather situations indicating a better representation of small scale variability. Further, the evaluation shows an added value of the regional reanalysis with respect to the forcing ERA Interim reanalysis and compared to a pure high-resolution dynamical downscaling approach without data assimilation.

  10. A High-resolution Reanalysis for the European CORDEX Region

    Science.gov (United States)

    Bentzien, Sabrina; Bollmeyer, Christoph; Crewell, Susanne; Friederichs, Petra; Hense, Andreas; Keller, Jan; Keune, Jessica; Kneifel, Stefan; Ohlwein, Christian; Pscheidt, Ieda; Redl, Stephanie; Steinke, Sandra

    2014-05-01

    A High-resolution Reanalysis for the European CORDEX Region Within the Hans-Ertel-Centre for Weather Research (HErZ), the climate monitoring branch concentrates efforts on the assessment and analysis of regional climate in Germany and Europe. In joint cooperation with DWD (German Meteorological Service), a high-resolution reanalysis system based on the COSMO model has been developed. Reanalyses gain more and more importance as a source of meteorological information for many purposes and applications. Several global reanalyses projects (e.g., ERA, MERRA, CSFR, JMA9) produce and verify these data sets to provide time series as long as possible combined with a high data quality. Due to a spatial resolution down to 50-70km and 3-hourly temporal output, they are not suitable for small scale problems (e.g., regional climate assessment, meso-scale NWP verification, input for subsequent models such as river runoff simulations). The implementation of regional reanalyses based on a limited area model along with a data assimilation scheme is able to generate reanalysis data sets with high spatio-temporal resolution. The work presented here focuses on the regional reanalysis for Europe with a domain matching the CORDEX-EURO-11 specifications, albeit at a higher spatial resolution, i.e., 0.055° (6km) instead of 0.11° (12km). The COSMO reanalysis system comprises the assimilation of observational data using the existing nudging scheme of COSMO and is complemented by a special soil moisture analysis and boundary conditions given by ERA-interim data. The reanalysis data set currently covers 6 years (2007-2012). The evaluation of the reanalyses is done using independent observations with special emphasis on precipitation and high-impact weather situations. The development and evaluation of the COSMO-based reanalysis for the CORDEX-Euro domain can be seen as a preparation for joint European activities on the development of an ensemble system of regional reanalyses for Europe.

  11. A high-resolution regional reanalysis for Europe

    Science.gov (United States)

    Ohlwein, C.

    2015-12-01

    Reanalyses gain more and more importance as a source of meteorological information for many purposes and applications. Several global reanalyses projects (e.g., ERA, MERRA, CSFR, JMA9) produce and verify these data sets to provide time series as long as possible combined with a high data quality. Due to a spatial resolution down to 50-70km and 3-hourly temporal output, they are not suitable for small scale problems (e.g., regional climate assessment, meso-scale NWP verification, input for subsequent models such as river runoff simulations). The implementation of regional reanalyses based on a limited area model along with a data assimilation scheme is able to generate reanalysis data sets with high spatio-temporal resolution. Within the Hans-Ertel-Centre for Weather Research (HErZ), the climate monitoring branch concentrates efforts on the assessment and analysis of regional climate in Germany and Europe. In joint cooperation with DWD (German Meteorological Service), a high-resolution reanalysis system based on the COSMO model has been developed. The regional reanalysis for Europe matches the domain of the CORDEX EURO-11 specifications, albeit at a higher spatial resolution, i.e., 0.055° (6km) instead of 0.11° (12km) and comprises the assimilation of observational data using the existing nudging scheme of COSMO complemented by a special soil moisture analysis with boundary conditions provided by ERA-Interim data. The reanalysis data set covers the past 20 years. Extensive evaluation of the reanalysis is performed using independent observations with special emphasis on precipitation and high-impact weather situations indicating a better representation of small scale variability. Further, the evaluation shows an added value of the regional reanalysis with respect to the forcing ERA Interim reanalysis and compared to a pure high-resolution dynamical downscaling approach without data assimilation.

  12. High-resolution RCMs as pioneers for future GCMs

    Science.gov (United States)

    Schar, C.; Ban, N.; Arteaga, A.; Charpilloz, C.; Di Girolamo, S.; Fuhrer, O.; Hoefler, T.; Leutwyler, D.; Lüthi, D.; Piaget, N.; Ruedisuehli, S.; Schlemmer, L.; Schulthess, T. C.; Wernli, H.

    2017-12-01

    Currently large efforts are underway to refine the horizontal resolution of global and regional climate models to O(1 km), with the intent to represent convective clouds explicitly rather than using semi-empirical parameterizations. This refinement will move the governing equations closer to first principles and is expected to reduce the uncertainties of climate models. High resolution is particularly attractive in order to better represent critical cloud feedback processes (e.g. related to global climate sensitivity and extratropical summer convection) and extreme events (such as heavy precipitation events, floods, and hurricanes). The presentation will be illustrated using decade-long simulations at 2 km horizontal grid spacing, some of these covering the European continent on a computational mesh with 1536x1536x60 grid points. To accomplish such simulations, use is made of emerging heterogeneous supercomputing architectures, using a version of the COSMO limited-area weather and climate model that is able to run entirely on GPUs. Results show that kilometer-scale resolution dramatically improves the simulation of precipitation in terms of the diurnal cycle and short-term extremes. The modeling framework is used to address changes of precipitation scaling with climate change. It is argued that already today, modern supercomputers would in principle enable global atmospheric convection-resolving climate simulations, provided appropriately refactored codes were available, and provided solutions were found to cope with the rapidly growing output volume. A discussion will be provided of key challenges affecting the design of future high-resolution climate models. It is suggested that km-scale RCMs should be exploited to pioneer this terrain, at a time when GCMs are not yet available at such resolutions. Areas of interest include the development of new parameterization schemes adequate for km-scale resolution, the exploration of new validation methodologies and data

  13. A high-resolution multimode digital microscope system.

    Science.gov (United States)

    Salmon, Edward D; Shaw, Sidney L; Waters, Jennifer C; Waterman-Storer, Clare M; Maddox, Paul S; Yeh, Elaine; Bloom, Kerry

    2013-01-01

    This chapter describes the development of a high-resolution, multimode digital imaging system based on a wide-field epifluorescent and transmitted light microscope, and a cooled charge-coupled device (CCD) camera. The three main parts of this imaging system are Nikon FXA microscope, Hamamatsu C4880 cooled CCD camera, and MetaMorph digital imaging system. This chapter presents various design criteria for the instrument and describes the major features of the microscope components-the cooled CCD camera and the MetaMorph digital imaging system. The Nikon FXA upright microscope can produce high resolution images for both epifluorescent and transmitted light illumination without switching the objective or moving the specimen. The functional aspects of the microscope set-up can be considered in terms of the imaging optics, the epi-illumination optics, the transillumination optics, the focus control, and the vibration isolation table. This instrument is somewhat specialized for microtubule and mitosis studies, and it is also applicable to a variety of problems in cellular imaging, including tracking proteins fused to the green fluorescent protein in live cells. The instrument is also valuable for correlating the assembly dynamics of individual cytoplasmic microtubules (labeled by conjugating X-rhodamine to tubulin) with the dynamics of membranes of the endoplasmic reticulum (labeled with DiOC6) and the dynamics of the cell cortex (by differential interference contrast) in migrating vertebrate epithelial cells. This imaging system also plays an important role in the analysis of mitotic mutants in the powerful yeast genetic system Saccharomyces cerevisiae. Copyright © 1998 Elsevier Inc. All rights reserved.

  14. Enhancing GIS Capabilities for High Resolution Earth Science Grids

    Science.gov (United States)

    Koziol, B. W.; Oehmke, R.; Li, P.; O'Kuinghttons, R.; Theurich, G.; DeLuca, C.

    2017-12-01

    Applications for high performance GIS will continue to increase as Earth system models pursue more realistic representations of Earth system processes. Finer spatial resolution model input and output, unstructured or irregular modeling grids, data assimilation, and regional coordinate systems present novel challenges for GIS frameworks operating in the Earth system modeling domain. This presentation provides an overview of two GIS-driven applications that combine high performance software with big geospatial datasets to produce value-added tools for the modeling and geoscientific community. First, a large-scale interpolation experiment using National Hydrography Dataset (NHD) catchments, a high resolution rectilinear CONUS grid, and the Earth System Modeling Framework's (ESMF) conservative interpolation capability will be described. ESMF is a parallel, high-performance software toolkit that provides capabilities (e.g. interpolation) for building and coupling Earth science applications. ESMF is developed primarily by the NOAA Environmental Software Infrastructure and Interoperability (NESII) group. The purpose of this experiment was to test and demonstrate the utility of high performance scientific software in traditional GIS domains. Special attention will be paid to the nuanced requirements for dealing with high resolution, unstructured grids in scientific data formats. Second, a chunked interpolation application using ESMF and OpenClimateGIS (OCGIS) will demonstrate how spatial subsetting can virtually remove computing resource ceilings for very high spatial resolution interpolation operations. OCGIS is a NESII-developed Python software package designed for the geospatial manipulation of high-dimensional scientific datasets. An overview of the data processing workflow, why a chunked approach is required, and how the application could be adapted to meet operational requirements will be discussed here. In addition, we'll provide a general overview of OCGIS

  15. COMPARATIVE ASSESSMENT OF VERY HIGH RESOLUTION SATELLITE AND AERIAL ORTHOIMAGERY

    Directory of Open Access Journals (Sweden)

    P. Agrafiotis

    2015-03-01

    Full Text Available This paper aims to assess the accuracy and radiometric quality of orthorectified high resolution satellite imagery from Pleiades-1B satellites through a comparative evaluation of their quantitative and qualitative properties. A Pleiades-B1 stereopair of high resolution images taken in 2013, two adjacent GeoEye-1 stereopairs from 2011 and aerial orthomosaic (LSO provided by NCMA S.A (Hellenic Cadastre from 2007 have been used for the comparison tests. As control dataset orthomosaic from aerial imagery provided also by NCMA S.A (0.25m GSD from 2012 was selected. The process for DSM and orthoimage production was performed using commercial digital photogrammetric workstations. The two resulting orthoimages and the aerial orthomosaic (LSO were relatively and absolutely evaluated for their quantitative and qualitative properties. Test measurements were performed using the same check points in order to establish their accuracy both as far as the single point coordinates as well as their distances are concerned. Check points were distributed according to JRC Guidelines for Best Practice and Quality Checking of Ortho Imagery and NSSDA standards while areas with different terrain relief and land cover were also included. The tests performed were based also on JRC and NSSDA accuracy standards. Finally, tests were carried out in order to assess the radiometric quality of the orthoimagery. The results are presented with a statistical analysis and they are evaluated in order to present the merits and demerits of the imaging sensors involved for orthoimage production. The results also serve for a critical approach for the usability and cost efficiency of satellite imagery for the production of Large Scale Orthophotos.

  16. High resolution micro ultrasonic machining for trimming 3D microstructures

    International Nuclear Information System (INIS)

    Viswanath, Anupam; Li, Tao; Gianchandani, Yogesh

    2014-01-01

    This paper reports on the evaluation of a high resolution micro ultrasonic machining (HR-µUSM) process suitable for post fabrication trimming of complex 3D microstructures made from fused silica. Unlike conventional USM, the HR-µUSM process aims for low machining rates, providing high resolution and high surface quality. The machining rate is reduced by keeping the micro-tool tip at a fixed distance from the workpiece and vibrating it at a small amplitude. The surface roughness is improved by an appropriate selection of abrasive particles. Fluidic modeling is performed to study interaction among the vibrating micro-tool tip, workpiece, and the slurry. Using 304 stainless steel (SS304) tool tips of 50 µm diameter, the machining performance of the HR-µUSM process is characterized on flat fused silica substrates. The depths and surface finish of machined features are evaluated as functions of slurry concentrations, separation between the micro-tool and workpiece, and machining time. Under the selected conditions, the HR-µUSM process achieves machining rates as low as 10 nm s −1  averaged over the first minute of machining of a flat virgin sample. This corresponds to a mass removal rate of ≈20 ng min −1 . The average surface roughness, S a , achieved is as low as 30 nm. Analytical and numerical modeling are used to explain the typical profile of the machined features as well as machining rates. The process is used to demonstrate trimming of hemispherical 3D shells made of fused silica. (paper)

  17. High resolution 3D imaging of synchrotron generated microbeams

    International Nuclear Information System (INIS)

    Gagliardi, Frank M.; Cornelius, Iwan; Blencowe, Anton; Franich, Rick D.; Geso, Moshi

    2015-01-01

    Purpose: Microbeam radiation therapy (MRT) techniques are under investigation at synchrotrons worldwide. Favourable outcomes from animal and cell culture studies have proven the efficacy of MRT. The aim of MRT researchers currently is to progress to human clinical trials in the near future. The purpose of this study was to demonstrate the high resolution and 3D imaging of synchrotron generated microbeams in PRESAGE® dosimeters using laser fluorescence confocal microscopy. Methods: Water equivalent PRESAGE® dosimeters were fabricated and irradiated with microbeams on the Imaging and Medical Beamline at the Australian Synchrotron. Microbeam arrays comprised of microbeams 25–50 μm wide with 200 or 400 μm peak-to-peak spacing were delivered as single, cross-fire, multidirectional, and interspersed arrays. Imaging of the dosimeters was performed using a NIKON A1 laser fluorescence confocal microscope. Results: The spatial fractionation of the MRT beams was clearly visible in 2D and up to 9 mm in depth. Individual microbeams were easily resolved with the full width at half maximum of microbeams measured on images with resolutions of as low as 0.09 μm/pixel. Profiles obtained demonstrated the change of the peak-to-valley dose ratio for interspersed MRT microbeam arrays and subtle variations in the sample positioning by the sample stage goniometer were measured. Conclusions: Laser fluorescence confocal microscopy of MRT irradiated PRESAGE® dosimeters has been validated in this study as a high resolution imaging tool for the independent spatial and geometrical verification of MRT beam delivery

  18. High resolution microphotonic needle for endoscopic imaging (Conference Presentation)

    Science.gov (United States)

    Tadayon, Mohammad Amin; Mohanty, Aseema; Roberts, Samantha P.; Barbosa, Felippe; Lipson, Michal

    2017-02-01

    GRIN (Graded index) lens have revolutionized micro endoscopy enabling deep tissue imaging with high resolution. The challenges of traditional GRIN lenses are their large size (when compared with the field of view) and their limited resolution. This is because of the relatively weak NA in standard graded index lenses. Here we introduce a novel micro-needle platform for endoscopy with much higher resolution than traditional GRIN lenses and a FOV that corresponds to the whole cross section of the needle. The platform is based on polymeric (SU-8) waveguide integrated with a microlens micro fabricated on a silicon substrate using a unique molding process. Due to the high index of refraction of the material the NA of the needle is much higher than traditional GRIN lenses. We tested the probe in a fluorescent dye solution (19.6 µM Alexa Flour 647 solution) and measured a numerical aperture of 0.25, focal length of about 175 µm and minimal spot size of about 1.6 µm. We show that the platform can image a sample with the field of view corresponding to the cross sectional area of the waveguide (80x100 µm2). The waveguide size can in principle be modified to vary size of the imaging field of view. This demonstration, combined with our previous work demonstrating our ability to implant the high NA needle in a live animal, shows that the proposed system can be used for deep tissue imaging with very high resolution and high field of view.

  19. Strengthening IAEA safeguards using high-resolution commercial satellite imagery

    International Nuclear Information System (INIS)

    Zhang Hui

    2001-01-01

    Full text: In May 1997, the IAEA Board of Governors adopted the Additional Safeguards Protocol to improve its ability to detect the undeclared production of fissile material. This new strengthened safeguards system has opened the door for the IAEA to use of all types of information, including the potential use of commercial satellite imagery. We have therefore been investigating the feasibility of strengthening IAEA safeguards using commercial satellite imagery. Based on our analysis on a number of one-meter resolution IKONOS satellite images of military nuclear production facilities at nuclear states including Russia, China, India, Pakistan and Israel, we found that the new high-resolution commercial satellite imagery would play a new and valuable role in strengthening IAEA safeguards. Since 1999, images with a resolution of one meter have been available commercially from Space Imaging's IKONOS satellite. One-meter images from other companies are expected to enter the market soon. Although still an order of magnitude less capable than military imaging satellites, the capabilities of these new high-resolution commercial satellites are good enough to detect and identify the major visible characteristics of nuclear production facilities and sites. Unlike the classified spy satellite photos limited to few countries, the commercial satellite imagery is commercially available to anyone who wants to purchase it. Therefore, the new commercial satellite open a new chance that each state, international organizations, and non-governmental groups could use the commercial images to play a more proactive role in monitoring the nuclear activities in related countries and verifying the compliance of non-proliferation agreements. This could help galvanize support for intensified efforts to slow the pace of nuclear proliferation. To produce fissile materials (plutonium and highly enriched uranium) for weapons, a country would operate dedicated plutonium-production reactors and the

  20. Are There Mutator Polymerases?

    Directory of Open Access Journals (Sweden)

    Miguel Garcia-Diaz

    2003-01-01

    Full Text Available DNA polymerases are involved in different cellular events, including genome replication and DNA repair. In the last few years, a large number of novel DNA polymerases have been discovered, and the biochemical analysis of their properties has revealed a long list of intriguing features. Some of these polymerases have a very low fidelity and have been suggested to play mutator roles in different processes, like translesion synthesis or somatic hypermutation. The current view of these processes is reviewed, and the current understanding of DNA polymerases and their role as mutator enzymes is discussed.