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Sample records for sybr green-based real

  1. Rapid and Specific Detection of Acidovorax avenae subsp. citrulli Using SYBR Green-Based Real-Time PCR Amplification of the YD-Repeat Protein Gene.

    Science.gov (United States)

    Cho, Min Seok; Park, Duck Hwan; Ahn, Tae-Young; Park, Dong Suk

    2015-09-01

    The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10(0) fg/μl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

  2. Development and evaluation of a SYBR green-based real time RT-PCR assay for detection of the emerging avian influenza A (H7N9 virus.

    Directory of Open Access Journals (Sweden)

    Zheng Zhu

    Full Text Available Most recently a novel avian-origin influenza A (H7N9 virus emerged in China and has been associated with lots of human infection and fatal cases. Genetic analysis of the viral genome revealed that this reassortant virus might be better adapted to humans than other avian influenza viruses. Molecular diagnostic methods are thus urgently needed in public health laboratories. In this study, a SYBR green-based one-step real time reverse transcription-PCR (RT-PCR was developed to detect the novel H7N9 virus. The primer pairs on the basis of the hemagglutinin and neuraminidase gene sequences of H7N9 viruses amplified subtype-specific fragments with Tm values of 80.77±0.06°C for H7 and 81.20±0.17°C for N9 respectively. The standard curves showed a dynamic linear range across 6 log units of RNA copy number (10(6 to 10(1 copies/ µl with a detection limit of 10 copies per reaction for both H7 and N9 assays by using serial ten-fold diluted in-vitro transcribed viral RNA. In addition, no cross-reactivity was observed with seasonal H1N1, H1N1 pdm09, H3N2, H5N1 and H9N2 viruses as well as other human respiratory viruses. When the assay was further evaluated in H7N9 virus infected clinical samples, positive amplification signals were obtained in all of the specimens with the accordance between H7 and N9 assays. Therefore, the established SYBR green-based real time RT-PCR assay could provide a rapid, sensitive, specific and reliable alternative approach with lower costs for high throughput screening of suspected samples from humans, animals and environments in first line public health laboratories.

  3. An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.

    Science.gov (United States)

    Encinas-García, Trinidad; Mendoza-Cano, Fernando; Enríquez-Espinoza, Tania; Luken-Vega, Leonardo; Vichido-Chávez, Rodrigo; Sánchez-Paz, Arturo

    2015-02-01

    The Penaeus stylirostris densovirus (PstDV) (also known as infectious hypodermal and hematopoietic necrosis virus, IHHNV), one of the major shrimp pathogens, has a worldwide distribution in farmed and wild shrimp populations. Outbreaks of IHHNV have been associated with substantial economic losses which are accompanied by a negative social impact. Current diagnostic PCR tests may result in false-positive results as several parts of PstDV genome may be endogenized in the nuclear genome of the shrimp P. stylirostris. A one-step qPCR SYBR-Green based quantitative real-time polymerase chain reaction (qPCR) assay to detect different isolates of the IHHNV in shrimp samples was developed. The detection limit of the assay was 81 viral copies of targeted DNA per reaction. The specificity of the assay was evaluated by melting curve analysis, which showed that the IHHNV product generated a single melt peak at 81.4±0.044°C. The assay was more sensitive than conventional PCR. The standardized PCR was shown to be highly sensible, specific, robust, and reproducible, which makes it an economical and powerful tool for both diagnostic applications and general research of IHHNV. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. A novel rapid genotyping technique for Collie eye anomaly: SYBR Green-based real-time polymerase chain reaction method applicable to blood and saliva specimens on Flinders Technology Associates filter paper.

    Science.gov (United States)

    Chang, Hye-Sook; Mizukami, Keijiro; Yabuki, Akira; Hossain, Mohammad A; Rahman, Mohammad M; Uddin, Mohammad M; Arai, Toshiro; Yamato, Osamu

    2010-09-01

    Collie eye anomaly (CEA) is a canine inherited ocular disease that shows a wide variety of manifestations and severity of clinical lesions. Recently, a CEA-associated mutation was reported, and a DNA test that uses conventional polymerase chain reaction (PCR) has now become available. The objective of the current study was to develop a novel rapid genotyping technique by using SYBR Green-based real-time PCR for future large-scale surveys as a key part in the strategy to eradicate CEA by selective breeding. First, a SYBR Green-based real-time PCR assay for genotyping of CEA was developed and evaluated by using purified DNA samples from normal, carrier, and affected Border Collies in which genotypes had previously been determined by conventional PCR. This real-time PCR assay demonstrated appropriate amplifications in all genotypes, and the results were consistent with those of conventional PCR. Second, the availability of Flinders Technology Associates filter paper (FTA card) as DNA templates for the real-time PCR assay was evaluated by using blood and saliva specimens to determine suitability for CEA screening. DNA-containing solution prepared from a disc of blood- or saliva-spotted FTA cards was available directly as templates for the real-time PCR assay when the volume of solution was 2.5% of the PCR mixture. In conclusion, SYBR Green-based real-time PCR combined with FTA cards is a rapid genotyping technique for CEA that can markedly shorten the overall time required for genotyping as well as simplify the sample preparation. Therefore, this newly developed technique suits large-scale screening in breeding populations of Collie-related breeds.

  5. 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

  6. SYBR green-based real-time reverse transcription-PCR for typing and subtyping of all hemagglutinin and neuraminidase genes of avian influenza viruses and comparison to standard serological subtyping tests

    Science.gov (United States)

    Tsukamoto, K.; Javier, P.C.; Shishido, M.; Noguchi, D.; Pearce, J.; Kang, H.-M.; Jeong, O.M.; Lee, Y.-J.; Nakanishi, K.; Ashizawa, T.

    2012-01-01

    Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10 1.5, 10 2.3, and 10 3.1 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples. Copyright ?? 2012, American Society for Microbiology. All Rights Reserved.

  7. Optimization of the Divergent method for genotyping single nucleotide variations using SYBR Green-based single-tube real-time PCR.

    Science.gov (United States)

    Gentilini, Fabio; Turba, Maria E

    2014-01-01

    A novel technique, called Divergent, for single-tube real-time PCR genotyping of point mutations without the use of fluorescently labeled probes has recently been reported. This novel PCR technique utilizes a set of four primers and a particular denaturation temperature for simultaneously amplifying two different amplicons which extend in opposite directions from the point mutation. The two amplicons can readily be detected using the melt curve analysis downstream to a closed-tube real-time PCR. In the present study, some critical aspects of the original method were specifically addressed to further implement the technique for genotyping the DNM1 c.G767T mutation responsible for exercise-induced collapse in Labrador retriever dogs. The improved Divergent assay was easily set up using a standard two-step real-time PCR protocol. The melting temperature difference between the mutated and the wild-type amplicons was approximately 5°C which could be promptly detected by all the thermal cyclers. The upgraded assay yielded accurate results with 157pg of genomic DNA per reaction. This optimized technique represents a flexible and inexpensive alternative to the minor grove binder fluorescently labeled method and to high resolution melt analysis for high-throughput, robust and cheap genotyping of single nucleotide variations. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis.

    Science.gov (United States)

    Behets, Jonas; Declerck, Priscilla; Delaedt, Yasmine; Verelst, Lieve; Ollevier, Frans

    2006-12-01

    Real-time polymerase chain reaction melting curve analysis (MCA) allows differentiation of several free-living amoebae species. Distinctive characteristics were found for Naegleria fowleri, N. lovaniensis, N. australiensis, N. gruberi, Hartmanella vermiformis, and Willaertia magna. Species specificity of the amplicons was confirmed using agarose gel electrophoresis and sequence-based approaches. Amplification efficiency ranged from 91% to 98%, indicating the quantitative potential of the assay. This MCA approach can be used for quantitative detection of free-living amoebae after cultivation but also as a culture-independent detection method.

  9. SYBR green-based detection of Leishmania infantum DNA using peripheral blood samples.

    Science.gov (United States)

    Ghasemian, Mehrdad; Gharavi, Mohammad Javad; Akhlaghi, Lame; Mohebali, Mehdi; Meamar, Ahmad Reza; Aryan, Ehsan; Oormazdi, Hormozd; Ghayour, Zahra

    2016-03-01

    Parasitological methods for the diagnosis of visceral leishmaniasis (VL) require invasive sampling procedures. The aim of this study was to detect Leishmania infantum (L. infantum) DNA by real time-PCR method in peripheral blood of symptomatic VL patient and compared its performance with nested PCR, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed VL patients diagnosed by direct agglutination test (DAT > 3200), bone marrow aspiration and presented characteristic clinical features (fever, hepatosplenomegaly, and anemia) and 40 controls (non-endemic healthy control-30, Malaria-2, Toxoplasma gondii-2, Mycobacterium tuberculosis-2, HBV-1, HCV-1, HSV-1 and CMV-1) were enrolled in this study. SYBR-green based real time-PCR and nested PCR was performed to amplify the Kinetoplast DNA minicircle gene using the DNA extracted from Buffy coat. From among 47 patients, 45 (95.7 %) were positive by both nested-PCR and real time-PCR. These results indicate that real time-PCR was not only as sensitive as a nested-PCR assay for detection of Leishmania kDNA in clinical sample, but also more rapid. The advantage of real time-PCR based methods over nested-PCR is simple to perform, more faster in which nested-PCR requires post-PCR processing and reducing contamination risk.

  10. Investigation of a Sybr-Green-Based Method to Validate DNA Sequences for DNA Computing

    National Research Council Canada - National Science Library

    Pogozelski, Wendy; Priore, Salvatore; Bernard, Matthew; Macula, Anthony

    2005-01-01

    .... Strands were tested for their potential to cross-hybridize by measuring fluorescence over varying temperatures in the presence of SYBR Green Strands were tested in pools to show that hybridization...

  11. Detection of Campylobacter jejuni in dairy farm environmental samples using SYBR Green real-time polymerase chain reaction.

    Science.gov (United States)

    Nam, H M; Srinivasan, V; Murinda, S E; Oliver, S P

    2005-01-01

    The aim of this study was to evaluate a SYBR Green based real-time PCR assay using well-characterized primers to detect Campylobacter jejuni in naturally contaminated dairy farm environmental samples. Specificity of the assay was determined with 62 C. jejuni strains and 120 non-C. jejuni strains. Peak melting temperature obtained with melting curves specific for C. jejuni was 77.5 degrees C. Standard curves were constructed using mean threshold cycle (C(T)) and various concentrations of C. jejuni ranging from 10(0) to 10(8) colony forming units (CFU)/mL, which resulted in a linear relationship between C(T) and log input DNA. Correlation coefficients of standard curves based on pure culture of C. jejuni in broth and spiked cells in lagoon water were R(2) = 0.995 (slope = 3.21) and R(2) = 0.988 (slope = 3.22), respectively, and sensitivity limits were 10(3) CFU/mL, respectively. After 24-h enrichment, total C. jejuni counts of all samples spiked with 10(0) CFU/mL reached >10(5) CFU/mL, and the detection limit was improved from >10(3) CFU/mL to PCR assay detected C. jejuni in 25 (30.4%) of 82 samples, with 17 (68%) of these samples being culture positive for C. jejuni. All samples that were positive by standard culture methods were also positive by the real-time PCR method. Mean C( T ) values of 48-h enriched cultures for 17 PCR-positive/culture-positive samples and eight PCR-positive/culture-negative samples were 21.4 +/- 3.6, and 34.6 +/- 1.5 (p 38.0. These results indicate that the SYBR Green real-time PCR assay provides a specific, reproducible, and simple method for detecting C. jejuni in dairy farm environmental samples.

  12. Identifying Haemophilus haemolyticus and Haemophilus influenzae by SYBR Green real-time PCR.

    Science.gov (United States)

    Latham, Roger; Zhang, Bowen; Tristram, Stephen

    2015-05-01

    SYBR Green real time PCR assays for protein D (hpd), fuculose kinase (fucK) and [Cu, Zn]-superoxide dismutase (sodC) were designed for use in an algorithm for the identification of Haemophilus influenzae and H. haemolyticus. When tested on 127 H. influenzae and 60 H. haemolyticus all isolates were identified correctly. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  13. A Multiplex SYBR Green Real-Time PCR Assay for the Detection of Three Colistin Resistance Genes from Cultured Bacteria, Feces, and Environment Samples

    Directory of Open Access Journals (Sweden)

    Jiyun Li

    2017-10-01

    Full Text Available The aim of the study was to develop a multiplex assay for rapid detection of mcr-1, mcr-2, and mcr-3, a group of genes of conferring resistance to colistin mediated by plasmid in Enterobacteriaceae. A SYBR Green based real-time PCR assay has been designed to detect the mcr genes, and applied to cultured bacteria, feces and soil samples. All three mcr genes could be detected with a lower limit of 102 cultured bacteria. This test was highly specific and sensitive, and generated no false-positive results. The assay was also conclusive when applied to feces and soil samples containing mcr-1-positive Escherichia coli, which could facilitate the screening of mcr genes not only in the bacteria, but also directly from the environment. This simple, rapid, sensitive, and specific multiplex assay will be useful for rapid screening of the colistin resistance in both clinical medicine and animal husbandry.

  14. Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum.

    Science.gov (United States)

    Anniballi, Fabrizio; Auricchio, Bruna; Delibato, Elisabetta; Antonacci, Monia; De Medici, Dario; Fenicia, Lucia

    2012-01-27

    Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Development of SYBR Green and TaqMan quantitative real-time PCR assays for hepatopancreatic parvovirus (HPV) infecting Penaeus monodon in India.

    Science.gov (United States)

    Yadav, Reena; Paria, Anutosh; Mankame, Smruti; Makesh, M; Chaudhari, Aparna; Rajendran, K V

    2015-12-01

    Hepatopancreatic parvovirus (HPV) infects Penaeus monodon and causes mortality in the larval stages. Further, it has been implicated in the growth retardation in cultured P. monodon. Though different geographical isolates of HPV show large sequence variations, a sensitive PCR assay specific to Indian isolate has not yet been reported. Here, we developed a sensitive SYBR Green-based and TaqMan real-time PCR for the detection and quantification of the virus. A 441-bp PCR amplicon was cloned in pTZ57 R/T vector and the plasmid copy number was estimated. A 10-fold serial dilution of the plasmid DNA from 1 × 10(9) copies to 1 copy was prepared and used as the standard. The primers were tested initially using the standard on a conventional PCR format to determine the linearity of detection. The standards were further tested on real-time PCR format using SYBR Green and TaqMan chemistry and standard curves were generated based on the Ct values from three well replicates for each dilution. The assays were found to be sensitive, specific and reproducible with a wide dynamic range (1 × 10(9) to 10 copies) with coefficient of regression (R(2)) > 0.99, calculated average slope -3.196 for SYBR Green assay whereas, for TaqMan assay it was >0.99 and -3.367, respectively. The intra- and inter-assay variance of the Ct values ranged from 0.26% to 0.94% and 0.12% to 0.81%, respectively, for SYBR Green assay, and the inter-assay variance of the Ct values for TaqMan assay ranged from 0.07% to 1.93%. The specificity of the assays was proved by testing other DNA viruses of shrimp such as WSSV, IHHNV and MBV. Standardized assays were further tested to detect and quantify HPV in the post-larvae of P. monodon. The result was further compared with conventional PCR to test the reproducibility of the test. The assay was also used to screen Litopeneaus vannamei, Macrobrachium rosenbergii and Scylla serrata for HPV. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

    OpenAIRE

    Paudel, Damodar; Jarman, Richard; Limkittikul, Kriengsak; Klungthong, Chonticha; Chamnanchanunt, Supat; Nisalak, Ananda; Gibbons, Robert; Chokejindachai, Watcharee

    2011-01-01

    Background : Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conven...

  17. Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis.

    Science.gov (United States)

    García, Alfredo; Martínez, Remigio; Benitez-Medina, José Manuel; Risco, David; Garcia, Waldo Luis; Rey, Joaquín; Alonso, Juan Manuel; Hermoso de Mendoza, Javier

    2013-01-01

    Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.

  18. Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis

    OpenAIRE

    García, Alfredo; Martínez, Remigio; Benitez-Medina, José Manuel; Risco, David; García, Waldo Luis; Rey, Joaquín; Alonso, Juan Manuel; de Mendoza, Javier Hermoso

    2013-01-01

    Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was ...

  19. Real-Time Detection and Identification of Chlamydophila Species in Veterinary Specimens by Using SYBR Green-Based PCR Assays

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Kabell, Susanne; Pedersen, Karl

    2011-01-01

    . felis was found in 10 cats, Cp. caviae was found in one guinea pig, and Cp. abortus was detected in one sheep. The screening assay appeared more sensitive than traditional microscopical examination of stained tissue smears. By combining a fast, robust, and cost-effective method for sample preparation...... or tissue specimens from heart, liver, and spleen without further purification. The assays were evaluated on veterinary specimens where all samples were screened using a family-specific PCR, and positive samples were further tested using species-specific PCRs. Cp. psittaci was detected in 47 birds, Cp...... with a highly sensitive family-specific PCR, we were able to screen for Chlamydiaceae in veterinary specimens and confirm the species in positive samples with additional PCR assays....

  20. Real-time PCR using SYBR Green for the detection of Shigella spp. in food and stool samples.

    Science.gov (United States)

    Mokhtari, W; Nsaibia, S; Gharbi, A; Aouni, M

    2013-02-01

    Shigella spp are exquisitely fastidious Gram negative organisms that frequently get missed in the detection by traditional culture methods. For this reason, this work has adapted a classical PCR for detection of Shigella in food and stool specimens to real-time PCR using the SYBR Green format. This method follows a melting curve analysis to be more rapid and provide both qualitative and quantitative data about the targeted pathogen. A total of 117 stool samples with diarrhea and 102 food samples were analyzed in Public Health Regional Laboratory of Nabeul by traditional culture methods and real-time PCR. To validate the real-time PCR assay, an experiment was conducted with both spiked and naturally contaminated stool samples. All Shigella strains tested were ipaH positive and all non-Shigella strains yielded no amplification products. The melting temperature (T(m) = 81.5 ± 0.5 °C) was consistently specific for the amplicon. Correlation coefficients of standard curves constructed using the quantification cycle (C(q)) versus copy numbers of Shigella showed good linearity (R² = 0.995; slope = 2.952) and the minimum level of detection was 1.5 × 10³ CFU/g feces. All food samples analyzed were negative for Shigella by standard culture methods, whereas ipaH was detected in 8.8% culture negative food products. Moreover, the ipaH specific PCR system increased the detection rate over that by culture alone from 1.7% to 11.1% among patients with diarrhea. The data presented here shows that the SYBR Green I was suitable for use in the real-time PCR assay, which provided a specific, sensitive and efficient method for the detection and quantification of Shigella spp in food and stool samples. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. A SYBR Green real-time PCR assay to detect and quantify pork meat in processed poultry meat products.

    Science.gov (United States)

    Soares, Sónia; Amaral, Joana S; Oliveira, M Beatriz P P; Mafra, Isabel

    2013-05-01

    Species identification in meat products has grown in interest in recent years since these foodstuffs are susceptible targets for fraudulent labelling. In this work, a real-time PCR approach based on SYBR Green dye was proposed for the quantitative detection of pork meat in processed meat products. For the development of the method, binary meat mixtures containing known amounts of pork meat in poultry meat were used to obtain a normalised calibration model from 0.1 to 25% with high linear correlation and PCR efficiency. The method revealed high specificity by melting curve analysis, being successfully validated through its application to blind meat mixtures, which confirmed its adequacy for pork meat determination. The fully applicability of the method was further demonstrated in commercial meat products, allowing verification of labelling compliance and identification of meat species in processed foods. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India

    Directory of Open Access Journals (Sweden)

    K P Dinoop

    2016-01-01

    Interpretation & conclusions: Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods.

  3. Differential Diagnosis of Entamoeba spp. in Clinical Stool Samples Using SYBR Green Real-Time Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Thiago dos Santos Gomes

    2014-01-01

    Full Text Available Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (Tm was 73°C and 70°C, respectively. For E. hartmanni, the Tm was 73°C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries.

  4. Differential diagnosis of Entamoeba spp. in clinical stool samples using SYBR green real-time polymerase chain reaction.

    Science.gov (United States)

    Gomes, Thiago Dos Santos; Garcia, Mariana Coimbra; de Souza Cunha, Flavia; Werneck de Macedo, Heloisa; Peralta, José Mauro; Peralta, Regina Helena Saramago

    2014-01-01

    Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (T(m)) was 73 °C and 70 °C, respectively. For E. hartmanni, the T(m) was 73 °C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries.

  5. Differential Diagnosis of Entamoeba spp. in Clinical Stool Samples Using SYBR Green Real-Time Polymerase Chain Reaction

    Science.gov (United States)

    Gomes, Thiago dos Santos; Garcia, Mariana Coimbra; de Souza Cunha, Flavia; Peralta, José Mauro; Peralta, Regina Helena Saramago

    2014-01-01

    Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (T m) was 73°C and 70°C, respectively. For E. hartmanni, the T m was 73°C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries. PMID:24693242

  6. Quantification of ochratoxin A-producing molds in food products by SYBR Green and TaqMan real-time PCR methods

    DEFF Research Database (Denmark)

    Rodríguez, Alicia; Rodríguez, Mar; Luque, M. Isabel

    2011-01-01

    Ochratoxin A (OTA) is a mycotoxin synthesized by a variety of different fungi, most of them from the genera Penicillium and Aspergillus. Early detection and quantification of OTA producing species is crucial to improve food safety. In the present work, two protocols of real-time qPCR based on SYBR...... 1 and 10 conidia/g for SYBR Green assay and TaqMan. No significant differences were found between the Ct values obtained from pure mold DNA and pure mold DNA mixed with food DNA. The ability of the designed qPCR methods to quantify two known conidial suspensions inoculated on several foods...... was evaluated. The amount of conidia assessed by both qPCR methods was close to the inoculated amount for most foods and indicates that the described procedure holds potential for use for the detection and quantification of OTA producing molds in foods....

  7. A SYBR®Green-based real-time PCR method for improved detection of mcr-1-mediated colistin resistance in human stool samples.

    Science.gov (United States)

    Donà, Valentina; Bernasconi, Odette J; Kasraian, Sara; Tinguely, Regula; Endimiani, Andrea

    2017-06-01

    The aim of this study was to design a rapid and sensitive real-time PCR (rt-PCR) method for colistin resistance mcr-1 gene detection in human faecal samples. Stools (n=88) from 36 volunteers were analysed. To isolate mcr-1-producing Enterobacteriaceae, samples were enriched overnight in Luria-Bertani (LB) broth containing 2mg/L colistin and were then plated on selective agar plates with 4mg/L colistin. A SYBR ® Green-based rt-PCR targeting mcr-1 was then designed. For method validation and to establish the limit of detection (LOD), total DNA was extracted from mcr-1-negative and mcr-1-positive Escherichia coli. rt-PCR was also performed with DNA extracted from 88 native stools and after enriching them in LB broth containing colistin. Based on the culture approach, three unique volunteers resulted colonised with mcr-1-harboring E. coli strains. For culture isolates, rt-PCR exhibited a LOD of 10 genomic copies/reaction, with both sensitivity and specificity of 100%. Nevertheless, when testing native stools, only two of the three mcr-1-positive specimens were detected. However, after enrichment in LB broth containing colistin, the rt-PCR was strongly positive for all culture-positive samples. The average cycle threshold was 22, granting rapid and confident detection of positive specimens within 30 cycles. No false positives were observed for the remaining 85 culture-negative specimens. A rapid rt-PCR for detection of mcr-1 from stool specimens was developed. The detection rate was increased by testing selective broth enrichments. This approach also has the advantage of concomitant isolation of mcr-1-harboring strains for further antimicrobial susceptibility and genetic testing. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  8. Development and validation of two SYBR green PCR assays and a multiplex real-time PCR for the detection of Shiga toxin-producing Escherichia coli in meat.

    Science.gov (United States)

    Brusa, Victoria; Galli, Lucía; Linares, Luciano H; Ortega, Emanuel E; Lirón, Juan P; Leotta, Gerardo A

    2015-12-01

    Shiga toxin-producing Escherichia coli (STEC) are recognized as food-borne pathogens. We developed and validated two SYBR green PCR (SYBR-PCR) and a real-time multiplex PCR (RT-PCR) to detect stx1 and stx2 genes in meat samples, and compared these techniques in ground beef samples from retail stores. One set of primers and one hydrolysis probe were designed for each stx gene. For RT-PCR, an internal amplification control (IAC) was used. All PCR intra-laboratory validations were performed using pure strains and artificially contaminated ground beef samples. A total of 50 STEC and 30 non-STEC strains were used. Naturally contaminated ground beef samples (n=103) were obtained from retail stores and screened with SYBR-PCR and RT-PCR, and stx-positive samples were processed for STEC isolation. In the intra-laboratory validation, each PCR obtained a 1×10(2) CFU mL(-1) limit of detection and 100% inclusivity and exclusivity. The same results were obtained when different laboratory analysts in alternate days performed the assay. The level of agreement obtained with SYBR-PCR and RT-PCR was kappa=0.758 and 0.801 (P<0.001) for stx1 and stx2 gene detection, respectively. Two PCR strategies were developed and validated, and excellent performance with artificially contaminated ground beef samples was obtained. However, the efforts made to isolate STEC from retail store samples were not enough. Only 11 STEC strains were isolated from 35 stx-positive ground beef samples identically detected by all PCRs. The combination of molecular approaches based on the identification of a virulence genotypic profile of STEC must be considered to improve isolation. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6 Otimização da PCR em tempo real - Sybr Green para detecção do Herpes Vírus Humano tipo 6 (HHV-6

    Directory of Open Access Journals (Sweden)

    Cynthia Liliane Motta do Canto

    2008-02-01

    Full Text Available HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL to 10-9 (equivalent to 2.46 molecules/µL. Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3' and by conventional nested PCR using primers HHV1 (outer: 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer: 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner and HHV4 (inner 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.HHV-6 é o agente etiológico do Exantema Súbito e considerado a sexta doença mais comum na infância. Em indivíduos imunocomprometidos, a reativação da infecção latente pode causar doença aguda ou morte. Padronizamos PCR em Tempo Real utilizando a química Sybr Green na detecção do HHV-6 e comparamos os resultados com a PCR convencional. Um fragmento de 214 pb foi clonado através do kit pGEM-T do sistema Promega. Com este clone, foram feitas diluições seriadas em um pool de leucócitos negativos a partir de 10-6 ng/µL (equivalente a 2465,8 moleculas/µL até 10-9 (equivalente a 2,46 moleculas/µL. As diluições foram amplificadas por PCR em Tempo Real utilizando Sybr Green, com

  10. Development of duplex SYBR Green I-based real-time quantitative reverse-transcription PCR for detection and discrimination of grapevine viruses

    Science.gov (United States)

    A SYBR® Green-based real-time quantitative reverse transcription PCR (qRT-PCR) assay in combination with melt curve analysis (MCA) was developed for the detection of nine grapevine viruses. The detection limits for singleplex qRT-PCR for all nine grapevine viruses were determined to be in the range ...

  11. An Undergraduate Laboratory Experiment for Upper-Level Forensic Science, Biochemistry, or Molecular Biology Courses: Human DNA Amplification Using STR Single Locus Primers by Real-Time PCR with SYBR Green Detection

    Science.gov (United States)

    Elkins, Kelly M.; Kadunc, Raelynn E.

    2012-01-01

    In this laboratory experiment, real-time polymerase chain reaction (real-time PCR) was conducted using published human TPOX single-locus DNA primers for validation and various student-designed short tandem repeat (STR) primers for Combined DNA Index System (CODIS) loci. SYBR Green was used to detect the amplification of the expected amplicons. The…

  12. Real-time PCR Detection of Brucella Abortus: A Comparative Study of SYBR Green I, 5'-exonuclease, and Hybridization Probe Assays

    Energy Technology Data Exchange (ETDEWEB)

    Newby, Deborah Trishelle; Hadfield, Ted; Roberto, Francisco Figueroa

    2003-08-01

    Real-time PCR provides a means of detecting and quantifying DNA targets by monitoring PCR product accumulation during cycling as indicated by increased fluorescence. A number of different approaches can be used to generate the fluorescence signal. Three approaches—SYBR Green I (a double-stranded DNA intercalating dye), 5'-exonuclease (enzymatically released fluors), and hybridization probes (fluorescence resonance energy transfer)—were evaluated for use in a real-time PCR assay to detect Brucella abortus. The three assays utilized the same amplification primers to produce an identical amplicon. This amplicon spans a region of the B. abortus genome that includes portions of the alkB gene and the IS711 insertion element. All three assays were of comparable sensitivity, providing a linear assay over 7 orders of magnitude (from 7.5 ng down to 7.5 fg). However, the greatest specificity was achieved with the hybridization probe assay.

  13. Quantification of hTERT Splice Variants in Melanoma by SYBR Green Real-time Polymerase Chain Reaction Indicates a Negative Regulatory Role for the β Deletion Variant

    Directory of Open Access Journals (Sweden)

    Lisa F. Lincz

    2008-10-01

    Full Text Available Telomerase activity is primarily determined by transcriptional regulation of the catalytic subunit, human telomerase reverse transcriptase (hTERT. Several mRNA splice variants for hTERT have been identified, but it is not clear if telomerase activity is determined by the absolute or relative levels of full-length (functional and variant hTERT transcripts. We have developed an SYBR green-based reverse transcription-quantitative polymerase chain reaction assay for the enumeration of the four common hTERT mRNA variants and correlated these with telomerase activity and telomere length in 24 human melanoma cell lines. All except five of the lines expressed four hTERT transcripts, with an overall significant level of co-occurrence between absolute mRNA levels of full-length α+/β+ hTERT and the three splice variants α-/β+, α+/β-, and α-/β-. On average, α+/β+ made up the majority (48.1% of transcripts, followed by α+/β- (44.6%, α-/β- (4.4%, and α-/β+ (2.9%. Telomerase activity ranged from 1 to 247 relative telomerase activity and correlated most strongly with the absolute amount of α+/β+ (R = 0.791, P = .000004 and the relative amount of α+/β- (R = -0.465, P = .022. This study shows that telomerase activity in melanoma cells is best determined by the absolute expression of full-length hTERT mRNA and indicates a role for the hTERT β deletion variant in the negative regulation of enzyme activity.

  14. Relative quantification and detection of different types of infectious bursal disease virus in bursa of Fabricius and cloacal swabs using real time RT-PCR SYBR green technology

    DEFF Research Database (Denmark)

    Li, Yiping; Handberg, K.J.; Kabell, Susanne

    2007-01-01

    In present study, different types of infectious bursal disease virus (IBDV), virulent strain DK01, classic strain F52/70 and vaccine strain D78 were quantified and detected in infected bursa of Fabricius (BF) and cloacal swabs using quantitative real time RT-PCR with SYBR green dye. For selection...... or F52/70 inoculation were detected as virus positive at day I post inoculation (p.i.). The D78 viral load peaked at day 4 and day 8 p.i., while the DK01 and F52/70 viral load showed relatively high levels at day 2 p.i. In cloacal swabs, viruses detectable were at day 2 p.i. for DK01 and F52/70, day 8...... of a suitable internal control gene, real time PCR parameters were evaluated for three candidate genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 28S rRNA and beta-actin to IBDVs. Based on this P-actin was selected as an internal control for quantification of IBDVs in BF. All BF samples with D78, DK01...

  15. Qualitative Sybr Green real-time detection of single nucleotide polymorphisms responsible for target-site resistance in insect pests: the example of Myzus persicae and Musca domestica.

    Science.gov (United States)

    Puggioni, V; Chiesa, O; Panini, M; Mazzoni, E

    2017-02-01

    Chemical insecticides have been widely used to control insect pests, leading to the selection of resistant populations. To date, several single nucleotide polymorphisms (SNPs) have already been associated with insecticide resistance, causing reduced sensitivity to many classes of products. Monitoring and detection of target-site resistance is currently one of the most important factors for insect pest management strategies. Several methods are available for this purpose: automated and high-throughput techniques (i.e. TaqMan or pyrosequencing) are very costly; cheaper alternatives (i.e. RFLP or PASA-PCRs) are time-consuming and limited by the necessity of a final visualization step. This work presents a new approach (QSGG, Qualitative Sybr Green Genotyping) which combines the specificity of PASA-PCR with the rapidity of real-time PCR analysis. The specific real-time detection of Cq values of wild-type or mutant alleles (amplified used allele-specific primers) allows the calculation of ΔCqW-M values and the consequent identification of the genotypes of unknown samples, on the basis of ranges previously defined with reference clones. The methodology is applied here to characterize mutations described in Myzus persicae and Musca domestica and we demonstrate it represents a valid, rapid and cost-effective technique that can be adopted for monitoring target-site resistance in field populations of these and other insect species.

  16. Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil.

    Science.gov (United States)

    Romeiro, Marilia Farignoli; Souza, William Marciel de; Tolardo, Aline Lavado; Vieira, Luiz Carlos; Colombo, Tatiana Elias; Aquino, Victor Hugo; Nogueira, Maurício Lacerda; Figueiredo, Luiz Tadeu Moraes

    2016-01-01

    The genus Flavivirus includes several pathogenic species that cause severe illness in humans. Therefore, a rapid and accurate molecular method for diagnosis and surveillance of these viruses would be of great importance. Here, we evaluate and optimize a quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) method for the diagnosis of the Flavivirus genus. We evaluated different commercial kits that use the SYBR Green system for real-time RT-PCR with a primer set that amplifies a fragment of the NS5 flavivirus gene. The specificity and sensitivity of the assay were tested using twelve flaviviruses and ribonucleic acid (RNA) transcribed from the yellow fever virus. Additionally, this assay was evaluated using the sera of 410 patients from different regions of Brazil with acute febrile illness and a negative diagnosis for the dengue virus. The real-time RT-PCR amplified all flaviviruses tested at a melting temperature of 79.92 to 83.49°C. A detection limit of 100 copies per ml was determined for this assay. Surprisingly, we detected dengue virus in 4.1% (17/410) of samples from patients with febrile illness and a supposedly negative dengue infection diagnosis. The viral load in patients ranged from 2.1×107to 3.4×103copies per ml. The real-time RT-PCR method may be very useful for preliminary diagnoses in screenings, outbreaks, and other surveillance studies. Moreover, this assay can be easily applied to monitor viral activity and to measure viral load in pathogenesis studies.

  17. Development of a SYBR Green real-time PCR assay with melting curve analysis for simultaneous detection and differentiation of canine adenovirus type 1 and type 2.

    Science.gov (United States)

    Balboni, Andrea; Dondi, Francesco; Prosperi, Santino; Battilani, Mara

    2015-09-15

    Canine adenovirus type 1 (CAdV-1) and canine adenovirus type 2 (CAdV-2) cause infectious canine hepatitis (ICH) and infectious tracheobronchitis (ITB) in dogs, respectively. Cases of ICH have been documented in recent years and recent surveys have demonstrated a wide percentage of asymptomatic CAdV-1 infection in the canine population. Since both CAdV types are detectable in the same biological matrices, and viral coinfection with CAdV-1 and CAdV-2 are reported with high frequency, it is urgent to have available a rapid, highly sensitive and specific assay for the diagnosis of CAdV infection and distinction between CAdV-1 and CAdV-2. In order to detect canine adenovirus in biological samples and to rapidly distinguish the two viral types, a SYBR Green real-time PCR assay was optimized to discriminate CAdV-1 and CAdV-2 via a melting curve analysis. The developed assay showed high sensitivity and reproducibility and was highly efficient and specific in discriminating the two CAdV types. This reliable and rapid technique may represent a simple, useful and economic option for simultaneous CAdV types detection, which would be feasible and attractive for all diagnostic laboratories, both for clinical purposes and for epidemiological investigations. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Development and validation of a SYBR Green real-time PCR assay for rapid and quantitative detection of goose interferons and proinflammatory cytokines.

    Science.gov (United States)

    Zhou, Hao; Chen, Shun; Qi, Yulin; Wang, Mingshu; Jia, Renyong; Zhu, Dekang; Liu, Mafeng; Liu, Fei; Chen, Xiaoyue; Cheng, Anchun

    2015-10-01

    Real time quantitative polymerase chain reaction (RT-qPCR) based on SYBR-Green I binding is a quick, reliable, and easy method for analyzing small amounts of mRNA. Viral pathogens are recognized at the time of infection by pattern recognition receptors; thus, the inflammatory cytokines (IL1β, IL6, and IL18) and antiviral cytokines (IFNα, IFNγ) are secreted by innate immune cells and induced to respond to the pathogens. The objective of this study was to develop an effective and sensitive RT-qPCR assay for the rapid and accurate quantification of goose cytokines: IFNα, IFNγ, IL1β, IL6, and IL18. Subsequently, the established methods were employed to detect the immune response in agonist-stimulated goose spleen cells in vitro. These data indicated that the established RT-qPCR is a reliable method for determining relative gene expression. The results revealed that Imiquimod led to the significant upregulation of goose IFNα (P pathogens. © 2015 Poultry Science Association Inc.

  19. Detection of Salmonella in Shellfish Using SYBR Green™ I-Based Real-Time Multiplexed PCR Assay Targeting invA and spvB

    KAUST Repository

    Gangwar, Maulshree

    2012-09-23

    A SYBR Green™ I-based real-time multiplexed PCR assay was developed targeting invA and spvB for the detection of Salmonella strains in shellfish after both hns and invA genes were identified in all Salmonella strains. Simultaneously, the 16S rRNA gene was used as a PCR internal amplification control (IAC). All 89 Salmonella strains tested in this study exhibited amplification of invA, whereas only 21 (23. 6 %) were PCR positive for spvB. The sensitivity of detection of all three targeted genes was 1 ng, which is equivalent to approximately 105 colony-forming unit (CFU) of Salmonella enterica. The analysis showed specific PCR products that were identified by reproducible melt temperature profiles (invA, 84. 27 ± 1. 7 °C; spvB, 88. 76 ± 1. 0 °C; and 16S rRNA gene, 87. 16 ± 0. 8 °C). The sensitivity of detection was 10 pg purified DNA (invA) or 105 CFU in 1 mL pure culture of S. enterica ATCC 14028. The above molecular detection method for Salmonella strains was successfully applied to the oyster homogenates (food matrix). An initial inoculum of 106 and 102 CFU Salmonella in 1 ml seeded oyster tissue homogenate was detected by multiplexed PCR for all three genes after 5 and 24 h of enrichment, respectively. Natural oysters isolated from Gulf of Mexico during the winter months exhibited negative PCR amplification results suggesting the absence of Salmonella. In contrast to conventional PCR, real-time multiplex PCR assay developed in this study is rapid and sensitive and will help Interstate Shellfish Sanitation Conference undertake appropriate measures to monitor Salmonella in oysters, thereby preventing disease outbreaks and consequently protecting consumer health. © 2012 Springer Science+Business Media, LLC.

  20. Optimization of rapid Salmonella enterica detection in liquid whole eggs by SYBR green I-based real-time reverse transcriptase-polymerase chain reaction.

    Science.gov (United States)

    Techathuvanan, Chayapa; D'Souza, Doris Helen

    2011-04-01

    Eggs and egg products have a high risk of Salmonella enterica serovar Enteritidis contamination leading to gastroenteritis outbreaks in humans. Thus, a rapid screening tool for viable Salmonella Enteritidis cells in the egg industry is needed. Our objective was to rapidly and sensitively detect viable Salmonella Enteritidis from spiked liquid whole eggs (LWEs) within 24 h using SYBR green I-based real-time reverse transcriptase-polymerase chain reaction (PCR) targeting the Salmonella specific invA gene along with an internal amplification control in a Bio-Rad iCycler. LWE was inoculated with Salmonella Enteritidis and mixed with tetrathionate broth, and 100 μL of serially diluted portions in phosphate-buffered saline was plated on Xylose Lysine Tergitol 4 agar or 5 mL were used for RNA extraction by the TRIzol method immediately or after enrichment of 6, 12, or 16 h at 37 °C. The real-time reverse transcriptase-PCR assay was carried out using previously described Salmonella invA gene primers. Melt temperature analysis of the PCR product was included to determine specific invA amplification. Without enrichment, the assay detection limit was 10(7) colony forming units (CFU)/25 mL LWE. After enrichment for 6 and 12 h, Salmonella Enteritidis could be detected from LWE up to 10(4) and 10(2) CFU/25 mL, respectively. Improved Salmonella Enteritidis detection up to 10(0) CFU/25 mL was obtained after 16-h enrichment. Even with 16-h enrichment, the results could be still be obtained within 24 h, which is much faster than by traditional cultural detection that takes several days. Therefore, this assay appears suitable for routine detection of Salmonella enterica contamination by the egg industry to help prevent the transmission of egg-associated Salmonella outbreaks and timely recall of contaminated products. © Mary Ann Liebert, Inc.

  1. APLIKASI KUANTIFIKASI KOI HERPESVIRUS : REAL TIME – QUANTITATIVE POLYMERASE CHAIN REACTION (RT-Q PCR MENGGUNAKAN SYBR GREEN PADA IKAN MAS (Cyprinus carpio

    Directory of Open Access Journals (Sweden)

    Isti Koesharyani

    2017-08-01

    Full Text Available Koi Herpes Virus (KHV di Indonesia sejak tahun 2002 merupakan penyakit mematikan yang menyerang ikan koi Cyprinus carpio koi dan ikan mas Cyprinus carpio carpio, dan sampai saat ini, infeksi KHV dilaporkan sudah menyebar hampir di seluruh dunia. Untuk mengetahui adanya infeksi KHV perlu cara diagnosa yang sangat akurat/sensitif, sehingga keberadaan KHV dapat diketahui secara pasti dengan tingkat sensitivitas yang lebih baik pada ikan budidaya. Tujuan dari penelitian ini adalah untuk mengaplikasikan teknik deteksi dengan real time quantitative polymerase chain reaction (RT- qPCR/qPCR guna mengetahui adanya infeksi KHV secara kuantitatif pada ikan mas dengan mengetahui kandungan virus (viral load. Sebanyak masing-masing 3 ekor sampel diperoleh dari sentra budidaya ikan mas di Cirata-Jawa Barat, Maninjau-Sumatera Barat, dan Banjarmasin-Kalimantan Selatan. Sampel-sampel tersebut selanjutnya dianalisa keberadaan KHV-nya dengan RT-qPCR menggunakan SYBR Green. Hasil pengujian menunjukkan bahwa jumlah tertinggi (viral load diperoleh dari ikan mas asal Cirata-3 dengan nilai Threshold Cycle (Ct. 18,24 atau setara dengan 3,4 x 107 kopi, dan terendah dari ikan mas asal Banjarmasin-3 dengan nilai Ct. 33,39 atau 1,8 x 102 kopi. Dua standar yang digunakan dalam pengujian ini berupa plasmid dengan jumlah kopi 2 x 104 (Ct 27,24 dan 2 x 103 (Ct 30,24 dan kontrol atau Non Template Control (NTC adalah 3,1 x 10 atau dengan nilai Ct 35,65. Uji aplikasi deteksi KHV dengan metode RT-qPCR ini memberikan hasil yang lebih sensitif, di mana sampel yang tidak terdeteksi dengan metode PCR konvensional dapat dideteksi dan dihitung jumlah kopi DNA (DNA copy. Since 2002, Koi herpesvirus (KHV in Indonesian has been a malignant diseases, now recognized as a worldwide cause of mortality among populations of koi Cyprinus carpio koi and common carp Cyprinus carpio carpio. To determine the presence of infection is required the KHV diagnosis method with highly accurate and sensitive

  2. SYBR Green real-time PCR-RFLP assay targeting the plasmodium cytochrome B gene--a highly sensitive molecular tool for malaria parasite detection and species determination.

    Directory of Open Access Journals (Sweden)

    Weiping Xu

    Full Text Available A prerequisite for reliable detection of low-density Plasmodium infections in malaria pre-elimination settings is the availability of ultra-sensitive and high-throughput molecular tools. We developed a SYBR Green real-time PCR restriction fragment length polymorphism assay (cytb-qPCR targeting the cytochrome b gene of the four major human Plasmodium species (P. falciparum, P. vivax, P. malariae, and P. ovale for parasite detection and species determination with DNA extracted from dried blood spots collected on filter paper. The performance of cytb-qPCR was first compared against four reference PCR methods using serially diluted Plasmodium samples. The detection limit of the cytb-qPCR was 1 parasite/μl (p/μl for P. falciparum and P. ovale, and 2 p/μl for P. vivax and P. malariae, while the reference PCRs had detection limits of 0.5-10 p/μl. The ability of the PCR methods to detect low-density Plasmodium infections was then assessed using 2977 filter paper samples collected during a cross-sectional survey in Zanzibar, a malaria pre-elimination setting in sub-Saharan Africa. Field samples were defined as 'final positive' if positive in at least two of the five PCR methods. Cytb-qPCR preformed equal to or better than the reference PCRs with a sensitivity of 100% (65/65; 95%CI 94.5-100% and a specificity of 99.9% (2910/2912; 95%CI 99.7-100% when compared against 'final positive' samples. The results indicate that the cytb-qPCR may represent an opportunity for improved molecular surveillance of low-density Plasmodium infections in malaria pre-elimination settings.

  3. SYBR Green real-time PCR-RFLP assay targeting the plasmodium cytochrome B gene--a highly sensitive molecular tool for malaria parasite detection and species determination.

    Science.gov (United States)

    Xu, Weiping; Morris, Ulrika; Aydin-Schmidt, Berit; Msellem, Mwinyi I; Shakely, Delér; Petzold, Max; Björkman, Anders; Mårtensson, Andreas

    2015-01-01

    A prerequisite for reliable detection of low-density Plasmodium infections in malaria pre-elimination settings is the availability of ultra-sensitive and high-throughput molecular tools. We developed a SYBR Green real-time PCR restriction fragment length polymorphism assay (cytb-qPCR) targeting the cytochrome b gene of the four major human Plasmodium species (P. falciparum, P. vivax, P. malariae, and P. ovale) for parasite detection and species determination with DNA extracted from dried blood spots collected on filter paper. The performance of cytb-qPCR was first compared against four reference PCR methods using serially diluted Plasmodium samples. The detection limit of the cytb-qPCR was 1 parasite/μl (p/μl) for P. falciparum and P. ovale, and 2 p/μl for P. vivax and P. malariae, while the reference PCRs had detection limits of 0.5-10 p/μl. The ability of the PCR methods to detect low-density Plasmodium infections was then assessed using 2977 filter paper samples collected during a cross-sectional survey in Zanzibar, a malaria pre-elimination setting in sub-Saharan Africa. Field samples were defined as 'final positive' if positive in at least two of the five PCR methods. Cytb-qPCR preformed equal to or better than the reference PCRs with a sensitivity of 100% (65/65; 95%CI 94.5-100%) and a specificity of 99.9% (2910/2912; 95%CI 99.7-100%) when compared against 'final positive' samples. The results indicate that the cytb-qPCR may represent an opportunity for improved molecular surveillance of low-density Plasmodium infections in malaria pre-elimination settings.

  4. SYBR Green Real-Time PCR-RFLP Assay Targeting the Plasmodium Cytochrome B Gene – A Highly Sensitive Molecular Tool for Malaria Parasite Detection and Species Determination

    Science.gov (United States)

    Xu, Weiping; Morris, Ulrika; Aydin-Schmidt, Berit; Msellem, Mwinyi I.; Shakely, Delér; Petzold, Max; Björkman, Anders; Mårtensson, Andreas

    2015-01-01

    A prerequisite for reliable detection of low-density Plasmodium infections in malaria pre-elimination settings is the availability of ultra-sensitive and high-throughput molecular tools. We developed a SYBR Green real-time PCR restriction fragment length polymorphism assay (cytb-qPCR) targeting the cytochrome b gene of the four major human Plasmodium species (P. falciparum, P. vivax, P. malariae, and P. ovale) for parasite detection and species determination with DNA extracted from dried blood spots collected on filter paper. The performance of cytb-qPCR was first compared against four reference PCR methods using serially diluted Plasmodium samples. The detection limit of the cytb-qPCR was 1 parasite/μl (p/μl) for P. falciparum and P. ovale, and 2 p/μl for P. vivax and P. malariae, while the reference PCRs had detection limits of 0.5–10 p/μl. The ability of the PCR methods to detect low-density Plasmodium infections was then assessed using 2977 filter paper samples collected during a cross-sectional survey in Zanzibar, a malaria pre-elimination setting in sub-Saharan Africa. Field samples were defined as ‘final positive’ if positive in at least two of the five PCR methods. Cytb-qPCR preformed equal to or better than the reference PCRs with a sensitivity of 100% (65/65; 95%CI 94.5–100%) and a specificity of 99.9% (2910/2912; 95%CI 99.7–100%) when compared against ‘final positive’ samples. The results indicate that the cytb-qPCR may represent an opportunity for improved molecular surveillance of low-density Plasmodium infections in malaria pre-elimination settings. PMID:25774805

  5. Diagnosis of Barmah Forest Virus Infection by a Nested Real-Time SYBR Green RT-PCR Assay

    OpenAIRE

    Hueston, Linda; Toi, Cheryl S.; Jeoffreys, Neisha; Sorrell, Tania; Gilbert, Gwendolyn

    2013-01-01

    Barmah Forest virus (BFV) is a mosquito borne (+) ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR ne...

  6. Development of one-step SYBR Green real-time RT-PCR for quantifying bovine viral diarrhea virus type-1 and its comparison with conventional RT-PCR

    Directory of Open Access Journals (Sweden)

    Lou Sai

    2011-07-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV is a worldwide pathogen in cattle and acts as a surrogate model for hepatitis C virus (HCV. One-step real-time fluorogenic quantitative reverse transcription polymerase chain reaction (RT-PCR assay based on SYBR Green I dye has not been established for BVDV detection. This study aims to develop a quantitative one-step RT-PCR assay to detect BVDV type-1 in cell culture. Results One-step quantitative SYBR Green I RT-PCR was developed by amplifying cDNA template from viral RNA and using in vitro transcribed BVDV RNA to establish a standard curve. The assay had a detection limit as low as 100 copies/ml of BVDV RNA, a reaction efficiency of 103.2%, a correlation coefficient (R2 of 0.995, and a maximum intra-assay CV of 2.63%. It was 10-fold more sensitive than conventional RT-PCR and can quantitatively detect BVDV RNA levels from 10-fold serial dilutions of titrated viruses containing a titer from 10-1 to 10-5 TCID50, without non-specific amplification. Melting curve analysis showed no primer-dimers and non-specific products. Conclusions The one-step SYBR Green I RT-PCR is specific, sensitive and reproducible for the quantification of BVDV in cell culture. This one-step SYBR Green I RT-PCR strategy may be further optimized as a reliable assay for diagnosing and monitoring BVDV infection in animals. It may also be applied to evaluate candidate agents against HCV using BVDV cell culture model.

  7. Diagnosis of Barmah Forest virus infection by a nested real-time SYBR green RT-PCR assay.

    Directory of Open Access Journals (Sweden)

    Linda Hueston

    Full Text Available Barmah Forest virus (BFV is a mosquito borne (+ ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR negative. Two culture-positive (neutralizing antibody negative samples were positive on first round PCR, while one sample (IgM and neutralizing antibody strongly positive, IgG negative was positive on second round PCR, suggesting that viral RNA is detectable and transiently present in early infection. PCR can provide results faster than culture, is capable of high throughput and by sequencing the PCR product strain variants can be characterized.

  8. Comparison and evaluation of conventional RT-PCR, SYBR green I and TaqMan real-time RT-PCR assays for the detection of porcine epidemic diarrhea virus.

    Science.gov (United States)

    Zhou, Xinrong; Zhang, Tiansheng; Song, Deping; Huang, Tao; Peng, Qi; Chen, Yanjun; Li, Anqi; Zhang, Fanfan; Wu, Qiong; Ye, Yu; Tang, Yuxin

    2017-06-01

    Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal disease, resulting in substantial economic losses to the swine industry worldwide. In this study, three assays, namely a conventional reverse transcription-polymerase chain reaction (RT-PCR), a SYBR Green I real-time RT-PCR and a TaqMan real-time RT-PCR targeting the highly conserved M gene of PEDV, were developed and evaluated. Then, the analytical specificity, sensitivity and reproducibility of these assays were determined and compared. The TaqMan real-time RT-PCR was 100-fold and 10,000-fold more sensitive than that of the SYBR Green I real-time RT-PCR and the conventional RT-PCR, respectively. The analytical sensitivity of TaqMan real-time RT-PCR was 10 copies/μl of target gene and no cross amplification with other viruses tested was observed. With the features of high specificity, sensitivity, and reproducibility, the TaqMan real-time RT-PCR established in this study could be a useful tool for clinical diagnosis, epidemiological surveys and outbreak investigations of PED. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Touch-down reverse transcriptase-PCR detection of IgV(H) rearrangement and Sybr-Green-based real-time RT-PCR quantitation of minimal residual disease in patients with chronic lymphocytic leukemia

    Czech Academy of Sciences Publication Activity Database

    Peková, Soňa; Marková, J.; Pajer, Petr; Dvořák, Michal; Cetkovský, P.; Schwarz, J.

    2005-01-01

    Roč. 9, č. 1 (2005), s. 23-34 ISSN 1084-8592 Institutional research plan: CEZ:AV0Z50520514 Keywords : minimal residual disease * chronic lymphocytic leukaemia * IgV(H) rearrangement Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.562, year: 2003

  10. Identification by real-time PCR with SYBR Green of Leishmania spp. and Serratia marcescens in canine 'sterile' cutaneous nodular lesions.

    Science.gov (United States)

    Cornegliani, Luisa; Corona, Antonio; Vercelli, Antonella; Roccabianca, Paola

    2015-06-01

    Noninfectious, non-neoplastic, nodular to diffuse, so-called 'sterile' granulomatous/pyogranulomatous skin lesions (SGPSLs) are infrequently identified in dogs and may represent a diagnostic challenge. Their correct identification is based on history, histopathology and absence of intralesional foreign bodies and micro-organisms. The aim of this study was to investigate the presence of Leishmania spp., Mycobacterium spp., Serratia marcescens and Nocardia spp. by real-time PCR in canine nodular skin lesions histologically diagnosed as putatively sterile. Formalin-fixed skin biopsies were collected from 40 dogs. All samples were associated with an SGPSL diagnosis characterized by multifocal, nodular to diffuse, periadnexal and perifollicular pyogranulomas/granulomas. Neither micro-organisms nor foreign bodies were detected with haematoxylin and eosin staining, under polarized light. Further analyses included periodic acid Schiff, Ziehl-Neelsen, Fite Faraco, Giemsa and Gram histochemical stains; anti-Bacillus Calmette-Guérin (BCG) and Leishmania spp. immunohistochemistry; and real-time PCR analysis for Leishmania spp., Mycobacterium spp., S. marcescens and Nocardia spp. Special stains and BCG/immunohistochemistry were negative in all samples. Real-time PCR was positive for Leishmania spp. in four of 40 biopsies and for S. marcescens in two of 40 samples. Real-time PCR for Mycobacterium spp. and Nocardia spp. was negative. No correlation between real-time PCR positivity and a specific histological pattern was identified. Leishmania spp. have been previously identified as possible agents of certain SGPSLs, while the involvement of S. marcescens has not been investigated previously. According to our findings, Serratia spp. should be included in the list of agents possibly associated with a subgroup of granulomatous/pyogranulomatous skin lesions in dogs. © 2015 ESVD and ACVD.

  11. Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals.

    Science.gov (United States)

    Zemtsova, Galina E; Montgomery, Merrill; Levin, Michael L

    2015-01-01

    Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays.

  12. A novel strategy for human papillomavirus detection and genotyping with SybrGreen and molecular beacon polymerase chain reaction.

    Science.gov (United States)

    Szuhai, K; Sandhaus, E; Kolkman-Uljee, S M; Lemaître, M; Truffert, J C; Dirks, R W; Tanke, H J; Fleuren, G J; Schuuring, E; Raap, A K

    2001-11-01

    Human papillomaviruses (HPVs) play an important role in the pathogenesis of cervical cancer. For identification of the large number of different HPV types found in (pre)malignant lesions, a robust methodology is needed that combines general HPV detection with HPV genotyping. We have developed for formaldehyde-fixed samples a strategy that, in a homogeneous, real-time fluorescence polymerase chain reaction (PCR)-based assay, accomplishes general HPV detection by SybrGreen reporting of HPV-DNA amplicons, and genotyping of seven prevalent HPV types (HPV-6, -11, -16, -18, -31, -33, -45) by real-time molecular beacon PCR. The false-positive rate of the HPV SybrGreen-PCR was 4%, making it well suited as a prescreening, general HPV detection technology. The type specificity of the seven selected HPV molecular beacons was 100% and double infections were readily identified. The multiplexing capacity of the HPV molecular beacon PCR was analyzed and up to three differently labeled molecular beacons could be used in one PCR reaction without observing cross talk. The inherent quantitation capacities of real-time fluorescence PCR allowed the determination of average HPV copy number per cell. We conclude that the HPV SybrGreen-PCR in combination with the HPV molecular beacon PCR provides a robust, sensitive, and quantitative general HPV detection and genotyping methodology.

  13. Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR

    DEFF Research Database (Denmark)

    Hillig, Ann-Britt Nygaard; Jørgensen, Claus Bøttcher; Cirera, Susanna

    2007-01-01

    Background: Real-time quantitative PCR (qPCR) is a method for rapid and reliable quantification of mRNA transcription. Internal standards such as reference genes are used to normalise mRNA levels between different samples for an exact comparison of mRNA transcription level. Selection of high...... quality reference genes is of crucial importance for the interpretation of data generated by real-time qPCR. Results: In this study nine commonly used reference genes were investigated in 17 different pig tissues using real-time qPCR with SYBR green. The genes included beta-actin (ACTB), beta-2...

  14. Flow cytometric analysis using SYBR Green I for genome size estimation in coffee.

    Science.gov (United States)

    Ronildo Clarindo, Wellington; Roberto Carvalho, Carlos

    2011-02-01

    Plant genome size has been measured by flow cytometry using propidium iodide as a dye for nuclear DNA staining. However, some authors have reported the occurrence of genome size estimation errors, especially in plants rich in secondary metabolites, such as the coffee tree. In this context, we tested an alternative cytometric protocol using the SYBR Green I as a fluorochrome for stoichiometrically staining nuclear double-stranded DNA in Coffea canephora (2x) and Coffea arabica (4x). The results showed that the respective mean genome size measured from nuclei stained with SYBR Green I and propidium iodide was statistically identical. However, the G(0)/G(1) peaks of nuclei stained with SYBR Green I exhibited lower coefficient variations (1.57-2.85%) compared to those stained with propidium iodide (2.75-4.80%). Coefficient variation statistical data suggest that SYBR Green I is adequate for stoichiometric nuclei staining using this methodology. Our results provide evidence that SYBR Green I can be used in flow cytometry measurements of plants, with the advantages of minimizing errors in nuclear DNA content quantification, staining relatively quicker, with high affinity, and being less mutagenic than propidium iodide. Copyright © 2009 Elsevier GmbH. All rights reserved.

  15. SYBR safeTMefficiently replaces ethidium bromide in Aspergillus fumigatus gene disruption.

    Science.gov (United States)

    Canela, H M S; Takami, L A; Ferreira, M E S

    2017-02-08

    Invasive aspergillosis is a disease responsible for high mortality rates, caused mainly by Aspergillus fumigatus. The available drugs are limited and this disease continues to occur at an unacceptable frequency. Gene disruption is essential in the search for new drug targets. An efficient protocol for A. fumigatus gene disruption was described but it requires ethidium bromide, a genotoxic agent, for DNA staining. Therefore, the present study tested SYBR safe TM , a non-genotoxic DNA stain, in A. fumigatus gene disruption protocol. The chosen gene was cipC, which has already been disrupted successfully in our laboratory. A deletion cassette was constructed in Saccharomyces cerevisiae and used in A. fumigatus transformation. There was no statistical difference between the tested DNA stains. The success rate of S. cerevisiae transformation was 63.3% for ethidium bromide and 70% for SYBR safe TM . For A. fumigatus gene disruption, the success rate for ethidium bromide was 100 and 97% for SYBR safe TM . In conclusion, SYBR safe TM efficiently replaced ethidium bromide, making this dye a safe and efficient alternative for DNA staining in A. fumigatus gene disruption.

  16. The development of a rapid SYBR Green I-based quantitative PCR for detection of Duck circovirus.

    Science.gov (United States)

    Wan, Chunhe; Huang, Yu; Cheng, Longfei; Fu, Guanghua; Shi, Shao-hua; Chen, Hongmei; Peng, Chunxiang; Lin, Fang; Lin, Jiansheng

    2011-10-07

    This report describes a one-step real-time polymerase chain reaction assay based on SYBR Green I for detection of a broad range of duck circovirus (DuCV). Align with all DuCV complete genome sequences and other Genus Circovirus download from the GenBank (such as goose circovirus, pigeon circovirus), the primers targets to the replicate gene of DuCV were designed. The detection assay was linear in the range of 1.31 × 102-1.31 × 107 copies/μL. The reaction efficiency of the assay using the slope (the slope was -3.349) and the Y-intercept was 37.01 from the linear equation was estimated to be 0.99 and the correlation coefficient (R2) was 0.993. A series of experiments were carried out to assess the reproducibility, sensitivity, and specificity of the assay, following by the low intra-assay and inter-assay CVs for CT values obtained with the standard plasmids. The intra-assay CVs were equal or less than 1.89% and the inter-assay CVs were equal or less than 1.26%. There was no cross-reaction occurred with nucleic acids extracted from RA (Riemerella anatipestifer), E. coli (Escherichia coli), Duck Cholera (Pasteurella multocida), Avian influenza virus, avian paramyxovirus, Muscovy duck parvovirus, Duck reovirus, Duck hepatitis A virus as control templates. The nucleic acids extracted from samples of healthy ducks were used as negative controls. The assay was specific and reproducible. The established real time PCR was used to detect 45 DuCV-negative samples, which were tested using conventional PCR under the developed optimal conditions, each 15 for embryonated eggs, non-embryonated budgerigar eggs, newly hatched duck, the mixture of the lung, liver, spleen which were analysis for the presence of DuCV DNA, to conform that whether the DuCV can be transmitted vertically. Meanwhile, no positive result was shown by the real-time PCR method. The SYBR Green I-based quantitative PCR can therefore be practically used as an alternative diagnostic tool and a screening method

  17. 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.

  18. Comparison of multiple DNA dyes for real-time PCR: effects of dye concentration and sequence composition on DNA amplification and melting temperature

    DEFF Research Database (Denmark)

    Guðnason, Haukur; Dufva, Hans Martin; Bang, Dang Duong

    2007-01-01

    The importance of real-time polymerase chain reaction (PCR) has increased steadily in clinical applications over the last decade. Many applications utilize SYBR Green I dye to follow the accumulation of amplicons in real time. SYBR Green I has, however, a number of limitations that include...... investigate 15 different intercalating DNA dyes for their inhibitory effects on PCR, effects on DNA melting temperature and possible preferential binding to GC-rich sequences. Our results demonstrated that in contrast to the results of SYBR Green I, two intercalating dyes SYTO-13 and SYTO-82 do not inhibit...

  19. A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of 'Candidatus Phytoplasma mali'.

    Science.gov (United States)

    Aldaghi, M; Massart, S; Dutrecq, O; Bertaccini, A; Jijakli, M H; Lepoivre, P

    2009-03-01

    Different PCR protocols have been established for detection of European fruit trees phytoplasmas; however the majority of the procedures for extracting phytoplasma DNA are complex, time consuming, and expensive, with a risk of contamination or loss of target DNA. In present study, a crude extract preparation method previously used to detect other plant pathogens was adapted to samples from apple trees infected by 'Candidatus Phytoplasma mali'. End-point and real-time PCR detection of 'Ca. P. mali' were used to compare this extraction procedure with an established method for efficient extraction of purified DNA. The crude extract proved fully adequate for phytoplasma detection in samples from 86 in vitro and 35 in vivo apple shoots or plants and 10 periwinkle plants. High inter- and intra-run reproducibility was obtained for phytoplasma detection with different TaqMan MGB- or SYBR Green-based real-time PCR protocols applied to the crude extracts. Real-time PCR applied to serially diluted crude and purified extracts revealed the same phytoplasma detection limit (dilution up to 10(5)). All results confirm the suitability of this simple, quick, efficient extraction technique for accurate detection of 'Ca. P. mali' in different types of apple and periwinkle samples.

  20. Comparison of Parasite Burden Using Real-Time Polymerase Chain Reaction Assay and Limiting Dilution Assay in Leishma-nia major Infected Mouse

    Directory of Open Access Journals (Sweden)

    Somayeh GHOTLOO

    2015-12-01

    Full Text Available Background:Limiting dilution assay is considered as the gold standard method for quantifying the number of parasites in the animal model of Leishmania infection. Nowadays, real-time PCR is being increasingly applied to quantify infectious agents. In the present study, a real-time PCR assay was developed to estimate para­site burdens in lymph nodes of Leishmania major infected BALB/C mice. Enumera­tion of parasites was also performed by limiting dilution assay and compared with the results of real-time PCR based quantification.Methods:The SYBR Green based real- time PCR assay was performed to amplify a 75 bp fragment of superoxide dismutase B1 gene in the lymph nodes of L. major infected BALB/C mice 8 weeks post infection. Mice were infected subcutaneously at the base of their tail with 2 × 105L. major promastigotes in the stationary phase of growth. To compare parasite burdens obtained by real-time PCR assay with those of limiting dilution assay, twelve 8-fold serial dilutions of the lymph node homoge­nates were prepared in the Schneider medium and incubated at 26°C.After 7 days, wells containing motile parasites were identified by direct observation under an inverted light microscope and the total number of parasites was estimated using the ELIDA software.Results:Spearman's correlation coefficient of the parasite burdens between real-time PCR and limiting dilution assay was 0.72 (Pvalue = 0.008.Conclusion:Real-time PCR assay is an appropriate replacement to existing limit­ing dilution assay in quantifying parasite burden in the experimental model of Leishma­nia infection.

  1. Hot electron-induced electrogenerated chemiluminescence of SYBR[reg] Green I

    International Nuclear Information System (INIS)

    Laakso, Pirita; Anttila, Hannamari; Kairisto, Veli; Eskola, Jarkko; Kulmala, Sakari; Ala-Kleme, Timo

    2005-01-01

    Hot electrons can be injected from conductor/insulator/electrolyte (C/I/E) junctions into an aqueous electrolyte solution. Injected hot electrons induce electrogenerated chemiluminescence (ECL) of various luminophores in fully aqueous solutions. Such ECL gives the basis of electrochemiluminoimmunoassays and DNA-probe assays where different luminophores can be used as electrochemiluminescent labels. This work shows that SYBR[reg] Green I is suitable as an ECL label in detection methods based on C/I/E tunnel-emission electrodes such as oxide-coated aluminium, magnesium and silicon electrodes

  2. Development and validation of real-time PCR for rapid detection of Mecistocirrus digitatus.

    Directory of Open Access Journals (Sweden)

    Subhra Subhadra

    Full Text Available Hematophagous activity of Mecistocirrus digitatus, which causes substantial blood and weight loss in large ruminants, is an emerging challenge due to the economic loss it brings to the livestock industry. Infected animals are treated with anthelmintic drugs, based on the identification of helminth species and the severity of infection; however, traditional methods such as microscopic identification and the counting of eggs for diagnosis and determination of level of infection are laborious, cumbersome and unreliable. To facilitate the detection of this parasite, a SYBR green-based real-time PCR was standardized and validated for the detection of M. digitatus infection in cattle and buffaloes. Oligonucleotides were designed to amplify partial Internal Transcribed Spacer (ITS-1 sequence of M. digitatus. The specificity of the primers was confirmed by non-amplification of DNA extracted from other commonly occurring gastrointestinal nematodes in ruminants. Plasmids were ligated with partial ITS-1 sequence of M. digitatus, serially diluted (hundred fold and used as standards in the real-time PCR assay. The quantification cycle (Cq values were plotted against the standard DNA concentration to produce a standard curve. The assay was sensitive enough to detect one plasmid containing the M. digitatus DNA. Clinical application of this assay was validated by testing the DNA extracted from the faeces of naturally infected cattle (n = 40 and buffaloes (n = 25. The results were compared with our standard curve to calculate the quantity of M. digitatus in each faecal sample. The Cq value of the assay depicted a strong linear relationship with faecal DNA content, with a regression coefficient of 0.984 and efficiency of 99%. This assay has noteworthy advantages over the conventional methods of diagnosis because it is more specific, sensitive and reliable.

  3. Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data

    NARCIS (Netherlands)

    Ramakers, Christian; Ruijter, Jan M.; Deprez, Ronald H. Lekanne; Moorman, Antoon F. M.

    2003-01-01

    Quantification of mRNAs using real-time polymerase chain reaction (PCR) by monitoring the product formation with the fluorescent dye SYBR Green I is being extensively used in neurosciences, developmental biology, and medical diagnostics. Most PCR data analysis procedures assume that the PCR

  4. Quantification of M13 and T7 bacteriophages by TaqMan and SYBR green qPCR.

    Science.gov (United States)

    Peng, Xiujuan; Nguyen, Alex; Ghosh, Debadyuti

    2018-02-01

    TaqMan and SYBR Green quantitative PCR (qPCR) methods were developed as DNA-based approaches to reproducibly enumerate M13 and T7 phages from phage display selection experiments individually and simultaneously. The genome copies of M13 and T7 phages were quantified by TaqMan or SYBR Green qPCR referenced against M13 and T7 DNA standard curves of known concentrations. TaqMan qPCR was capable of quantifying M13 and T7 phage DNA simultaneously with a detection range of 2.75*10 1 -2.75*10 8 genome copies(gc)/μL and 2.66*10 1 -2.66*10 8 genome copies(gc)/μL respectively. TaqMan qPCR demonstrated an efficient amplification efficiency (E s ) of 0.97 and 0.90 for M13 and T7 phage DNA, respectively. SYBR Green qPCR was ten-fold more sensitive than TaqMan qPCR, able to quantify 2.75-2.75*10 7 gc/μL and 2.66*10 1 -2.66*10 7 gc/μL of M13 and T7 phage DNA, with an amplification efficiency E s of 1.06 and 0.78, respectively. Due to its superior sensitivity, SYBR Green qPCR was used to enumerate M13 and T7 phage display clones selected against a cell line, and quantified titers demonstrated accuracy comparable to titers from traditional double-layer plaque assay. Compared to enzyme linked immunosorbent assay, both qPCR methods exhibited increased detection sensitivity and reproducibility. These qPCR methods are reproducible, sensitive, and time-saving to determine their titers and to quantify a large number of phage samples individually or simultaneously, thus avoiding the need for time-intensive double-layer plaque assay. These findings highlight the attractiveness of qPCR for phage enumeration for applications ranging from selection to next-generation sequencing (NGS). Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Molecular detection of Phytophthora ramorum by real-time PCR using Taqman, SYBR Green and molecular beacons with three genes

    Science.gov (United States)

    G.J. Bilodeau; C.A. Lévesque; A.W.A.M. De Cock; C. Duchaine; G. Kristjansson; R.C. Hamelin

    2006-01-01

    Sudden oak death, caused by Phytophthora ramorum, is a severe disease that can affect numerous species of trees and shrubs. This pathogen has been spread via nursery stock, and quarantine measures are currently in place to prevent further spread. Molecular assays have been developed to rapidly detect and identify P. ramorum, but...

  6. A new whole mitochondrial genome qPCR (WMG-qPCR) with SYBR Green®to identify phlebotomine sand fly blood meals.

    Science.gov (United States)

    Rodrigues, Ana Caroline Moura; Magalhães, Rafaela Damasceno; Romcy, Kalil Andrade Mubarac; Freitas, Jeferson Lucas Sousa; Melo, Ana Carolina Fonseca Lindoso; Rodon, Fernanda Cristina Macedo; Bevilaqua, Claudia Maria Leal; Melo, Luciana Magalhães

    2017-04-30

    Phlebotomine sand flies are blood-feeding insects of marked medical and veterinary significance. Investigations on the biology of these insects hold great importance for both ecological and epidemiological purposes. The present work describes a new approach for real-time PCR (qPCR) with SYBR Green ® , named WMG-qPCR, to identify phlebotomine blood meals. The novelty of the assay was to design primers based on the Whole Mitochondrial Genome (WMG) of the potential hosts (human, dog, cat, brown rat and chicken) aiming to amplify through qPCR the regions of mitochondrial DNA (mtDNA) which are less conserved among all species. Initially, the best method for mtDNA extraction to be applied in WMG-qPCR was determined. Afterwards, amplification specificities were accessed by cross-reaction assays with mtDNA samples from all animal species, besides phlebotomine DNA. Finally, the selected primers were also tested for their limit of DNA detection through standard curves constructed by serial dilution of blood DNA obtained for each target animal species. The WMG-qPCR was able to detect as low as 10pL of blood, equivalent to 26, 84, 130, and 320fg DNA of cat, human, dog and rat, respectively. The assay was also capable to amplify as low as 5pL of chicken blood (5pg DNA). In conclusion, WMG-qPCR seems to be a promising tool to identify phlebotomine blood meals, with high species-specificity and sensitivity. Furthermore, as no supplementary techniques are required, this new approach presents minimized costs and simplified technical-training requirements for execution. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Effect of acidic pH on flow cytometric detection of bacteria stained with SYBR Green I and their distinction from background

    International Nuclear Information System (INIS)

    Baldock, Daniel; Nocker, Andreas; Nebe-von-Caron, Gerhard; Bongaerts, Roy

    2013-01-01

    Unspecific background caused by biotic or abiotic particles, cellular debris, or autofluorescence is a well-known interfering parameter when applying flow cytometry to the detection of microorganisms in combination with fluorescent dyes. We present here an attempt to suppress the background signal intensity and thus to improve the detection of microorganisms using the nucleic acid stain SYBR ® Green I. It has been observed that the fluorescent signals from SYBR Green I are greatly reduced at acidic pH. When lowering the pH of pre-stained samples directly prior to flow cytometric analysis, we hypothesized that the signals from particles and cells with membrane damage might therefore be reduced. Signals from intact cells, temporarily maintaining a neutral cytosolic pH, should not be affected. We show here that this principle holds true for lowering background interference, whereas the signals of membrane-compromised dead cells are only affected weakly. Signals from intact live cells at low pH were mostly comparable to signals without acidification. Although this study was solely performed with SYBR ® Green I, the principle of low pH flow cytometry (low pH-FCM) might hold promise when analyzing complex matrices with an abundance of non-cellular matter, especially when expanded to non-DNA binding dyes with a stronger pH dependence of fluorescence than SYBR Green I and a higher pK a value. (paper)

  8. Sensitivity of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood

    OpenAIRE

    da Costa Lima, Manoel Sebastião; Zorzenon, Denielly Christina Rodrigues; Dorval, Maria Elizabeth Cavalheiros; Pontes, Elenir Rose Jardim Cury; Oshiro, Elisa Teruya; Cunha, Rodrigo; Andreotti, Renato; Matos, Maria de Fatima Cepa

    2013-01-01

    Objective: To evaluate the effectiveness of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood samples. Methods: DNA extraction was performed using Promega Wizard襅 Genomic kits. PCR employing RV1/RV2 primers yielded 1 45-bp amplicons. Real-time PCR was performed with the same primers and SYBR Green ROX Plus mix. These techniques were used to analyze 100 peripheral blood samples from patients with clinical signs of the disease. Results...

  9. Real-time PCR in Food Science: Introduction.

    Science.gov (United States)

    Rodriguez-Lazaro, David; Hernandez, Marta

    2013-01-01

    Food safety and quality control programmes are increasingly applied throughout the production food chain in order to guarantee added value products as well as to minimize the risk of infection for the consumer. The development of real-time PCR has represented one of the most significant advances in food diagnostics as it provides rapid, reliable and quantitative results. These aspects become increasingly important for the agricultural and food industry. Different strategies for real-time PCR diagnostics have been developed including unspecific detection independent of the target sequence using fluorescent dyes such as SYBR Green, or by sequence-specific fluorescent oligonucleotide probes such as TaqMan probes or molecular beacons.

  10. SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells.

    OpenAIRE

    Kiltie, A E; Ryan, A J

    1997-01-01

    Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This meth...

  11. Real-time PCR detection and quantification of fish probiotic Phaeobacter strain 27-4 and fish pathogenic Vibrio in microalgae, rotifer, Artemia and first feeding turbot (Psetta maxima) larvae

    DEFF Research Database (Denmark)

    Prol, M.J.; Bruhn, Jesper Bartholin; Pintado, J.

    2009-01-01

    To develop a SYBR Green quantitative real-time PCR protocol enabling detection and quantification of a fish probiotic and two turbot pathogenic Vibrio spp. in microcosms. Phaeobacter 27-4, Vibrio anguillarum 90-11-287 and Vibrio splendidus DMC-1 were quantified as pure and mixed cultures and in p...

  12. Cross-platform comparison of SYBR® Green real-time PCR with TaqMan PCR, microarrays and other gene expression measurement technologies evaluated in the MicroArray Quality Control (MAQC study

    Directory of Open Access Journals (Sweden)

    Dial Stacey L

    2008-07-01

    Full Text Available Abstract Background The MicroArray Quality Control (MAQC project evaluated the inter- and intra-platform reproducibility of seven microarray platforms and three quantitative gene expression assays in profiling the expression of two commercially available Reference RNA samples (Nat Biotechnol 24:1115-22, 2006. The tested microarrays were the platforms from Affymetrix, Agilent Technologies, Applied Biosystems, GE Healthcare, Illumina, Eppendorf and the National Cancer Institute, and quantitative gene expression assays included TaqMan® Gene Expression PCR Assay, Standardized (Sta RT-PCR™ and QuantiGene®. The data showed great consistency in gene expression measurements across different microarray platforms, different technologies and test sites. However, SYBR® Green real-time PCR, another common technique utilized by half of all real-time PCR users for gene expression measurement, was not addressed in the MAQC study. In the present study, we compared the performance of SYBR Green PCR with TaqMan PCR, microarrays and other quantitative technologies using the same two Reference RNA samples as the MAQC project. We assessed SYBR Green real-time PCR using commercially available RT2 Profiler™ PCR Arrays from SuperArray, containing primer pairs that have been experimentally validated to ensure gene-specificity and high amplification efficiency. Results The SYBR Green PCR Arrays exhibit good reproducibility among different users, PCR instruments and test sites. In addition, the SYBR Green PCR Arrays have the highest concordance with TaqMan PCR, and a high level of concordance with other quantitative methods and microarrays that were evaluated in this study in terms of fold-change correlation and overlap of lists of differentially expressed genes. Conclusion These data demonstrate that SYBR Green real-time PCR delivers highly comparable results in gene expression measurement with TaqMan PCR and other high-density microarrays.

  13. SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells.

    Science.gov (United States)

    Kiltie, A E; Ryan, A J

    1997-07-15

    Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.

  14. Comparison of nine different real-time PCR chemistries for qualitative and quantitative applications in GMO detection.

    Science.gov (United States)

    Buh Gasparic, Meti; Tengs, Torstein; La Paz, Jose Luis; Holst-Jensen, Arne; Pla, Maria; Esteve, Teresa; Zel, Jana; Gruden, Kristina

    2010-03-01

    Several techniques have been developed for detection and quantification of genetically modified organisms, but quantitative real-time PCR is by far the most popular approach. Among the most commonly used real-time PCR chemistries are TaqMan probes and SYBR green, but many other detection chemistries have also been developed. Because their performance has never been compared systematically, here we present an extensive evaluation of some promising chemistries: sequence-unspecific DNA labeling dyes (SYBR green), primer-based technologies (AmpliFluor, Plexor, Lux primers), and techniques involving double-labeled probes, comprising hybridization (molecular beacon) and hydrolysis (TaqMan, CPT, LNA, and MGB) probes, based on recently published experimental data. For each of the detection chemistries assays were included targeting selected loci. Real-time PCR chemistries were subsequently compared for their efficiency in PCR amplification and limits of detection and quantification. The overall applicability of the chemistries was evaluated, adding practicability and cost issues to the performance characteristics. None of the chemistries seemed to be significantly better than any other, but certain features favor LNA and MGB technology as good alternatives to TaqMan in quantification assays. SYBR green and molecular beacon assays can perform equally well but may need more optimization prior to use.

  15. A Highly Sensitive and Selective Competition Assay for the Detection of Cysteine Using Mercury-Specific DNA, Hg2+ and Sybr Green I

    Directory of Open Access Journals (Sweden)

    Gang Liu

    2011-10-01

    Full Text Available We here report a rapid, sensitive, selective and label-free fluorescence detection method for cysteine (Cys. The conformation of mercury-specific DNA (MSD changes from a random coil form to a hairpin structure in the presence of Hg2+ due to the formation of a thymine-Hg2+-thymine (T-Hg2+-T complex. Cys can selectively coordinate with Hg2+ and extract it from the thymine-Hg2+-thymine complex. The hairpin structure dehybridizes and the fluorescence intensity of Sybr Green I (SG decreases upon addition of Cys because SG efficiently discriminates mercury-specific DNA and mercury-specific DNA/Hg2+ complex. The detection can be finished within 5 min with high sensitivity and selectivity. In addition, we can obtain variable dynamic ranges for Cys by changing the concentration of MSD/Hg2+.

  16. Modulation of the Singlet Oxygen Generation from the Double Strand DNA-SYBR Green I Complex Mediated by T-Melamine-T Mismatch for Visual Detection of Melamine.

    Science.gov (United States)

    Hu, Hao; Zhang, Jinyi; Ding, Yu; Zhang, Xinfeng; Xu, Kailai; Hou, Xiandeng; Wu, Peng

    2017-05-02

    Singlet oxygen ( 1 O 2 ), generated via photosensitization, has been proved to oxidize chromogenic substrates with neither H 2 O 2 oxidation nor enzyme (horseradish peroxidase, HRP) catalysis. Of the various methods for modulation of the 1 O 2 generation, DNA-controlled photosensitization received great attention. Therefore, integration of the formation/deformation DNA structures with DNA-controlled photosensitization will be extremely appealing in visual biosensor developments. Here, the stable melamine-thymine complex was explored in combination with DNA-controlled photosensitization for visual detection of melamine. A T-rich single stand DNA was utilized as the recognition unit. Upon the formation of the T-M-T complex, double stand DNA was formed, which was ready for the binding of SYBR Green I and activated the photosensitization. Subsequent oxidation of TMB allowed visual detection of melamine in dairy products, with spike-recoveries ranging from 94% to 106%.

  17. Application of reverse transcription-PCR and real-time PCR in nanotoxicity research.

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2012-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq(®) DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix.

  18. Real Science, Real Classrooms

    Science.gov (United States)

    Parker, Becky

    2017-01-01

    The Institute for Research in Schools (IRIS) was set up to give science students the opportunity to work on cutting-edge research projects. Teachers and students can access data and equipment that enable them to tackle real-life issues. As a result, not only do students gain in confidence and experience in their scientific work, they are also able…

  19. Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction.

    Science.gov (United States)

    Petersen, M I; Alvarez, I; Trono, K G; Jaworski, J P

    2018-04-11

    The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  20. Development of a Real-time PCR test for porcine group A rotavirus diagnosis

    Directory of Open Access Journals (Sweden)

    Elizabeth C.M. Marconi

    2015-01-01

    Full Text Available Group A Rotavirus (RVA is one of the most common causes of diarrhea in humans and several animal species. A SYBR-Green Real-Time polymerase chain reaction (PCR was developed to diagnose RVA from porcine fecal samples, targeting amplification of a 137-bp fragment of nonstructural protein 5 (NSP5 gene using mRNA of bovine NADH-desidrogenase-5 as exogenous internal control. Sixty-five samples were tested (25 tested positive for conventional PCR and genetic sequencing. The overall agreement (kappa was 0.843, indicating 'very good' concordance between tests, presenting 100% of relative sensitivity (25+ Real Time PCR/25+ Conventional PCR and 87.5% of relative sensitivity (35- Real Time PCR/40- Conventional PCR. The results also demonstrated high intra- and inter-assay reproducibility (coefficient of variation ≤1.42%; thus, this method proved to be a fast and sensitive approach for the diagnosis of RVA in pigs.

  1. Get real!

    DEFF Research Database (Denmark)

    Hyldgaard, Kirsten

    2017-01-01

    of the Lacanian concepts of the Symbolic, the Imaginary, and the Real. Particularly the concept of the Real is central. The Real has to be distinguished from any concept of reality. The concept of the Real refers to a pure negativity: to what cannot be represented; to what breaks down the safe opposition between...

  2. Molecular Analyses of Transgenic Plants.

    Science.gov (United States)

    Trijatmiko, Kurniawan Rudi; Arines, Felichi Mae; Oliva, Norman; Slamet-Loedin, Inez Hortense; Kohli, Ajay

    2016-01-01

    One of the major challenges in plant molecular biology is to generate transgenic plants that express transgenes stably over generations. Here, we describe some routine methods to study transgene locus structure and to analyze transgene expression in plants: Southern hybridization using DIG chemiluminescent technology for characterization of transgenic locus, SYBR Green-based real-time RT-PCR to measure transgene transcript level, and protein immunoblot analysis to evaluate accumulation and stability of transgenic protein product in the target tissue.

  3. Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose

    Czech Academy of Sciences Publication Activity Database

    Utekal, Pavol; Kocanda, Lukáš; Matoušek, P.; Wagner, P.; Bugajev, Viktor; Dráber, Petr

    2015-01-01

    Roč. 416, January (2015), s. 178-182 ISSN 0022-1759 R&D Projects: GA ČR(CZ) GBP302/12/G101; GA MPO FR-TI3/067; GA ČR(CZ) GA14-09807S; GA ČR(CZ) GA14-00703S Institutional support: RVO:68378050 Keywords : 1,2-Propanediol * real-time PCR * SYBR Green I * Taq DNA polymerase * trehalose * Unseparated blood Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.858, year: 2015

  4. Progesterone receptor isoform analysis by quantitative real-time polymerase chain reaction in formalin-fixed, paraffin-embedded canine mammary dysplasias and tumors

    DEFF Research Database (Denmark)

    Guil-Luna, S.; Stenvang, Jan; Brünner, Nils

    2014-01-01

    and its isoforms in formalin-fixed, paraffin-embedded tissue samples from canine mammary lesions (4 dysplasias, 10 benign tumors, and 46 carcinomas) using 1-step SYBR Green quantitative real-time polymerase chain reaction (RT-qPCR). Progesterone receptor was expressed in 75% of dysplasias, all benign...... in the expression of isoform A versus B. Analysis of progesterone receptor mRNA isoforms by RT-qPCR was successful in routinely formalin-fixed, paraffin-embedded tissue samples and enabled the distribution of isoforms A and B to be identified for the first time in dysplasias, benign tumors, and malignant tumors...

  5. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

    Directory of Open Access Journals (Sweden)

    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  6. Real Language Meets Real Business

    Science.gov (United States)

    Muirhead, Muirhead; Schechter, Sarah

    2016-01-01

    The Real Business Language Challenge was a collaborative pilot project between Coca-Cola Enterprises (CCE) and Routes into Languages East for Year 9 and 10 pupils. It was based on CCE's award-winning Real Business Challenge, part of its highly acclaimed education programme. The Real Business Language Challenge transformed the project into a…

  7. Real Forestry for Real Estate

    Science.gov (United States)

    Gagnon, Jennifer; Fisher, Jason

    2013-01-01

    Virginia is poised to see an unprecedented change in forest land ownership. To provide new landowners with information on sustainable forest management, we developed a two-part program, Real Forestry for Real Estate. First, we assembled New Landowner Packets, which contain a variety of sustainable forest management resources. Second, two…

  8. Development and Characterization of a Highly Specific and Sensitive SYBR Green Reverse Transcriptase PCR Assay for Detection of the 2009 Pandemic H1N1 Influenza Virus on the Basis of Sequence Signatures▿ †

    Science.gov (United States)

    Medina, Rafael A.; Rojas, Mark; Tuin, Astrid; Huff, Stephen; Ferres, Marcela; Martinez-Valdebenito, Constanza; Godoy, Paula; García-Sastre, Adolfo; Fofanov, Yuriy; SantaLucia, John

    2011-01-01

    The emergence and rapid spread of the 2009 H1N1 pandemic influenza virus showed that many diagnostic tests were unsuitable for detecting the novel virus isolates. In most countries the probe-based TaqMan assay developed by the U.S. Centers for Disease Control and Prevention was used for diagnostic purposes. The substantial sequence data that became available during the course of the pandemic created the opportunity to utilize bioinformatics tools to evaluate the unique sequence properties of this virus for the development of diagnostic tests. We used a comprehensive computational approach to examine conserved 2009 H1N1 sequence signatures that are at least 20 nucleotides long and contain at least two mismatches compared to any other known H1N1 genome. We found that the hemagglutinin (HA) and neuraminidase (NA) genes contained sequence signatures that are highly conserved among 2009 H1N1 isolates. Based on the NA gene signatures, we used Visual-OMP to design primers with optimal hybridization affinity and we used ThermoBLAST to minimize amplification artifacts. This procedure resulted in a highly sensitive and discriminatory 2009 H1N1 detection assay. Importantly, we found that the primer set can be used reliably in both a conventional TaqMan and a SYBR green reverse transcriptase (RT)-PCR assay with no loss of specificity or sensitivity. We validated the diagnostic accuracy of the NA SYBR green assay with 125 clinical specimens obtained between May and August 2009 in Chile, and we showed diagnostic efficacy comparable to the CDC assay. Our approach highlights the use of systematic computational approaches to develop robust diagnostic tests during a viral pandemic. PMID:21084522

  9. Use of 65 kDa mannoprotein gene primers in Real Time PCR identification of Candida albicans in biological samples.

    Science.gov (United States)

    Arancia, Silvia; Carattoli, Alessandra; La Valle, Roberto; Cassone, Antonio; De Bernardis, Flavia

    2006-10-01

    A method for the detection and quantification of Candida albicans in biological samples (blood, urine and serum) was developed with the use of Real-Time PCR utilizing CaMP65-specific primers. Two different systems were used for the detection in the LightCycler platform (Roche): the SYBR green fluorescent dye with melting peak analysis and the 5'nuclease fluorescent-probe detection. The amplification was highly specific for C. albicans, providing no cross-reaction on genomic DNA extracted from other Candida species or Aspergillus. The sensitivity in simulated biological samples was especially high (1 genome) when applied to sera and urine, and in blood samples the limit of detection was higher by ten-fold. Finally, the real-time PCR was employed in order to detect and quantify C. albicans in the sera from patients with invasive candidiasis.

  10. The developmental stage of strongyle eggs affects the outcome of real-time PCR analysis

    DEFF Research Database (Denmark)

    Roust, Tina; Haakansson, Ida T.; Rhod, Maria

    Several molecular diagnostic tests are based upon measuring and quantifying DNA obtained from parasite eggs. It is well-known that such eggs undergo development during storage, but it remains unknown to which extent the stage of development can affect the diagnostic test result. This project...... investigated whether the developmental stage of strongyle eggs affects real-time polymerase chain reaction (PCR) results. Mixed species strongyle eggs were obtained from the faeces of a naturally infected horse. Eggs were isolated and placed in microtiter plates with demineralised water. A total of 25 wells...... using a commercial kit previously validated for strongyle type eggs. PCR reactions were performed with a primer set specific for the ribosomal DNA region for all strongyle type parasites (NC1, NC2). PCRs were performed in triplicates using SYBR Green as fluorescent dye. PCR results were registered...

  11. Real analysis

    CERN Document Server

    McShane, Edward James

    2013-01-01

    This text surveys practical elements of real function theory, general topology, and functional analysis. Discusses the maximality principle, the notion of convergence, the Lebesgue-Stieltjes integral, function spaces and harmonic analysis. Includes exercises. 1959 edition.

  12. Real analysis

    CERN Document Server

    Loeb, Peter A

    2016-01-01

    This textbook is designed for a year-long course in real analysis taken by beginning graduate and advanced undergraduate students in mathematics and other areas such as statistics, engineering, and economics. Written by one of the leading scholars in the field, it elegantly explores the core concepts in real analysis and introduces new, accessible methods for both students and instructors. The first half of the book develops both Lebesgue measure and, with essentially no additional work for the student, general Borel measures for the real line. Notation indicates when a result holds only for Lebesgue measure. Differentiation and absolute continuity are presented using a local maximal function, resulting in an exposition that is both simpler and more general than the traditional approach. The second half deals with general measures and functional analysis, including Hilbert spaces, Fourier series, and the Riesz representation theorem for positive linear functionals on continuous functions with compact support....

  13. Real Closed Rings and Real Closed * Rings

    OpenAIRE

    Capco, Jose

    2007-01-01

    Here we try to distinguish and compare different notions of real closedness mainly one developed by N. Schwartz in his Habilitationschrift and the other developed by A. Sankaranand K. Varadarajan which we shall call real closed *. We stick to the definition of real closed rings as defined and characterized N. Schwartz and we try to determine and characterize real closed rings that are real closed *. The main result is that real closed rings have unique real closure * and that real closure of ...

  14. Real analysis

    CERN Document Server

    DiBenedetto, Emmanuele

    2016-01-01

    The second edition of this classic textbook presents a rigorous and self-contained introduction to real analysis with the goal of providing a solid foundation for future coursework and research in applied mathematics. Written in a clear and concise style, it covers all of the necessary subjects as well as those often absent from standard introductory texts. Each chapter features a “Problems and Complements” section that includes additional material that briefly expands on certain topics within the chapter and numerous exercises for practicing the key concepts. The first eight chapters explore all of the basic topics for training in real analysis, beginning with a review of countable sets before moving on to detailed discussions of measure theory, Lebesgue integration, Banach spaces, functional analysis, and weakly differentiable functions. More topical applications are discussed in the remaining chapters, such as maximal functions, functions of bounded mean oscillation, rearrangements, potential theory, a...

  15. Direct comparison of the histidine-rich protein-2 enzyme-linked immunosorbent assay (HRP-2 ELISA) and malaria SYBR green I fluorescence (MSF) drug sensitivity tests in Plasmodium falciparum reference clones and fresh ex vivo field isolates from Cambodia.

    Science.gov (United States)

    Chaorattanakawee, Suwanna; Tyner, Stuart D; Lon, Chanthap; Yingyuen, Kritsanai; Ruttvisutinunt, Wiriya; Sundrakes, Siratchana; Sai-gnam, Piyaporn; Johnson, Jacob D; Walsh, Douglas S; Saunders, David L; Lanteri, Charlotte A

    2013-07-12

    Performance of the histidine-rich protein-2 enzyme-linked immunosorbent assay (HRP-2 ELISA) and malaria SYBR Green I fluorescence (MSF) drug sensitivity tests were directly compared using Plasmodium falciparum reference strains and fresh ex vivo isolates from Cambodia against a panel of standard anti-malarials. The objective was to determine which of these two common assays is more appropriate for studying drug susceptibility of "immediate ex vivo" (IEV) isolates, analysed without culture adaption, in a region of relatively low malaria transmission. Using the HRP-2 and MSF methods, the 50% inhibitory concentration (IC50) values against a panel of malaria drugs were determined for P. falciparum reference clones (W2, D6, 3D7 and K1) and 41 IEV clinical isolates from an area of multidrug resistance in Cambodia. Comparison of the IC50 values from the two methods was made using Wilcoxon matched pair tests and Pearson's correlation. The lower limit of parasitaemia detection for both methods was determined for reference clones and IEV isolates. Since human white blood cell (WBC) DNA in clinical samples is known to reduce MSF assay sensitivity, SYBR Green I fluorescence linearity of P. falciparum samples spiked with WBCs was evaluated to assess the relative degree to which MSF sensitivity is reduced in clinical samples. IC50 values correlated well between the HRP-2 and MSF methods when testing either P. falciparum reference clones or IEV isolates against 4-aminoquinolines (chloroquine, piperaquine and quinine) and the quinoline methanol mefloquine (Pearson r = 0.85-0.99 for reference clones and 0.56-0.84 for IEV isolates), whereas a weaker IC50 value correlation between methods was noted when testing artemisinins against reference clones and lack of correlation when testing IEV isolates. The HRP-2 ELISA produced a higher overall success rate (90% for producing IC50 best-fit sigmoidal curves), relative to only a 40% success rate for the MSF assay, when evaluating ex

  16. Comparison between Conventional and Real-Time PCR Assays for Diagnosis of Visceral Leishmaniasis

    Directory of Open Access Journals (Sweden)

    Mariana R. Pereira

    2014-01-01

    Full Text Available The diagnosis of visceral leishmaniasis (VL is a challenging issue and several studies worldwide have evaluated the different tools to reach a diagnostic solution. The polymerase chain reaction (PCR has proven to be effective in detecting the genome of Leishmania species in different biological samples. In this study, we compared the conventional PCR and real-time PCR using the Sybr Green system and their application in molecular diagnosis of visceral leishmaniasis in peripheral blood as a biological sample. The genus-specific conserved region of kinetoplast DNA (kDNA was the target of amplification. We studied 30 samples from patients with suspect of visceral leishmaniasis who were treated by the Medical Clinic of Santa Casa de Belo Horizonte Hospital, Brazil. Among the samples studied, 19 had a confirmed diagnosis for VL by serology and/or by clinical findings. Among these 19 samples, 63% (n=12 presented positive results for serology and 79% (n=15 positive results in both PCR methodologies. This fact suggests that the PCR technique can assist in the diagnosis of visceral leishmaniasis in patients who do not have detectable antibodies by serology but can present the genome of the parasite circulating in whole blood. Also, it was possible to observe that there was conformity between the results of the techniques of cPCR and qPCR using the Sybr Green system in 100% of samples analyzed. These data suggest that both PCR techniques were equally effective for detection of the genome of the parasite in the patient’s blood.

  17. An Improved Multiplex Real-Time SYBR Green PCR Assay for Analysis of 24 Target Genes from 16 Bacterial Species in Fecal DNA Samples from Patients with Foodborne Illnesses.

    Science.gov (United States)

    Kawase, Jun; Etoh, Yoshiki; Ikeda, Tetsuya; Yamaguchi, Keiji; Watahiki, Masanori; Shima, Tomoko; Kameyama, Mitsuhiro; Horikawa, Kazumi; Fukushima, Hiroshi; Goto, Ryoichi; Shirabe, Komei

    2016-05-20

    Here, we developed a new version of our original screening system (Rapid Foodborne Bacterial Screening 24; RFBS24), which can simultaneously detect 24 genes of foodborne pathogens in fecal DNA samples. This new version (RFBS24 ver. 5) detected all known stx2 subtypes, enterotoxigenic Escherichia coli (STh genotype), and Vibrio parahaemolyticus (trh2), which were not detected by the original RFBS24 assay. The detection limits of RFBS24 ver. 5 were approximately 5.6 × 10(-2)-5.6 × 10(-5) (ng DNA)/reaction, significantly lower (10- to 100-fold) than those of the original RFBS24 for the 22 target genes analyzed here. We also tested the new assay on fecal DNA samples from patients infected with Salmonella, Campylobacter, or enterohemorrhagic E. coli. The number of bacterial target genes detected by RFBS24 ver. 5 was greater than that detected by RFBS24. RFBS24 ver. 5 combined with an Ultra Clean Fecal DNA Isolation Kit showed adequate performance (sensitivity and specificity 89% and 100%, respectively, for Salmonella spp. and 100% and 83%, respectively, for Campylobacter jejuni) in terms of rapid detection of a causative pathogen during foodborne-illness outbreaks. Thus, RFBS24 ver. 5 is more useful than the previous assay system for detection of foodborne pathogens and offers quick simultaneous analysis of many targets and thus facilitates rapid dissemination of information to public health officials.

  18. Direct quantification of fungal DNA from soil substrate using real-time PCR.

    Science.gov (United States)

    Filion, Martin; St-Arnaud, Marc; Jabaji-Hare, Suha H

    2003-04-01

    Detection and quantification of genomic DNA from two ecologically different fungi, the plant pathogen Fusarium solani f. sp. phaseoli and the arbuscular mycorrhizal fungus Glomus intraradices, was achieved from soil substrate. Specific primers targeting a 362-bp fragment from the SSU rRNA gene region of G. intraradices and a 562-bp fragment from the F. solani f. sp. phaseoli translation elongation factor 1 alpha gene were used in real-time polymerase chain reaction (PCR) assays conjugated with the fluorescent SYBR(R) Green I dye. Standard curves showed a linear relation (r(2)=0.999) between log values of fungal genomic DNA of each species and real-time PCR threshold cycles and were quantitative over 4-5 orders of magnitude. Real-time PCR assays were applied to in vitro-produced fungal structures and sterile and non-sterile soil substrate seeded with known propagule numbers of either fungi. Detection and genomic DNA quantification was obtained from the different treatments, while no amplicon was detected from non-seeded non-sterile soil samples, confirming the absence of cross-reactivity with the soil microflora DNA. A significant correlation (Pgenomic DNA of F. solani f. sp. phaseoli or G. intraradices detected and the number of fungal propagules present in seeded soil substrate. The DNA extraction protocol and real-time PCR quantification assay can be performed in less than 2 h and is adaptable to detect and quantify genomic DNA from other soilborne fungi.

  19. Quantitative detection of residual E. coli host cell DNA by real-time PCR.

    Science.gov (United States)

    Lee, Dong Hyuck; Bae, Jung Eun; Lee, Jung Hee; Shin, Jeong Sup; Kim, In Seop

    2010-10-01

    E. coli has been widely used as a host system to manufacture recombinant proteins for human therapeutic use. Among impurities to be eliminated during the downstream process, residual host cell DNA is a major interest for safety. Residual E. coli host cell DNA in the final products are usually determined using conventional slot blot hybridization assay or total DNA Threshold assay, although these methods are time consuming, expensive, and relatively insensitive. Therefore a sensitive real-time PCR assay for specific detection of residual E. coli DNA was developed and compared with slot blot hybridization assay and Threshold assay to validate the overall capability of these methods. Specific primer pair for amplification of the E. coli 16S rRNA gene was selected to improve the sensitivity, and E. coli host cell DNA was quantified by use of SYBR Green 1. The detection limit of real-time PCR assay in the optimized condition was calculated to be 0.042 pg genomic DNA, which is much higher than those of slot blot hybridization assay and Threshold assay of which detection limit were 2.42 and 3.73 pg genomic DNA, respectively. The real-time PCR assay was validated to be more reproducible, accurate, and precise than slot blot hybridization assay and Threshold assay. The real-time PCR assay may be a useful tool for quantitative detection and clearance validation of residual E. coli DNA during the manufacturing process for recombinant therapeutics.

  20. REAL-TIME PCR DETECTION OF LISTERIA MONOCYTOGENES IN FOOD SAMPLES OF ANIMAL ORIGIN

    Directory of Open Access Journals (Sweden)

    Jaroslav Pochop

    2013-02-01

    Full Text Available The aim of this study was to follow the contamination of food with Listeria monocytogenes by using Step One real time polymerase chain reaction (PCR. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In 24 samples of food of animal origin without incubation were detected strains of Listeria monocytogenes in 15 samples (swabs. Nine samples were negative. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in food of animal origin without incubation. This could prevent infection caused by Listeria monocytogenes, and also could benefit food manufacturing companies by extending their product’s shelf-life as well as saving the cost of warehousing their food products while awaiting pathogen testing results. The rapid real-time PCR-based method performed very well compared to the conventional method. It is a fast, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future.

  1. Comentarios reales

    OpenAIRE

    Brusa, Orietta

    2014-01-01

    El término real nada tiene que ver con la realeza, pero sí mucho con la realidad. De hecho, difícilmente los que manejan el poder (reyes, presidentes, empresarios, papas y/o cardenales, autoridades en general) tienen un concepto adecuado de la realidad. Es como si al lado de las dimensiones paralelas existieran realidades paralelas que, por supuesto, nunca se encuentran. Será porque ellos y los comunes mortales no viven en el mismo plan existencial.

  2. Development of a high-throughput real time PCR based on a hot-start alternative for Pfu mediated by quantum dots

    Science.gov (United States)

    Sang, Fuming; Yang, Yang; Yuan, Lin; Ren, Jicun; Zhang, Zhizhou

    2015-09-01

    Hot start (HS) PCR is an excellent alternative for high-throughput real time PCR due to its ability to prevent nonspecific amplification at low temperature. Development of a cost-effective and simple HS PCR technique to guarantee high-throughput PCR specificity and consistency still remains a great challenge. In this study, we systematically investigated the HS characteristics of QDs triggered in real time PCR with EvaGreen and SYBR Green I dyes by the analysis of amplification curves, standard curves and melting curves. Two different kinds of DNA polymerases, Pfu and Taq, were employed. Here we showed that high specificity and efficiency of real time PCR were obtained in a plasmid DNA and an error-prone two-round PCR assay using QD-based HS PCR, even after an hour preincubation at 50 °C before real time PCR. Moreover, the results obtained by QD-based HS PCR were comparable to a commercial Taq antibody DNA polymerase. However, no obvious HS effect of QDs was found in real time PCR using Taq DNA polymerase. The findings of this study demonstrated that a cost-effective high-throughput real time PCR based on QD triggered HS PCR could be established with high consistency, sensitivity and accuracy.Hot start (HS) PCR is an excellent alternative for high-throughput real time PCR due to its ability to prevent nonspecific amplification at low temperature. Development of a cost-effective and simple HS PCR technique to guarantee high-throughput PCR specificity and consistency still remains a great challenge. In this study, we systematically investigated the HS characteristics of QDs triggered in real time PCR with EvaGreen and SYBR Green I dyes by the analysis of amplification curves, standard curves and melting curves. Two different kinds of DNA polymerases, Pfu and Taq, were employed. Here we showed that high specificity and efficiency of real time PCR were obtained in a plasmid DNA and an error-prone two-round PCR assay using QD-based HS PCR, even after an hour

  3. Development of a real time polymerase chain reaction for quantitation of Schistosoma mansoni DNA

    Directory of Open Access Journals (Sweden)

    Ana Lisa do Vale Gomes

    2006-10-01

    Full Text Available This report describes the development of a SYBR Green I based real time polymerase chain reaction (PCR protocol for detection on the ABI Prism 7000 instrument. Primers targeting the gene encoding the SSU rRNA were designed to amplify with high specificity DNA from Schistosoma mansoni, in a real time quantitative PCR system. The limit of detection of parasite DNA for the system was 10 fg of purified genomic DNA, that means less than the equivalent to one parasite cell (genome ~580 fg DNA. The efficiency was 0.99 and the correlation coefficient (R² was 0.97. When different copy numbers of the target amplicon were used as standards, the assay could detect at least 10 copies of the specific target. The primers used were designed to amplify a 106 bp DNA fragment (Tm 83ºC. The assay was highly specific for S. mansoni, and did not recognize DNA from closely related non-schistosome trematodes. The real time PCR allowed for accurate quantification of S. mansoni DNA and no time-consuming post-PCR detection of amplification products by gel electrophoresis was required. The assay is potentially able to quantify S. mansoni DNA (and indirectly parasite burden in a number of samples, such as snail tissue, serum and feces from patients, and cercaria infested water. Thus, these PCR protocols have potential to be used as tools for monitoring of schistosome transmission and quantitative diagnosis of human infection.

  4. A Highly Sensitive and Selective Method for the Determination of an Iodate in Table-salt Samples Using Malachite Green-based Spectrophotometry.

    Science.gov (United States)

    Konkayan, Mongkol; Limchoowong, Nunticha; Sricharoen, Phitchan; Chanthai, Saksit

    2016-01-01

    A simple, rapid, and sensitive malachite green-based spectrophotometric method for the selective trace determination of an iodate has been developed and presented for the first time. The reaction mixture was specifically involved in the liberation of iodine in the presence of an excess of iodide in an acidic condition following an instantaneous reaction between the liberated iodine and malachite green dye. The optimum condition was obtained with a buffer solution pH of 5.2 in the presence of 40 mg L -1 potassium iodide and 1.5 × 10 -5 M malachite green for a 5-min incubation time. The iodate contents in some table-salt samples were in the range of 26 to 45 mg kg -1 , while those of drinking water, tap water, canal water, and seawater samples were not detectable (< 96 ng mL -1 of limits of detection, LOQ) with their satisfied method of recoveries of between 93 and 108%. The results agreed with those obtained using ICP-OES for comparison.

  5. Detection of Campylobacter jejuni in naturally contaminated chicken skin by melting peak analysis of amplicons in real-time PCR.

    Science.gov (United States)

    Oliveira, Tereza C R M; Barbut, Shai; Griffiths, Mansel W

    2005-09-25

    Contamination of poultry by Campylobacter spp. is a significant source of human diarrheal diseases. Traditional methods currently used to detect Campylobacter in foods are time-consuming and labor-intensive. In this study, primers designed for the Campylobacter jejuni cadF gene sequence were used in a SYBR Green I real-time PCR assay as an alternative to a conventional bacteriological method for the rapid detection of C. jejuni from poultry. Twelve portions of chicken purchased from two local grocery stores and 39 portions obtained from a commercial processing plant were examined. Samples of the skin were enriched in Bolton broth at 37 degrees C for 3 h and then at 42 degrees C for 9, 21, or 45 h under microaerobic conditions. DNA was extracted from 1-ml aliquots of the enrichment cultures using 1% Triton X-100. The DNA was used as the template in a real-time polymerase chain reaction (PCR) assay. After 24 h of enrichment, C. jejuni was isolated from 13 samples and all of the positive cultures were also detected by the real-time PCR procedure. C. jejuni was detected by both methods from samples artificially contaminated with 1 or 10 CFU of C. jejuni per 10 g, after 24 h of enrichment. The real-time PCR method was found to be sensitive and specific. It significantly reduced the time required for the detection of C. jejuni in poultry following enrichment of samples.

  6. Novel real-time PCr for detection of Schistosoma japonicum in stool.

    Science.gov (United States)

    Lier, T; Simonsen, G S; Haaheim, H; Hjelmevoll, S O; Vennervald, B J; Johansen, M V

    2006-03-01

    Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

  7. Optimization of human cytomegalovirus LightCycler real-time PCR.

    Science.gov (United States)

    Habbal, Wafa; Monem, Fawza; Gärtner, Barbara Christine

    2008-10-01

    Real-time PCR has been widely considered as a powerful tool for the evaluation of Human Cytomegalovirus (CMV) DNA kinetics. Successful PCR relies on optimization, which is an extremely demanding procedure. Nevertheless, certain values could be optimal for most primers in use. Seventeen CMV primer sets recommended in the literature were selected for optimization in terms of MgCl2 and primers concentrations as well as annealing temperature using the LightCycler instrument and SYBR Green I detection format. Optimal values were considered as those showing the lowest crossing point (Cp), the highest fluorescence intensity, the steepest sigmoid curve slope, and the absence of non-specific PCR products. Optimal values for most studied primers were found to be 3 mM for MgCl2 concentration, 0.5 microM and 0.6 microM for primers concentration, and 55 degrees C for annealing temperature. Adopting the resulting values for CMV-specific primers generally used in single-target real-time PCR assays with the same thermal cycler may guarantee their efficient performance minimizing cost and time needed for optimization.

  8. Single reaction, real time RT-PCR detection of all known avian and human metapneumoviruses.

    Science.gov (United States)

    Lemaitre, E; Allée, C; Vabret, A; Eterradossi, N; Brown, P A

    2018-01-01

    Current molecular methods for the detection of avian and human metapneumovirus (AMPV, HMPV) are specifically targeted towards each virus species or individual subgroups of these. Here a broad range SYBR Green I real time RT-PCR was developed which amplified a highly conserved fragment of sequence in the N open reading frame. This method was sufficiently efficient and specific in detecting all MPVs. Its validation according to the NF U47-600 norm for the four AMPV subgroups estimated low limits of detection between 1000 and 10copies/μL, similar with detection levels described previously for real time RT-PCRs targeting specific subgroups. RNA viruses present a challenge for the design of durable molecular diagnostic test due to the rate of change in their genome sequences which can vary substantially in different areas and over time. The fact that the regions of sequence for primer hybridization in the described method have remained sufficiently conserved since the AMPV and HMPV diverged, should give the best chance of continued detection of current subgroups and of potential unknown or future emerging MPV strains. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Real-time detection of an airborne microorganism using inertial impaction and mini-fluorescent microscopy.

    Science.gov (United States)

    Kang, Joon Sang; Lee, Kang Soo; Kim, Sang Soo; Bae, Gwi-Nam; Jung, Jae Hee

    2014-01-07

    To achieve successful real-time detection of airborne pathogenic microorganisms, the problem must be considered in terms of their physical size and biological characteristics. We developed an airborne microorganism detection chip to realize the detection of microorganisms, ensuring compactness, sensitivity, cost-efficiency, and portability, using three key components: an inertial impaction system, a cartridge-type impaction plate, and a mini-fluorescent microscope. The inertial impaction system was used to separate microorganisms in terms of their aerodynamic particle size, and was fabricated with three impaction stages. Numerical analysis was performed to design the system; the calculated cutoff diameter at each impaction stage was 2.02 (first stage), 0.88 (second stage), and 0.54 μm (third stage). The measured cutoff diameters were 2.24, 0.91, and 0.49 μm, respectively. A cartridge-type impaction plate was used, composed of molded polydimethylsiloxane (PDMS) and an actual impaction region made of a SYBR green I dye-stained agar plate. A mini-fluorescent microscope was used to distinguish microbes from non-biological particles. Images of the microorganisms deposited at the impaction zone were obtained via mini-fluorescent microscopy, and fluorescent intensities of the images were calculated using in-house image-processing software. The results showed that the developed system successfully identified aerosolized biological particles from non-biological particles in real time.

  10. Real-Time Polymerase Chain Reaction Quantification of Phytophthora capsici in Different Pepper Genotypes.

    Science.gov (United States)

    Silvar, C; Díaz, J; Merino, F

    2005-12-01

    ABSTRACT Reliable and sensitive quantification of Phytophthora capsici in pepper plants is of crucial importance in managing the multiple syndromes caused by this pathogen. A real-time polymerase chain reaction (PCR) assay was developed for the determination of P. capsici in pepper tissues. DNA levels of a highly virulent and a less virulent isolate were measured in different pepper genotypes with varying degrees of resistance. Using SYBR Green and specific primers for P. capsici, the minimal amount of pathogen DNA quantified was 10 pg. Pathogen DNA was recorded as early as 8 h postinoculation. Thereafter, the increase was rapid in susceptible cultivars and slower in resistant ones. The amount of pathogen DNA quantified in each pepper genotype correlated with susceptibility to Phytophthora root rot. Likewise, there was a relationship between the virulence of the pathogen and the degree of colonization. Differences also were found in oomycete amount among pepper tissues, with maximal pathogen biomass occurring in stems. The real-time PCR technique developed in this study was sensitive and robust enough to assess both pathogen development and resistance to Phytophthora root rot in different pepper genotypes.

  11. Real-time RT-PCR profiling of over 1400 Arabidopsis transcription factors: unprecedented sensitivity reveals novel root- and shoot-specific genes.

    Science.gov (United States)

    Czechowski, Tomasz; Bari, Rajendra P; Stitt, Mark; Scheible, Wolf-Rüdiger; Udvardi, Michael K

    2004-04-01

    Summary To overcome the detection limits inherent to DNA array-based methods of transcriptome analysis, we developed a real-time reverse transcription (RT)-PCR-based resource for quantitative measurement of transcripts for 1465 Arabidopsis transcription factors (TFs). Using closely spaced gene-specific primer pairs and SYBR Green to monitor amplification of double-stranded DNA (dsDNA), transcript levels of 83% of all target genes could be measured in roots or shoots of young Arabidopsis wild-type plants. Only 4% of reactions produced non-specific PCR products. The amplification efficiency of each PCR was determined from the log slope of SYBR Green fluorescence versus cycle number in the exponential phase, and was used to correct the readout for each primer pair and run. Measurements of transcript abundance were quantitative over six orders of magnitude, with a detection limit equivalent to one transcript molecule in 1000 cells. Transcript levels for different TF genes ranged between 0.001 and 100 copies per cell. Only 13% of TF transcripts were undetectable in these organs. For comparison, 22K Arabidopsis Affymetrix chips detected less than 55% of TF transcripts in the same samples, the range of transcript levels was compressed by a factor more than 100, and the data were less accurate especially in the lower part of the response range. Real-time RT-PCR revealed 35 root-specific and 52 shoot-specific TF genes, most of which have not been identified as organ-specific previously. Finally, many of the TF transcripts detected by RT-PCR are not represented in Arabidopsis EST (expressed sequence tag) or Massively Parallel Signature Sequencing (MPSS) databases. These genes can now be annotated as expressed.

  12. A one-step real-time multiplex PCR for screening Y-chromosomal microdeletions without downstream amplicon size analysis.

    Directory of Open Access Journals (Sweden)

    Viviana Kozina

    Full Text Available BACKGROUND: Y-chromosomal microdeletions (YCMD are one of the major genetic causes for non-obstructive azoospermia. Genetic testing for YCMD by multiplex polymerase chain reaction (PCR is an established method for quick and robust screening of deletions in the AZF regions of the Y-chromosome. Multiplex PCRs have the advantage of including a control gene in every reaction and significantly reducing the number of reactions needed to screen the relevant genomic markers. PRINCIPAL FINDINGS: The widely established "EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions (2004" were used as a basis for designing a real-time multiplex PCR system, in which the YCMD can simply be identified by their melting points. For this reason, some AZF primers were substituted by primers for regions in their genomic proximity, and the ZFX/ZFY control primer was exchanged by the AMELX/AMELY control primer. Furthermore, we substituted the classical SybrGreen I dye by the novel and high-performing DNA-binding dye EvaGreen™ and put substantial effort in titrating the primer combinations in respect to optimal melting peak separation and peak size. SIGNIFICANCE: With these changes, we were able to develop a platform-independent and robust real-time based multiplex PCR, which makes the need for amplicon identification by electrophoretic sizing expendable. By using an open-source system for real-time PCR analysis, we further demonstrate the applicability of automated melting point and YCMD detection.

  13. A sensitive, reproducible, and economic real-time reverse transcription PCR detecting avian metapneumovirus subtypes A and B.

    Science.gov (United States)

    Franzo, G; Drigo, M; Lupini, C; Catelli, E; Laconi, A; Listorti, V; Bonci, M; Naylor, C J; Martini, M; Cecchinato, M

    2014-06-01

    Use of real-time PCR is increasing in the diagnosis of infectious disease due to its sensitivity, specificity, and speed of detection. These characteristics make it particularly suited for the diagnosis of viral infections, like avian metapneumovirus (AMPV), for which effective control benefits from continuously updated knowledge of the epidemiological situation. Other real-time reverse transcription (RT)-PCRs have been published based on highly specific fluorescent dye-labeled probes, but they have high initial cost, complex validation, and a marked susceptibility to the genetic variability of their target sequence. With this in mind, we developed and validated a SYBR Green I-based quantitative RT-PCR for the detection of the two most prevalent AMPV subtypes (i.e., subtypes A and B). The assay demonstrated an analytical sensitivity comparable with that of a previously published real-time RT-PCR and the ability to detect RNA equivalent to approximately 0.5 infectious doses for both A and B subtypes. The high efficiency and linearity between viral titer and crossing point displayed for both subtypes make it suited for viral quantification. Optimization of reaction conditions and the implementation of melting curve analysis guaranteed the high specificity of the assay. The stable melting temperature difference between the two subtypes indicated the possibility of subtyping through melting temperature analysis. These characteristics make our assay a sensitive, specific, and rapid tool, enabling contemporaneous detection, quantification, and discrimination of AMPV subtype A and B.

  14. Quantitative detection of Cucumber vein yellowing virus in susceptible and partially resistant plants using real-time PCR.

    Science.gov (United States)

    Picó, Belén; Sifres, Alicia; Nuez, Fernando

    2005-09-01

    A method for the detection of Cucumber vein yellowing virus (CVYV) that combines reverse transcription with real-time PCR (SYBR((R)) Green chemistry) was developed using specific primers designed from a nucleotide sequence of the RNA polymerase gene (NIb) conserved among all the available CVYV strains. This method provided a linear assay over five to six orders of magnitude and reproducibly detected titres as low as 10(3) molecules of the target CVYV cDNA. Real-time PCR gave reproducible results for the quantification of CVYV in young leaves of susceptible and resistant cucumber landraces after mechanical inoculation. Significant differences in the starting amount of target cDNA were found between the analyzed genotypes, indicating differences in viral accumulation that correlated to their different levels of resistance. Real-time PCR results validated our previous findings using slot-blot hybridization, the dominance of the strong resistance to CVYV displayed by C.sat 10, and provided improved reliability and sensitivity of detection. This method has great potential in resistance breeding for germplasm screening, characterization of resistance mechanisms and genetic studies.

  15. Real Algebraic Geometry

    CERN Document Server

    Mahé, Louis; Roy, Marie-Françoise

    1992-01-01

    Ten years after the first Rennes international meeting on real algebraic geometry, the second one looked at the developments in the subject during the intervening decade - see the 6 survey papers listed below. Further contributions from the participants on recent research covered real algebra and geometry, topology of real algebraic varieties and 16thHilbert problem, classical algebraic geometry, techniques in real algebraic geometry, algorithms in real algebraic geometry, semialgebraic geometry, real analytic geometry. CONTENTS: Survey papers: M. Knebusch: Semialgebraic topology in the last ten years.- R. Parimala: Algebraic and topological invariants of real algebraic varieties.- Polotovskii, G.M.: On the classification of decomposing plane algebraic curves.- Scheiderer, C.: Real algebra and its applications to geometry in the last ten years: some major developments and results.- Shustin, E.L.: Topology of real plane algebraic curves.- Silhol, R.: Moduli problems in real algebraic geometry. Further contribu...

  16. Real-time PCR (qPCR) primer design using free online software.

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most commonly used fluorescent chemistries are SYBR® Green dyes and TaqMan®, Molecular Beacon or Scorpion probes. SYBR® Green is very simple to use and cost efficient. As SYBR® Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR® Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR® Green-based primers with free, online software. Copyright © 2010 Wiley Periodicals, Inc.

  17. Real-Time PCR Quantification of Chloroplast DNA Supports DNA Barcoding of Plant Species.

    Science.gov (United States)

    Kikkawa, Hitomi S; Tsuge, Kouichiro; Sugita, Ritsuko

    2016-03-01

    Species identification from extracted DNA is sometimes needed for botanical samples. DNA quantification is required for an accurate and effective examination. If a quantitative assay provides unreliable estimates, a higher quantity of DNA than the estimated amount may be used in additional analyses to avoid failure to analyze samples from which extracting DNA is difficult. Compared with conventional methods, real-time quantitative PCR (qPCR) requires a low amount of DNA and enables quantification of dilute DNA solutions accurately. The aim of this study was to develop a qPCR assay for quantification of chloroplast DNA from taxonomically diverse plant species. An absolute quantification method was developed using primers targeting the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene using SYBR Green I-based qPCR. The calibration curve was generated using the PCR amplicon as the template. DNA extracts from representatives of 13 plant families common in Japan. This demonstrates that qPCR analysis is an effective method for quantification of DNA from plant samples. The results of qPCR assist in the decision-making will determine the success or failure of DNA analysis, indicating the possibility of optimization of the procedure for downstream reactions.

  18. Real-time PCR for quantitative detection of chamois (Rupicapra rupicapra) and pyrenean ibex (Capra pyrenaica) in meat mixtures.

    Science.gov (United States)

    Fajardo, Violeta; González, Isabel; Martín, Irene; Rojas, María; Hernández, Pablo E; García, Teresa; Martín, Rosario

    2008-01-01

    A real-time quantitative polymerase chain reaction (PCR) technique was developed for the quantification of chamois and pyrenean ibex DNAs in meat mixtures by using a SYBR green detection platform. Two species-specific systems and a eukaryotic endogenous system were combined in the real-time PCR approach to quantify the target species. In the specific systems, a 133 base pair (bp) fragment of the 12S rRNA gene was amplified from chamois DNA, and an 88 bp fragment from the D-loop region was amplified from pyrenean ibex DNA. In the endogenous system, universal primers amplified a 141 bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. The threshold cycle values obtained with the 18S rRNA primers were used to normalize those obtained from chamois- or pyrenean ibex-specific systems, serving as endogenous control for the total content of PCR-amplifiable DNA in the sample. Analysis of experimental raw and heat-treated binary mixtures of chamois and pyrenean ibex meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target DNAs in the range of 0.1-0.8%, depending on the species and treatment of the meat samples.

  19. Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

    Directory of Open Access Journals (Sweden)

    Waight Pauline A

    2006-04-01

    Full Text Available Abstract Background/Aim The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials. Method Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers. Results The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 × 103 to 1.5 × 108 copies/mL and correlated well with those from a commercial assay (r = 0.91, (p Conclusion This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

  20. Gene-targeted embryonic stem cells: real-time PCR assay for estimation of the number of neomycin selection cassettes

    Directory of Open Access Journals (Sweden)

    Mancini Cecilia

    2011-10-01

    Full Text Available Abstract In the preparation of transgenic murine ES cells it is important to verify the construct has a single insertion, because an ectopic neomycin phosphortransferase positive selection cassette (NEO may cause a position effect. During a recent work, where a knockin SCA28 mouse was prepared, we developed two assays based on Real-Time PCR using both SYBR Green and specific minor groove binder (MGB probes to evaluate the copies of NEO using the comparative delta-delta Ct method versus the Rpp30 reference gene. We compared the results from Southern blot, routinely used to quantify NEO copies, with the two Real-Time PCR assays. Twenty-two clones containing the single NEO copy showed values of 0.98 ± 0.24 (mean ± 2 S.D., and were clearly distinguishable from clones with two or more NEO copies. This method was found to be useful, easy, sensitive and fast and could substitute for the widely used, but laborious Southern blot method.

  1. Development of a novel quantitative real-time assay using duplex mutation primers for rapid detection of Candida species.

    Science.gov (United States)

    Xia, Qian-Feng; Liu, Jin-Bo; Liu, Ping; Qin, Xi; Qian, Shi-Yun; Tu, Zhi-Guang

    2012-01-01

    We developed a novel quantitative real-time PCR for quantitating Candida DNA based on the duplex mutation primer principle, in which a signal is generated by melting a duplex mutation primer during renaturation. The duplex mutation primers are much more specific than double-stranded DNA dyes such as SYBR-Green I and, unlike other probes, do not require the double-labeled synthesis of fluorophore and a quencher on the same molecule. A total of 176 clinical blood specimens were obtained from patients hospitalized in our hospital with clinically proven or suspected systemic Candida infection. The presence of DNA from pathogens in the Candida species was detected using real-time PCR targeting of an internal transcribed spacer region of a fungal gene. The assay exhibited a low limit of detection (10 CFU/ml of blood), an excellent reproducibility and specificity. Twenty-eight positive samples exhibited a wide range of Candida species loads, extending from 13 to 90,528 CFU/ml of blood. The sensitivity and specificity of the present assay were 100 and 97.4%, respectively, compared with the results of blood culture. Our data suggest that this assay may be appropriate for use in clinical laboratories as a simple, low-cost and rapid screening test for the most frequently encountered Candida species.

  2. Real-Time PCR Detection ofBurkholderia cepaciain Pharmaceutical Products Contaminated with Low Levels of Bacterial Contamination.

    Science.gov (United States)

    Jimenez, Luis; Jashari, Theranda; Vasquez, Jenifer; Zapata, Stephanie; Bochis, Joy; Kulko, Margarita; Ellman, Victoria; Gardner, Matthew; Choe, Tina

    2018-01-01

    A real-time polymerase chain reaction (RT-PCR) assay was developed to detect Burkholderia cepacia in pharmaceutical products contaminated with low levels of bacteria. Different pharmaceutical suspensions were artificially contaminated with B. cepacia , Escherichia coli , Staphylococcus aureus , and Bacillus megaterium After a 24 h incubation in trypticase soy broth with Tween 20, samples were streaked on mannitol salt, phenyl ethyl alcohol, eosin methylene blue, MacConkey, and pseudomonas isolation agar. Microbial DNA was extracted from each sample by using a Tris-EDTA, proteinase K, Tween 20 buffer. Regular PCR targeting the 1.5 kilobases 16S rRNA eubacterial gene and cloning showed the predominant DNA in the extracted mix belonged to E. coli Selective media isolation of bacterial contamination showed B. cepacia only detected on pseudomonas isolation while eosin methylene blue and MacConkey detected only E. coli RT-PCR using primers PSL1 and PSR1 amplified a 209 bp 16S rRNA fragment using a Roche LightCycler 96 ® system with SYBR green I, a common double-stranded binding dye. The cycle at which fluorescence from amplification exceeds the background fluorescence was referred to as quantification cycle. All samples were found to be positive by standard microbiological testing and RT-PCR. B. cepacia was detected within 30 h in all contaminated samples using RT-PCR. Based upon standard curve analysis of B. cepacia DNA, the minimum DNA concentration that could be detected was 10 fg/uL with a correlation value of 0.98. RT-PCR detection of B. cepacia allowed faster quality control analysis, corrective actions, and process optimization. LAY ABSTRACT: A real-time polymerase chain reaction (RT-PCR) assay was developed to detect Burkholderia cepacia in pharmaceutical products contaminated with low levels of bacteria. B. cepacia is the number one reason for microbial contamination recalls of non-sterile drug products in the USA. RT-PCR using primers PSL1 and PSR1 amplified a

  3. Real exchange rate misalignments

    OpenAIRE

    Terra, Maria Cristina T.; Valladares, Frederico Estrella Carneiro

    2003-01-01

    This paper characterizes episodes of real appreciations and depreciations for a sample of 85 countries, approximately from 1960 to 1998. First, the equilibrium real exchange rate series are constructed for each country using Goldfajn and Valdes (1999) methodology (cointegration with fundamentals). Then, departures from equilibrium real exchange rate (misalignments) are obtained, and a Markov Switching Model is used to characterize the misalignments series as stochastic autor...

  4. Elements of real analysis

    CERN Document Server

    Sprecher, David A

    2010-01-01

    This classic text in introductory analysis delineates and explores the intermediate steps between the basics of calculus and the ultimate stage of mathematics: abstraction and generalization.Since many abstractions and generalizations originate with the real line, the author has made it the unifying theme of the text, constructing the real number system from the point of view of a Cauchy sequence (a step which Dr. Sprecher feels is essential to learn what the real number system is).The material covered in Elements of Real Analysis should be accessible to those who have completed a course in

  5. Real-time LightCycler polymerase chain reaction and melting temperature analysis for identification of clinically important Candida spp.

    Science.gov (United States)

    Khan, Ziauddin; Mustafa, Abu Salim; Alam, Fasahat Fakhar

    2009-08-01

    Invasive candidiasis is a major fungal infection occurring in patients who have prolonged hospital admissions. The rapid detection and confirmation of Candida spp. in clinical specimens is essential for efficient management and improved prognosis of these patients. The purpose of this study was to develop a real-time LightCycler polymerase chain reaction (PCR) assay for the identification of Candida spp. commonly associated with invasive infections. Using the LightCycler PCR System, the targets of genomic DNA isolated from the reference strains of 6 Candida spp. were amplified using genus- and species-specific primers, and detected in real-time employing SYBR Green fluorescent dye. The identity of Candida spp. was established by melting curve analysis. A similar analysis was performed with clinical isolates (n = 72) previously identified by conventional methods. The melting curve analysis of amplified DNA from the reference strains could differentiate between Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata, Candida krusei, and Candida dubliniensis. The specificity of the real-time PCR assay was validated by testing 72 clinical isolates of Candida spp. with 100% concordance, as compared with conventional identification methods. The notable findings of the study were differentiation of C. krusei from all other Candida spp. tested and of C. dubliniensis from C. albicans by melting temperature analysis; the latter 2 species share common phenotypic characteristics of germ-tube formation and chlamydospore production, so are often misidentified. Real-time PCR using LightCycler and melting curve analysis are reliable methods for rapid identification of 6 Candida spp. frequently associated with candidemia and invasive candidiasis.

  6. Analytic real algebras.

    Science.gov (United States)

    Seo, Young Joo; Kim, Young Hee

    2016-01-01

    In this paper we construct some real algebras by using elementary functions, and discuss some relations between several axioms and its related conditions for such functions. We obtain some conditions for real-valued functions to be a (edge) d -algebra.

  7. Real Time Systems

    DEFF Research Database (Denmark)

    Christensen, Knud Smed

    2000-01-01

    Describes fundamentals of parallel programming and a kernel for that. Describes methods for modelling and checking parallel problems. Real time problems.......Describes fundamentals of parallel programming and a kernel for that. Describes methods for modelling and checking parallel problems. Real time problems....

  8. Real Topological Cyclic Homology

    DEFF Research Database (Denmark)

    Høgenhaven, Amalie

    The main topics of this thesis are real topological Hochschild homology and real topological cyclic homology. If a ring or a ring spectrum is equipped with an anti-involution, then it induces additional structure on the topological Hochschild homology spectrum. The group O(2) acts on the spectrum......, where O(2) is the semi-direct product of T, the multiplicative group of complex number of modulus 1, by the group G=Gal(C/R). We refer to this O(2)-spectrum as the real topological Hochschild homology. This generalization leads to a G-equivariant version of topological cyclic homology, which we call...... real topological cyclic homology. The first part of the thesis computes the G-equivariant homotopy type of the real topological cyclic homology of spherical group rings at a prime p with anti-involution induced by taking inverses in the group. The second part of the thesis investigates the derived G...

  9. Basic real analysis

    CERN Document Server

    Sohrab, Houshang H

    2014-01-01

    This expanded second edition presents the fundamentals and touchstone results of real analysis in full rigor, but in a style that requires little prior familiarity with proofs or mathematical language. The text is a comprehensive and largely self-contained introduction to the theory of real-valued functions of a real variable. The chapters on Lebesgue measure and integral have been rewritten entirely and greatly improved. They now contain Lebesgue’s differentiation theorem as well as his versions of the Fundamental Theorem(s) of Calculus. With expanded chapters, additional problems, and an expansive solutions manual, Basic Real Analysis, Second Edition, is ideal for senior undergraduates and first-year graduate students, both as a classroom text and a self-study guide. Reviews of first edition: The book is a clear and well-structured introduction to real analysis aimed at senior undergraduate and beginning graduate students. The prerequisites are few, but a certain mathematical sophistication is required. ....

  10. From real life to real life

    DEFF Research Database (Denmark)

    Svalgaard, Lotte

    2017-01-01

    In Action Learning programmes, it is held central to work on real business challenges (task) while learning about team and self (process); staying mindful aware of the process is referred to in this paper as ‘double awareness’, and emphasises noticing and acting on process cues while working...... on the task. As business challenges within Action Learning programmes are real, pertinent, and worked with in the context of the organisation, implementation of potential solutions to the challenges is proved to be efficient. However, less is known of how individual participants manage to stay with double...... awareness upon re-entry into the organisational routines left behind. The aim of this paper is to explore when, why, and how participants manage to maintain double awareness and act on what they notice when back in organisational reality – and what characterise the moments where they struggle to do so...

  11. Targeting a polyketide synthase gene for Aspergillus carbonarius quantification and ochratoxin A assessment in grapes using real-time PCR

    International Nuclear Information System (INIS)

    Atoui, A.; Mathieu, F.; Lebrihi, A.

    2007-01-01

    Aspergillus carbonarius is an ochratoxin producing fungus that has been considered to be responsible of the ochratoxin A (OTA) contamination in grapes and wine. In order to monitor and quantify A. carbonarius, a specific primer pair Ac12RL O TAF/Ac12RL O TAR has been designed from the acyltransferase (AT) domain of the polyketide synthase sequence Ac12RL3 to amplify 141 bp PCR product. Among the mycotoxigenic fungi tested, only A. carbonarius gave a positive result. This specific primer pair was also successfully employed in real-time PCR conjugated with SYBR Green I dye for the direct quantification of this fungus in grape samples. A positive correlation (R2 = 0.81) was found between A. carbonarius DNA content and OTA concentration in 72 grape samples, allowing for the estimation of the potential risk from OTA contamination. Consequently, this work offers a quick alternative to conventional methods of OTA quantification and mycological detection and quantification of A. carbonarius in grapes. (author)

  12. Evaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes

    Directory of Open Access Journals (Sweden)

    Lilian Giotto Zaros

    2010-01-01

    Full Text Available Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO, glyceraldehyde 3-phosphate dehydrogenase (GAPDH and succinate dehydrogenase complex subunit A (SDHA, were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and was ranked as the most stable in abomasum and lymph node tissues. On the other hand, SDHA was the most stable in the small intestine followed by RPLO and GAPDH. These findings demonstrate the importance of correctly choosing reference genes prior to relative quantification. In addition, we determined that reference genes used in sheep from temperate regions, when properly tested, can be applied in animals from tropical regions such as the Brazilian Somalis sheep.

  13. Introduction to real analysis

    CERN Document Server

    Schramm, Michael J

    2008-01-01

    This text forms a bridge between courses in calculus and real analysis. It focuses on the construction of mathematical proofs as well as their final content. Suitable for upper-level undergraduates and graduate students of real analysis, it also provides a vital reference book for advanced courses in mathematics.The four-part treatment begins with an introduction to basic logical structures and techniques of proof, including discussions of the cardinality concept and the algebraic and order structures of the real and rational number systems. Part Two presents in-depth examinations of the compl

  14. Real Option Analysis (ROA

    Directory of Open Access Journals (Sweden)

    Armanto Witjaksono

    2003-03-01

    Full Text Available Net Present Value (NPV method have populer since middle 70’s and now most of expert felt that method has several limitation, especially if used to analyse big scale investment alocation capital. Another method that begin to popular is Real Option Analysis (ROA that use to replace Net Present Value (NPV method. The strenght of Real Option Analysis (ROA method is the flexiblelity in giving information for the decision maker. The weakness of Real Option Analysis (ROA method is the simple mathematic formula, as the formula in NPV method, is not found yet. 

  15. Real-time shadows

    CERN Document Server

    Eisemann, Elmar; Assarsson, Ulf; Wimmer, Michael

    2011-01-01

    Important elements of games, movies, and other computer-generated content, shadows are crucial for enhancing realism and providing important visual cues. In recent years, there have been notable improvements in visual quality and speed, making high-quality realistic real-time shadows a reachable goal. Real-Time Shadows is a comprehensive guide to the theory and practice of real-time shadow techniques. It covers a large variety of different effects, including hard, soft, volumetric, and semi-transparent shadows.The book explains the basics as well as many advanced aspects related to the domain

  16. Making It Real

    Science.gov (United States)

    McCartney, Robert; Tenenberg, Josh

    2008-01-01

    Some have proposed that realistic problem situations are better for learning. This issue contains two articles that examine the effects of "making it real" in computer architecture and human-computer interaction.

  17. Real Property Tax - 2016

    Data.gov (United States)

    Montgomery County of Maryland — This data represents all of the County’s residential real estate properties and all of the associated tax charges and credits with that property processed at the...

  18. Real Property Tax - 2017

    Data.gov (United States)

    Montgomery County of Maryland — This data represents all of the County’s residential real estate properties and all of the associated tax charges and credits with that property processed at the...

  19. Real Property Tax Rates

    Data.gov (United States)

    Montgomery County of Maryland — The Levy Year 2012 real property tax rate dataset reflects all the rates per $100 set each year by the County Council. These rates are applied to the assessed value...

  20. [Establish and optimization of real-time fluorescent reverse transcription loop-mediated isothermal amplification for detection of avian influenza H5 hemagglutinin gene].

    Science.gov (United States)

    Liu, Yun; Tang, Jin-ming; Tao, Hong; Sun, Jie; Lu, Ti-kang; Liao, Li-shan; Liu, Jian-li; Zeng, Shao-ling; Cao, Chen-fu; Zhang, Cai-hong; Ruan, Zhou-xi; Lv, Jian-qiang; Yang, Jun-xing; Hua, Qun-yi; Chen, Zheng-li; Qin, Zhi-feng

    2013-09-01

    H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye. The results showed the sensitivity of this method was 100 times higher than that of standard real-time fluorescent RT-PCR, and the detection limit was one copy of the gene per reaction. This method had no cross-reactivity with other common avian respiratory tract infectious disease-related pathogens such as IBV and NDV. The present study suggested Calcein was the optimum dye. Small-scale tests suggested this method was reliable for survey monitoring of AIV-H5 on the spot, indicating its potential applications in field investigation.

  1. Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola.

    Science.gov (United States)

    Taverniers, Isabel; Windels, Pieter; Vaïtilingom, Marc; Milcamps, Anne; Van Bockstaele, Erik; Van den Eede, Guy; De Loose, Marc

    2005-04-20

    Since the 18th of April 2004, two new regulations, EC/1829/2003 on genetically modified food and feed products and EC/1830/2003 on traceability and labeling of GMOs, are in force in the EU. This new, comprehensive regulatory framework emphasizes the need of an adequate tracing system. Unique identifiers, such as the transgene genome junction region or a specific rearrangement within the transgene DNA, should form the basis of such a tracing system. In this study, we describe the development of event-specific tracing systems for transgenic maize lines Bt11, Bt176, and GA21 and for canola event GT73. Molecular characterization of the transgene loci enabled us to clone an event-specific sequence into a plasmid vector, to be used as a marker, and to develop line-specific primers. Primer specificity was tested through qualitative PCRs and dissociation curve analysis in SYBR Green I real-time PCRs. The primers were then combined with event-specific TaqMan probes in quantitative real-time PCRs. Calibration curves were set up both with genomic DNA samples and the newly synthesized plasmid DNA markers. It is shown that cloned plasmid GMO target sequences are perfectly suitable as unique identifiers and quantitative calibrators. Together with an event-specific primer pair and a highly specific TaqMan probe, the plasmid markers form crucial components of a unique and straighforward tracing system for Bt11, Bt176, and GA21 maize and GT73 canola events.

  2. Synthesis of O-serogroup specific positive controls and real-time PCR standards for nine clinically relevant non-O157 STECs.

    Science.gov (United States)

    Conrad, Cheyenne C; Gilroyed, Brandon H; McAllister, Tim A; Reuter, Tim

    2012-10-01

    Non-O157 Shiga toxin producing Escherichia coli (STEC) are gaining recognition as human pathogens, but no standardized method exists to identify them. Sequence analysis revealed that STEC can be classified on the base of variable O antigen regions into different O serotypes. Polymerase chain reaction is a powerful technique for thorough screening and complex diagnosis for these pathogens, but requires a positive control to verify qualitative and/or quantitative DNA-fragment amplification. Due to the pathogenic nature of STEC, controls are not readily available and cell culturing of STEC reference strains requires biosafety conditions of level 2 or higher. In order to bypass this limitation, controls of stacked O-type specific DNA-fragments coding for primer recognition sites were designed to screen for nine STEC serotypes frequently associated with human infection. The synthetic controls were amplified by PCR, cloned into a plasmid vector and transferred into bacteria host cells. Plasmids amplified by bacterial expression were purified, serially diluted and tested as standards for real-time PCR using SYBR Green and TaqMan assays. Utility of synthetic DNA controls was demonstrated in conventional and real-time PCR assays and validated with DNA from natural STEC strains. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Real-time polymerase chain reaction optimised for hepatitis C virus detection in dried blood spots from HIV-exposed infants, KwaZulu-Natal, South Africa

    Directory of Open Access Journals (Sweden)

    Anneta Naidoo

    2016-03-01

    Objectives: The aim of this study was to optimise a real-time HCV PCR method to detect HCV RNA from infant DBS specimens for use as a tool for HCV surveillance in KwaZulu-Natal, South Africa. Method: The LightCycler® 2.0 instrument was used for the HCV PCR using the LightCycler® RNA Master SYBR Green I kit. Template volume, primer concentration and primer annealing temperatures were optimised and the method was used on 179 DBS specimens from HIV-exposed infants in KwaZulu-Natal. Results: Primer concentrations adjusted to 0.25 µM and a template volume of 10 µL improved the PCR amplification. Primer annealing temperatures lowered from 65 °C to 58 °C resulted in higher quantities of amplified PCR product. The limit of detection of the optimised HCV PCR assay was between 1200 IU/mL and 3580 IU/mL of HCV RNA. HCV was not detected in any of the 179 DBS specimens. Conclusion: The optimised real-time HCV PCR on infant DBS specimens performed well, but HCV was not found in this surveillance study. HIV infection may have little impact on the vertical transmission of HCV in this region.

  4. A One-Step Real-Time Multiplex PCR for Screening Y-Chromosomal Microdeletions without Downstream Amplicon Size Analysis

    Science.gov (United States)

    Kozina, Viviana; Cappallo-Obermann, Heike; Gromoll, Jörg; Spiess, Andrej-Nikolai

    2011-01-01

    Backgound Y-chromosomal microdeletions (YCMD) are one of the major genetic causes for non-obstructive azoospermia. Genetic testing for YCMD by multiplex polymerase chain reaction (PCR) is an established method for quick and robust screening of deletions in the AZF regions of the Y-chromosome. Multiplex PCRs have the advantage of including a control gene in every reaction and significantly reducing the number of reactions needed to screen the relevant genomic markers. Principal Findings The widely established “EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions (2004)” were used as a basis for designing a real-time multiplex PCR system, in which the YCMD can simply be identified by their melting points. For this reason, some AZF primers were substituted by primers for regions in their genomic proximity, and the ZFX/ZFY control primer was exchanged by the AMELX/AMELY control primer. Furthermore, we substituted the classical SybrGreen I dye by the novel and high-performing DNA-binding dye EvaGreen™ and put substantial effort in titrating the primer combinations in respect to optimal melting peak separation and peak size. Significance With these changes, we were able to develop a platform-independent and robust real-time based multiplex PCR, which makes the need for amplicon identification by electrophoretic sizing expendable. By using an open-source system for real-time PCR analysis, we further demonstrate the applicability of automated melting point and YCMD detection. PMID:21887237

  5. Acetobacter malorum and Acetobacter cerevisiae identification and quantification by Real-Time PCR with TaqMan-MGB probes.

    Science.gov (United States)

    Valera, Maria José; Torija, Maria Jesús; Mas, Albert; Mateo, Estibaliz

    2013-10-01

    The identification and quantification of Acetobacter malorum and Acetobacter cerevisiae in wine and vinegar were performed using the Real-Time PCR (RT-PCR) with two TaqMan-MGB probes designed to amplify the internal transcribed spacer (ITS) region between the 16S-23S rRNA genes. The primers and probes were highly specific, with a detection limit of 10² cells/ml for both species, and the efficiency of the technique was >80%. The RT-PCR technique with these two new TaqMan-MGB probes, together with the five (Acetobacter aceti, Acetobacter pasteurianus, Gluconobacter oxydans, Gluconacetobacter hansenii and Gluconacetobacter europaeus) that are already available (Torija et al., 2010), were validated on known concentrations of Acetic Acid Bacteria (AAB) grown in glucose medium (GY) and in inoculated matrices of wine and vinegar. Furthermore, this technique was applied to evaluate the AAB population in real wine samples collected in the Canary Islands. PCR enrichment performed prior to RT-PCR increased the accuracy of quantification and produced results similar to those detected with SYBR-Green. In real wine samples, the total AAB enumeration ranged from 9 × 10² to 10⁶ cells/ml, and the seven AAB species tested were detected in more than one sample. However, AAB recovery on plates was poor; the isolates obtained on plates were A. malorum, G. oxydans, A. cerevisiae and A. pasteurianus species. RT-PCR with TaqMan-MGB probes is an accurate, specific and fast method for the identification and quantification of AAB species commonly found in wine and vinegar. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Real time expert systems

    International Nuclear Information System (INIS)

    Asami, Tohru; Hashimoto, Kazuo; Yamamoto, Seiichi

    1992-01-01

    Recently, aiming at the application to the plant control for nuclear reactors and traffic and communication control, the research and the practical use of the expert system suitable to real time processing have become conspicuous. In this report, the condition for the required function to control the object that dynamically changes within a limited time is presented, and the technical difference between the real time expert system developed so as to satisfy it and the expert system of conventional type is explained with the actual examples and from theoretical aspect. The expert system of conventional type has the technical base in the problem-solving equipment originating in STRIPS. The real time expert system is applied to the fields accompanied by surveillance and control, to which conventional expert system is hard to be applied. The requirement for the real time expert system, the example of the real time expert system, and as the techniques of realizing real time processing, the realization of interruption processing, dispersion processing, and the mechanism of maintaining the consistency of knowledge are explained. (K.I.)

  7. Real parabolic vector bundles over a real curve

    Indian Academy of Sciences (India)

    by Seshadri [4] and their moduli studied in [2]. Here we consider real vector bundles over a real curve and define parabolic structures on real vector bundles. By a real curve, we mean a pair (X,σX ), where X is a compact Riemann surface and. σX is an anti-holomorphic involution on X. A real vector bundle over a real curve ...

  8. 1,2-propanediol-trehalose mixture as a potent quantitative real-time PCR enhancer

    Directory of Open Access Journals (Sweden)

    Dráberová Lubica

    2011-04-01

    Full Text Available Abstract Background Quantitative real-time PCR (qPCR is becoming increasingly important for DNA genotyping and gene expression analysis. For continuous monitoring of the production of PCR amplicons DNA-intercalating dyes are widely used. Recently, we have introduced a new qPCR mix which showed improved amplification of medium-size genomic DNA fragments in the presence of DNA dye SYBR green I (SGI. In this study we tested whether the new PCR mix is also suitable for other DNA dyes used for qPCR and whether it can be applied for amplification of DNA fragments which are difficult to amplify. Results We found that several DNA dyes (SGI, SYTO-9, SYTO-13, SYTO-82, EvaGreen, LCGreen or ResoLight exhibited optimum qPCR performance in buffers of different salt composition. Fidelity assays demonstrated that the observed differences were not caused by changes in Taq DNA polymerase induced mutation frequencies in PCR mixes of different salt composition or containing different DNA dyes. In search for a PCR mix compatible with all the DNA dyes, and suitable for efficient amplification of difficult-to-amplify DNA templates, such as those in whole blood, of medium size and/or GC-rich, we found excellent performance of a PCR mix supplemented with 1 M 1,2-propanediol and 0.2 M trehalose (PT enhancer. These two additives together decreased DNA melting temperature and efficiently neutralized PCR inhibitors present in blood samples. They also made possible more efficient amplification of GC-rich templates than betaine and other previously described additives. Furthermore, amplification in the presence of PT enhancer increased the robustness and performance of routinely used qPCRs with short amplicons. Conclusions The combined data indicate that PCR mixes supplemented with PT enhancer are suitable for DNA amplification in the presence of various DNA dyes and for a variety of templates which otherwise can be amplified with difficulty.

  9. Making fictions sound real

    DEFF Research Database (Denmark)

    Langkjær, Birger

    2010-01-01

    of sounds. Apart from an account of the kinds of sounds that typically appear in a specific genre, a genre analysis of sound may also benefit from a functionalist approach that focuses on how sounds can make both realist and non-realist aspects of genres sound real to audiences.......This article examines the role that sound plays in making fictions perceptually real to film audiences, whether these fictions are realist or non-realist in content and narrative form. I will argue that some aspects of film sound practices and the kind of experiences they trigger are related...... to basic rules of human perception, whereas others are more properly explained in relation to how aesthetic devices, including sound, are used to characterise the fiction and thereby make it perceptually real to its audience. Finally, I will argue that not all genres can be defined by a simple taxonomy...

  10. Essential real analysis

    CERN Document Server

    Field, Michael

    2017-01-01

    This book provides a rigorous introduction to the techniques and results of real analysis, metric spaces and multivariate differentiation, suitable for undergraduate courses. Starting from the very foundations of analysis, it offers a complete first course in real analysis, including topics rarely found in such detail in an undergraduate textbook such as the construction of non-analytic smooth functions, applications of the Euler-Maclaurin formula to estimates, and fractal geometry.  Drawing on the author’s extensive teaching and research experience, the exposition is guided by carefully chosen examples and counter-examples, with the emphasis placed on the key ideas underlying the theory. Much of the content is informed by its applicability: Fourier analysis is developed to the point where it can be rigorously applied to partial differential equations or computation, and the theory of metric spaces includes applications to ordinary differential equations and fractals. Essential Real Analysis will appeal t...

  11. Real mathematical analysis

    CERN Document Server

    Pugh, Charles C

    2015-01-01

    Based on an honors course taught by the author at UC Berkeley, this introduction to undergraduate real analysis gives a different emphasis by stressing the importance of pictures and hard problems. Topics include: a natural construction of the real numbers, four-dimensional visualization, basic point-set topology, function spaces, multivariable calculus via differential forms (leading to a simple proof of the Brouwer Fixed Point Theorem), and a pictorial treatment of Lebesgue theory. Over 150 detailed illustrations elucidate abstract concepts and salient points in proofs. The exposition is informal and relaxed, with many helpful asides, examples, some jokes, and occasional comments from mathematicians, such as Littlewood, Dieudonné, and Osserman. This book thus succeeds in being more comprehensive, more comprehensible, and more enjoyable, than standard introductions to analysis. New to the second edition of Real Mathematical Analysis is a presentation of Lebesgue integration done almost entirely using the un...

  12. Making fictions sound real

    DEFF Research Database (Denmark)

    Langkjær, Birger

    2010-01-01

    This article examines the role that sound plays in making fictions perceptually real to film audiences, whether these fictions are realist or non-realist in content and narrative form. I will argue that some aspects of film sound practices and the kind of experiences they trigger are related...... to basic rules of human perception, whereas others are more properly explained in relation to how aesthetic devices, including sound, are used to characterise the fiction and thereby make it perceptually real to its audience. Finally, I will argue that not all genres can be defined by a simple taxonomy...... of sounds. Apart from an account of the kinds of sounds that typically appear in a specific genre, a genre analysis of sound may also benefit from a functionalist approach that focuses on how sounds can make both realist and non-realist aspects of genres sound real to audiences....

  13. Real-time radiography

    International Nuclear Information System (INIS)

    Bossi, R.H.; Oien, C.T.

    1981-01-01

    Real-time radiography is used for imaging both dynamic events and static objects. Fluorescent screens play an important role in converting radiation to light, which is then observed directly or intensified and detected. The radiographic parameters for real-time radiography are similar to conventional film radiography with special emphasis on statistics and magnification. Direct-viewing fluoroscopy uses the human eye as a detector of fluorescent screen light or the light from an intensifier. Remote-viewing systems replace the human observer with a television camera. The remote-viewing systems have many advantages over the direct-viewing conditions such as safety, image enhancement, and the capability to produce permanent records. This report reviews real-time imaging system parameters and components

  14. Modern real analysis

    CERN Document Server

    Ziemer, William P

    2017-01-01

    This first year graduate text is a comprehensive resource in real analysis based on a modern treatment of measure and integration. Presented in a definitive and self-contained manner, it features a natural progression of concepts from simple to difficult. Several innovative topics are featured, including differentiation of measures, elements of Functional Analysis, the Riesz Representation Theorem, Schwartz distributions, the area formula, Sobolev functions and applications to harmonic functions. Together, the selection of topics forms a sound foundation in real analysis that is particularly suited to students going on to further study in partial differential equations. This second edition of Modern Real Analysis contains many substantial improvements, including the addition of problems for practicing techniques, and an entirely new section devoted to the relationship between Lebesgue and improper integrals. Aimed at graduate students with an understanding of advanced calculus, the text will also appeal to mo...

  15. Real analysis and probability

    CERN Document Server

    Ash, Robert B; Lukacs, E

    1972-01-01

    Real Analysis and Probability provides the background in real analysis needed for the study of probability. Topics covered range from measure and integration theory to functional analysis and basic concepts of probability. The interplay between measure theory and topology is also discussed, along with conditional probability and expectation, the central limit theorem, and strong laws of large numbers with respect to martingale theory.Comprised of eight chapters, this volume begins with an overview of the basic concepts of the theory of measure and integration, followed by a presentation of var

  16. Real parabolic vector bundles over a real curve

    Indian Academy of Sciences (India)

    Abstract. We define real parabolic structures on real vector bundles over a real curve. Let (X,σX ) be a real curve, and let S ⊂ X be a non-empty finite subset of X such that σX (S) = S. Let N ≥ 2 be an integer. We construct an N-fold cyclic cover p : Y → X in the category of real curves, ramified precisely over each point of S, ...

  17. Real Compact Surfaces

    Indian Academy of Sciences (India)

    The classification of real compact surfaces is a main result which is at the same time easy to understand and non- trivial, simple in formulation and rich in consequences. The aim of this article is to explain the theorem by means of many drawings. It is an invitation to a visual approach of mathematics. First Definitions and ...

  18. Modelling Real Property Transactions

    DEFF Research Database (Denmark)

    Stubkjær, Erik

    2003-01-01

    Der er store forskelle i financieringsomkostningerne ved hussalg i forskellige lande. Er disse forskelle nødvendige? og kan udgifterne reduceres? Sådanne spørgsmål søger forskningsprojektet ?Modelling Real Property Transactions?, officielt sat i værk indenfor rammerne af COST (European Co...

  19. Real Estate in Denmark

    DEFF Research Database (Denmark)

    Rimmer, Nina Røhr

    2016-01-01

    A general introduction to buying, owning and selling Property i Denmark. The cost of buying and selling property and the tax implications as well as the general concept of real estate is introduced. The paper includes also a brief introduction to some of the aspects which you should beware of when...

  20. Un-real Estate

    DEFF Research Database (Denmark)

    Ulfstjerne, Michael Alexander

    Based on ethnographic fieldwork in Ordos China the dissertation ‘Un-real Estate’ traces a local economic boom into its subsequent bust. It gives an account of how state as well as non-state actors appropriate state development schemes and the unintended effects resulting from the colluding forces...

  1. Is Optimism Real?

    Science.gov (United States)

    Simmons, Joseph P.; Massey, Cade

    2012-01-01

    Is optimism real, or are optimistic forecasts just cheap talk? To help answer this question, we investigated whether optimistic predictions persist in the face of large incentives to be accurate. We asked National Football League football fans to predict the winner of a single game. Roughly half (the partisans) predicted a game involving their…

  2. Being Real for Children.

    Science.gov (United States)

    Carter, Margie

    1995-01-01

    Because ready role models for today's children are media-created superheroes and celebrities of television and film, children need real-life role models who guide them into realistic personal and social pathways. As principal adult contacts, teachers can be such role models. Specific strategies for encouraging teachers in this role are presented.…

  3. Introductory real analysis

    CERN Document Server

    Kolmogorov, A N; Silverman, Richard A

    1975-01-01

    Self-contained and comprehensive, this elementary introduction to real and functional analysis is readily accessible to those with background in advanced calculus. It covers basic concepts and introductory principles in set theory, metric spaces, topological and linear spaces, linear functionals and linear operators, and much more. 350 problems. 1970 edition.

  4. Real World Europeanisation

    DEFF Research Database (Denmark)

    Pedersen, Morten Jarlbæk

    2017-01-01

    Several studies of the impacts of the EU on modern policies and polities evade investigation of the most detailed level of Europeanisation, i.e. gradual legal changes. This seems odd as it is exactly these small gears turning that constitute perhaps the most real and concrete of all types of Euro...

  5. A Java program for LRE-based real-time qPCR that enables large-scale absolute quantification.

    Science.gov (United States)

    Rutledge, Robert G

    2011-03-02

    Linear regression of efficiency (LRE) introduced a new paradigm for real-time qPCR that enables large-scale absolute quantification by eliminating the need for standard curves. Developed through the application of sigmoidal mathematics to SYBR Green I-based assays, target quantity is derived directly from fluorescence readings within the central region of an amplification profile. However, a major challenge of implementing LRE quantification is the labor intensive nature of the analysis. Utilizing the extensive resources that are available for developing Java-based software, the LRE Analyzer was written using the NetBeans IDE, and is built on top of the modular architecture and windowing system provided by the NetBeans Platform. This fully featured desktop application determines the number of target molecules within a sample with little or no intervention by the user, in addition to providing extensive database capabilities. MS Excel is used to import data, allowing LRE quantification to be conducted with any real-time PCR instrument that provides access to the raw fluorescence readings. An extensive help set also provides an in-depth introduction to LRE, in addition to guidelines on how to implement LRE quantification. The LRE Analyzer provides the automated analysis and data storage capabilities required by large-scale qPCR projects wanting to exploit the many advantages of absolute quantification. Foremost is the universal perspective afforded by absolute quantification, which among other attributes, provides the ability to directly compare quantitative data produced by different assays and/or instruments. Furthermore, absolute quantification has important implications for gene expression profiling in that it provides the foundation for comparing transcript quantities produced by any gene with any other gene, within and between samples.

  6. A Java program for LRE-based real-time qPCR that enables large-scale absolute quantification.

    Directory of Open Access Journals (Sweden)

    Robert G Rutledge

    Full Text Available BACKGROUND: Linear regression of efficiency (LRE introduced a new paradigm for real-time qPCR that enables large-scale absolute quantification by eliminating the need for standard curves. Developed through the application of sigmoidal mathematics to SYBR Green I-based assays, target quantity is derived directly from fluorescence readings within the central region of an amplification profile. However, a major challenge of implementing LRE quantification is the labor intensive nature of the analysis. FINDINGS: Utilizing the extensive resources that are available for developing Java-based software, the LRE Analyzer was written using the NetBeans IDE, and is built on top of the modular architecture and windowing system provided by the NetBeans Platform. This fully featured desktop application determines the number of target molecules within a sample with little or no intervention by the user, in addition to providing extensive database capabilities. MS Excel is used to import data, allowing LRE quantification to be conducted with any real-time PCR instrument that provides access to the raw fluorescence readings. An extensive help set also provides an in-depth introduction to LRE, in addition to guidelines on how to implement LRE quantification. CONCLUSIONS: The LRE Analyzer provides the automated analysis and data storage capabilities required by large-scale qPCR projects wanting to exploit the many advantages of absolute quantification. Foremost is the universal perspective afforded by absolute quantification, which among other attributes, provides the ability to directly compare quantitative data produced by different assays and/or instruments. Furthermore, absolute quantification has important implications for gene expression profiling in that it provides the foundation for comparing transcript quantities produced by any gene with any other gene, within and between samples.

  7. Development of real-time PCR array for simultaneous detection of eight human blood-borne viral pathogens.

    Directory of Open Access Journals (Sweden)

    Natalia Pripuzova

    Full Text Available BACKGROUND: Real-time PCR array for rapid detection of multiple viral pathogens should be highly useful in cases where the sample volume and the time of testing are limited, i.e. in the eligibility testing of tissue and organ donors. FINDINGS: We developed a real-time PCR array capable of simultaneously detecting eight human viral pathogens: human immunodeficiency virus types 1 and 2 (HIV-1 and -2, hepatitis B virus (HBV, hepatitis C virus (HCV, human T-cell leukemia virus-1 and -2 (HTLV-1 and -2, vaccinia virus (VACV and West Nile virus (WNV. One hundred twenty (120 primers were designed using a combination of bioinformatics approaches, and, after experimental testing, 24 primer sets targeting eight viral pathogens were selected to set up the array with SYBR Green chemistry. The specificity and sensitivity of the virus-specific primer sets selected for the array were evaluated using analytical panels with known amounts of viruses spiked into human plasma. The array detected: 10 genome equivalents (geq/ml of HIV-2 and HCV, 50 geq of HIV-1 (subtype B, HBV (genotype A and WNV. It detected 100-1,000 geq/ml of plasma of HIV-1 subtypes (A - G, group N and CRF (AE and AG isolates. Further evaluation with a panel consisting of 28 HIV-1 and HIV-2 clinical isolates revealed no cross-reactivity of HIV-1 or HIV-2 specific primers with another type of HIV. All 28 viral isolates were identified with specific primer sets targeting the most conserved genome areas. The PCR array correctly identified viral infections in a panel of 17 previously quantified clinical plasma samples positive for HIV-1, HCV or HBV at as low as several geq per PCR reaction. CONCLUSIONS: The viral array described here demonstrated adequate performance in the testing of donors' clinical samples. Further improvement in its sensitivity for the broad spectrum of HIV-1 subtypes is under development.

  8. Detection, quantification and genotyping of Herpes Simplex Virus in cervicovaginal secretions by real-time PCR: a cross sectional survey

    Directory of Open Access Journals (Sweden)

    Natividad-Sancho Angels

    2005-08-01

    Full Text Available Abstract Background Herpes Simplex Virus (HSV Genital Ulcer Disease (GUD is an important public health problem, whose interaction with HIV results in mutually enhancing epidemics. Conventional methods for detecting HSV tend to be slow and insensitive. We designed a rapid PCR-based assay to quantify and type HSV in cervicovaginal lavage (CVL fluid of subjects attending a Genito-Urinary Medicine (GUM clinic. Vaginal swabs, CVL fluid and venous blood were collected. Quantitative detection of HSV was conducted using real time PCR with HSV specific primers and SYBR Green I. Fluorogenic TaqMan Minor Groove Binder (MGB probes designed around a single base mismatch in the HSV DNA polymerase I gene were used to type HSV in a separate reaction. The Kalon test was used to detect anti-HSV-2 IgG antibodies in serum. Testing for HIV, other Sexually Transmitted Infections (STI and related infections was based on standard clinical and laboratory methods. Results Seventy consecutive GUM clinic attendees were studied. Twenty-seven subjects (39% had detectable HSV DNA in CVL fluid; HSV-2 alone was detected in 19 (70% subjects, HSV-1 alone was detected in 4 (15% subjects and both HSV types were detected in 4 (15% subjects. Eleven out of 27 subjects (41% with anti-HSV-2 IgG had detectable HSV-2 DNA in CVL fluid. Seven subjects (10% were HIV-positive. Three of seven (43% HIV-infected subjects and two of five subjects with GUD (40% were secreting HSV-2. None of the subjects in whom HSV-1 was detected had GUD. Conclusion Quantitative real-time PCR and Taqman MGB probes specific for HSV-1 or -2 were used to develop an assay for quantification and typing of HSV. The majority of subjects in which HSV was detected had low levels of CVL fluid HSV, with no detectable HSV-2 antibodies and were asymptomatic.

  9. On real quadric line complexes

    International Nuclear Information System (INIS)

    Krasnov, Vyacheslav A

    2010-01-01

    We describe the topological types of the real parts of the Kummer surfaces associated with real three-dimensional quadric line complexes. The topological type of the real part of such a surface is shown to depend on the number of real singular points: it is determined by the number of such points if any exist, and otherwise the real part of the Kummer surface is either empty or consists of one or two tori.

  10. Elements of real analysis

    CERN Document Server

    Al-Gwaiz, MA

    2006-01-01

    PREFACEPRELIMINARIES Sets FunctionsREAL NUMBERSField AxiomsOrder Axioms Natural Numbers, Integers, Rational NumbersCompleteness Axiom Decimal Representation of Real Numbers Countable SetsSEQUENCESSequences and ConvergenceProperties of Convergent Sequences Monotonic SequencesThe Cauchy Criterion Subsequences Upper and Lower Limits Open and Closed Sets INFINITE SERIESBasic Properties Convergence TestsLIMIT OF A FUNCTIONLimit of a Function Basic Theorems Some Extensions of the LimitMonotonic Functions CONTINUITY Continuous Functions Combinations of Continuous Functions Continuity on an IntervalUniformContinuityCompact Sets and ContinuityDIFFERENTIATION The DerivativeTheMean Value TheoremL'Hôpital's RuleTaylor's TheoremTHE RIEMANN INTEGRALRiemann Integrability Darboux's Theorem and Riemann SumsProperties of the Integral The Fundamental Theorem of Calculus Improper IntegralsSEQUENCES AND SERIES OF FUNCTIONSSequences of FunctionsProperties of Uniform ConvergenceSeries of FunctionsPower Series LEBESGUE MEASURE Clas...

  11. Route around real time

    International Nuclear Information System (INIS)

    Terrier, Francois

    1996-01-01

    The greater and greater autonomy and complexity asked to the control and command systems lead to work on introducing techniques such as Artificial Intelligence or concurrent object programming in industrial applications. However, while the critical feature of these systems impose to control the dynamics of the proposed solutions, their complexity often imposes a high adaptability to a partially modelled environment. The studies presented start from low level control and command systems to more complex applications at higher levels, such as 'supervision systems'. Techniques such as temporal reasoning and uncertainty management are proposed for the first studies, while the second are tackled with programming techniques based on the real time object paradigm. The outcomes of this itinerary crystallize on the ACCORD project which targets to manage - on the whole life cycle of a real time application - these two problematics, sometimes antagonistic: control of the dynamics and adaptivity. (author) [fr

  12. Real analysis on intervals

    CERN Document Server

    Choudary, A D R

    2014-01-01

    The book targets undergraduate and postgraduate mathematics students and helps them develop a deep understanding of mathematical analysis. Designed as a first course in real analysis, it helps students learn how abstract mathematical analysis solves mathematical problems that relate to the real world. As well as providing a valuable source of inspiration for contemporary research in mathematics, the book helps students read, understand and construct mathematical proofs, develop their problem-solving abilities and comprehend the importance and frontiers of computer facilities and much more. It offers comprehensive material for both seminars and independent study for readers with a basic knowledge of calculus and linear algebra. The first nine chapters followed by the appendix on the Stieltjes integral are recommended for graduate students studying probability and statistics, while the first eight chapters followed by the appendix on dynamical systems will be of use to students of biology and environmental scie...

  13. Real Snail Mail

    OpenAIRE

    Isley, Vicky; Smith, Paul; boredomresearch,

    2008-01-01

    boredomresearch exhibited the Real Snail Mail installation in Process as Paradigm Exhibition, LABoral Centro de Arte y Creacion, Gijon Spain (23rd April - 30th Aug 2010). A group exhibition which is showing artwork that is continually evolving and in a state of flux. the exhibition includes artworks by Ralf Baecker, Gregory Chatonsky, Peter Flemming, Roman Kirschner, C.E.B. Reas, Antoine Schmitt and Ralf Schreiber. Vicky Isley & Paul Smith (aka boredomresearch) in 2009 developed an installati...

  14. Real energy cost

    International Nuclear Information System (INIS)

    Vinogradova, I.

    1992-01-01

    Different methods of calculating the real power cost in the USA taking account of damage brought to the environment, public health expenses etc., are considered. Application of complex methods allowing one to directly determine the costs linked with ecology has shown that the most expensive power is generated at the new NPPs and thermal plants using coal. Activities on power saving and increasing the capacity of the existing hydroelectrotechnical equipment are considered to be the most effective from the viewpoint of expenses

  15. Real Time Processing

    CERN Multimedia

    CERN. Geneva; ANDERSON, Dustin James; DOGLIONI, Caterina

    2015-01-01

    The LHC provides experiments with an unprecedented amount of data. Experimental collaborations need to meet storage and computing requirements for the analysis of this data: this is often a limiting factor in the physics program that would be achievable if the whole dataset could be analysed. In this talk, I will describe the strategies adopted by the LHCb, CMS and ATLAS collaborations to overcome these limitations and make the most of LHC data: data parking, data scouting, and real-time analysis.

  16. Real and complex analysis

    CERN Document Server

    Apelian, Christopher; Taft, Earl; Nashed, Zuhair

    2009-01-01

    The Spaces R, Rk, and CThe Real Numbers RThe Real Spaces RkThe Complex Numbers CPoint-Set Topology Bounded SetsClassification of Points Open and Closed SetsNested Intervals and the Bolzano-Weierstrass Theorem Compactness and Connectedness Limits and Convergence Definitions and First Properties Convergence Results for SequencesTopological Results for Sequences Properties of Infinite SeriesManipulations of Series in RFunctions: Definitions and Limits DefinitionsFunctions as MappingsSome Elementary Complex FunctionsLimits of FunctionsFunctions: Continuity and Convergence Continuity Uniform Continuity Sequences and Series of FunctionsThe DerivativeThe Derivative for f: D1 → RThe Derivative for f: Dk → RThe Derivative for f: Dk → RpThe Derivative for f: D → CThe Inverse and Implicit Function TheoremsReal IntegrationThe Integral of f: [a, b] → RProperties of the Riemann Integral Further Development of Integration TheoryVector-Valued and Line IntegralsComplex IntegrationIntroduction to Complex Integrals Fu...

  17. REAL ESTATE PURCHASE AGREEMENTS

    Directory of Open Access Journals (Sweden)

    Bujorel FLOREA

    2016-12-01

    Full Text Available The study presented herein represents a field with good present and future perspectives, especially because real estate property is not under the incidence of a single normative act regarding the sale-purchase agreement of such goods, and given the fact that there are specific legal provisions with respect to various real estate categories and the localization of such property. The article deals with the sale-purchase agreement of various real estate categories, such as fields, buildings, the correspondent lots, urban area, farm, and forests fields, focusing on some particularities. A special care is attributed to examining the applicable laws with regard to the purchase agreements of field lands, the special conditions to be taken into account, the persons that may act as buyers, including foreigners, those without citizenship, and legal persons of a nationality other than Romanian. Finally, a special concern is given to the formalities required for legally exerting the pre-emptive right and the applicable sanctions in that respect.

  18. The real butterfly effect

    International Nuclear Information System (INIS)

    Palmer, T N; Döring, A; Seregin, G

    2014-01-01

    Historical evidence is reviewed to show that what Ed Lorenz meant by the iconic phrase ‘the butterfly effect’ is not at all captured by the notion of sensitive dependence on initial conditions in low-order chaos. Rather, as presented in his 1969 Tellus paper, Lorenz intended the phrase to describe the existence of an absolute finite-time predicability barrier in certain multi-scale fluid systems, implying a breakdown of continuous dependence on initial conditions for large enough forecast lead times. To distinguish from ‘mere’ sensitive dependence, the effect discussed in Lorenz's Tellus paper is referred to as ‘the real butterfly effect’. Theoretical evidence for such a predictability barrier in a fluid described by the three-dimensional Navier–Stokes equations is discussed. Whilst it is still an open question whether the Navier–Stokes equation has this property, evidence from both idealized atmospheric simulators and analysis of operational weather forecasts suggests that the real butterfly effect exists in an asymptotic sense, i.e. for initial-time atmospheric perturbations that are small in scale and amplitude compared with (weather) scales of interest, but still large in scale and amplitude compared with variability in the viscous subrange. Despite this, the real butterfly effect is an intermittent phenomenon in the atmosphere, and its presence can be signalled a priori, and hence mitigated, by ensemble forecast methods. (invited article)

  19. Several real variables

    CERN Document Server

    Kantorovitz, Shmuel

    2016-01-01

    This undergraduate textbook is based on lectures given by the author on the differential and integral calculus of functions of several real variables. The book has a modern approach and includes topics such as: •The p-norms on vector space and their equivalence •The Weierstrass and Stone-Weierstrass approximation theorems •The differential as a linear functional; Jacobians, Hessians, and Taylor's theorem in several variables •The Implicit Function Theorem for a system of equations, proved via Banach’s Fixed Point Theorem •Applications to Ordinary Differential Equations •Line integrals and an introduction to surface integrals This book features numerous examples, detailed proofs, as well as exercises at the end of sections. Many of the exercises have detailed solutions, making the book suitable for self-study. Several Real Variables will be useful for undergraduate students in mathematics who have completed first courses in linear algebra and analysis of one real variable.

  20. Development of Quantitative Real-Time PCR Assays for Detection and Quantification of Surrogate Biological Warfare Agents in Building Debris and Leachate▿

    Science.gov (United States)

    Saikaly, Pascal E.; Barlaz, Morton A.; de los Reyes, Francis L.

    2007-01-01

    Evaluation of the fate and transport of biological warfare (BW) agents in landfills requires the development of specific and sensitive detection assays. The objective of the current study was to develop and validate SYBR green quantitative real-time PCR (Q-PCR) assays for the specific detection and quantification of surrogate BW agents in synthetic building debris (SBD) and leachate. Bacillus atrophaeus (vegetative cells and spores) and Serratia marcescens were used as surrogates for Bacillus anthracis (anthrax) and Yersinia pestis (plague), respectively. The targets for SYBR green Q-PCR assays were the 16S-23S rRNA intergenic transcribed spacer (ITS) region and recA gene for B. atrophaeus and the gyrB, wzm, and recA genes for S. marcescens. All assays showed high specificity when tested against 5 ng of closely related Bacillus and Serratia nontarget DNA from 21 organisms. Several spore lysis methods that include a combination of one or more of freeze-thaw cycles, chemical lysis, hot detergent treatment, bead beat homogenization, and sonication were evaluated. All methods tested showed similar threshold cycle values. The limit of detection of the developed Q-PCR assays was determined using DNA extracted from a pure bacterial culture and DNA extracted from sterile water, leachate, and SBD samples spiked with increasing quantities of surrogates. The limit of detection for B. atrophaeus genomic DNA using the ITS and B. atrophaeus recA Q-PCR assays was 7.5 fg per PCR. The limits of detection of S. marcescens genomic DNA using the gyrB, wzm, and S. marcescens recA Q-PCR assays were 7.5 fg, 75 fg, and 7.5 fg per PCR, respectively. Quantification of B. atrophaeus vegetative cells and spores was linear (R2 > 0.98) over a 7-log-unit dynamic range down to 101 B. atrophaeus cells or spores. Quantification of S. marcescens (R2 > 0.98) was linear over a 6-log-unit dynamic range down to 102 S. marcescens cells. The developed Q-PCR assays are highly specific and sensitive and can

  1. Establishment of real time allele specific locked nucleic acid quantitative PCR for detection of HBV YIDD (ATT mutation and evaluation of its application.

    Directory of Open Access Journals (Sweden)

    Yongbin Zeng

    Full Text Available BACKGROUND: Long-term use of nucleos(tide analogues can increase risk of HBV drug-resistance mutations. The rtM204I (ATT coding for isoleucine is one of the most important resistance mutation sites. Establishing a simple, rapid, reliable and highly sensitive assay to detect the resistant mutants as early as possible is of great clinical significance. METHODS: Recombinant plasmids for HBV YMDD (tyrosine-methionine-aspartate-aspartate and YIDD (tyrosine-isoleucine-aspartate-aspartate were constructed by TA cloning. Real time allele specific locked nucleic acid quantitative PCR (RT-AS-LNA-qPCR with SYBR Green I was established by LNA-modified primers and evaluated with standard recombinant plasmids, clinical templates (the clinical wild type and mutant HBV DNA mixture and 102 serum samples from nucleos(tide analogues-experienced patients. The serum samples from a chronic hepatitis B (CHB patient firstly received LMV mono therapy and then switched to LMV + ADV combined therapy were also dynamically analyzed for 10 times. RESULTS: The linear range of the assay was between 1×10(9 copies/μl and 1 × 10(2 copies/μl. The low detection limit was 1 × 10(1 copies/μl. Sensitivity of the assay were 10(-6, 10(-4 and 10(-2 in the wild-type background of 1 × 10(9 copies/μl, 1 × 10(7 copies/μl and 1 × 10(5 copies/μl, respectively. The sensitivity of the assay in detection of clinical samples was 0.03%. The complete coincidence rate between RT-AS-LNA-qPCR and direct sequencing was 91.2% (93/102, partial coincidence rate was 8.8% (9/102, and no complete discordance was observed. The two assays showed a high concordance (Kappa = 0.676, P = 0.000. Minor variants can be detected 18 weeks earlier than the rebound of HBV DNA load and alanine aminotransferase level. CONCLUSIONS: A rapid, cost-effective, high sensitive, specific and reliable method of RT-AS-LNA-qPCR with SYBR Green I for early and absolute quantification of HBV YIDD (ATT coding for isoleucine

  2. Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp. cubense tropical race 4 in soil.

    Directory of Open Access Journals (Sweden)

    Xin Zhang

    Full Text Available Fusarium oxysporum f. sp. cubense (Foc, the causal agent of Fusarium wilt (Panama disease, is one of the most devastating diseases of banana (Musa spp.. The Foc tropical race 4 (TR4 is currently known as a major concern in global banana production. No effective resistance is known in Musa to Foc, and no effective measures for controlling Foc once banana plants have been infected in place. Early and accurate detection of Foc TR4 is essential to protect banana industry and guide banana planting. A real-time fluorescence loop-mediated isothermal amplification assay (RealAmp was developed for the rapid and quantitative detection of Foc TR4 in soil. The detection limit of the RealAmp assay was approximately 0.4 pg/µl plasmid DNA when mixed with extracted soil DNA or 10(3 spores/g of artificial infested soil, and no cross-reaction with other relative pathogens were observed. The RealAmp assay for quantifying genomic DNA of TR4 was confirmed by testing both artificially and naturally infested samples. Quantification of the soil-borne pathogen DNA of Foc TR4 in naturally infested samples was no significant difference compared to classic real-time PCR (P>0.05. Additionally, RealAmp assay was visual with an improved closed-tube visual detection system by adding SYBR Green I fluorescent dye to the inside of the lid prior to amplification, which avoided the inhibitory effects of the stain on DNA amplification and makes the assay more convenient in the field and could thus become a simple, rapid and effective technique that has potential as an alternative tool for the detection and monitoring of Foc TR4 in field, which would be a routine DNA-based testing service for the soil-borne pathogen in South China.

  3. Real three-dimensional biquadrics

    Energy Technology Data Exchange (ETDEWEB)

    Krasnov, Vyacheslav A [P.G. Demidov Yaroslavl State University, Yaroslavl (Russian Federation)

    2010-09-07

    We find the topological types of biquadrics (complete intersections of two real four-dimensional quadrics). The rigid isotopy classes of real three-dimensional biquadrics were described long ago: there are nine such classes. We find the correspondence between the topological types of real biquadrics and their rigid isotopy classes, and show that only two rigid isotopy classes have the same topological type. One of these classes consists of real GM-varieties and the other contains no GM-varieties. We also study the sets of real lines on real biquadrics.

  4. A specific endogenous reference for genetically modified common bean (Phaseolus vulgaris L.) DNA quantification by real-time PCR targeting lectin gene.

    Science.gov (United States)

    Venturelli, Gustavo L; Brod, Fábio C A; Rossi, Gabriela B; Zimmermann, Naíra F; Oliveira, Jaison P; Faria, Josias C; Arisi, Ana C M

    2014-11-01

    The Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new taxon-specific endogenous reference quantification assay was developed for Phaseolus vulgaris L. Three genes encoding common bean proteins (phaseolin, arcelin, and lectin) were selected as candidates for endogenous reference. Primers targeting these candidate genes were designed and the detection was evaluated using the SYBR Green chemistry. The assay targeting lectin gene showed higher specificity than the remaining assays, and a hydrolysis probe was then designed. This assay showed high specificity for 50 common bean samples from two gene pools, Andean and Mesoamerican. For GM common bean varieties, the results were similar to those obtained for non-GM isogenic varieties with PCR efficiency values ranging from 92 to 101 %. Moreover, this assay presented a limit of detection of ten haploid genome copies. The primers and probe developed in this work are suitable to detect and quantify either GM or non-GM common bean.

  5. Detection of five potentially periodontal pathogenic bacteria in peri-implant disease: A comparison of PCR and real-time PCR.

    Science.gov (United States)

    Schmalz, Gerhard; Tsigaras, Sandra; Rinke, Sven; Kottmann, Tanja; Haak, Rainer; Ziebolz, Dirk

    2016-07-01

    The aim of this study was to compare the microbial analysis methods of polymerase chain reaction (PCR) and real-time PCR (RT-PCR) in terms of detection of five selected potentially periodontal pathogenic bacteria in peri-implant disease. Therefore 45 samples of healthy, mucositis and peri-implantitis (n = 15 each) were assessed according to presence of the following bacteria using PCR (DNA-strip technology) and RT-PCR (fluorescent dye SYBR green-system): Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tanerella forsythia (Tf), and Fusobacterium nucleatum (Fn). There were no significant correlations between the bacterial and disease patterns, so the benefit of using microbiological tests for the diagnosis of peri-implant diseases is questionable. Correlations between the methods were highest for Tf (Kendall's Tau: 0.65, Spearman: 0.78), Fn (0.49, 0.61) and Td (0.49, 0.59). For Aa (0.38, 0.42) and Pg (0.04, 0.04), lower correlation values were detected. Accordingly, conventional semi-quantitative PCR seems to be sufficient for analyzing potentially periodontal pathogenic bacterial species. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Development and application of a quantitative real-time PCR assay for rapid detection of the multifaceted yeast Kazachstania servazzii in food.

    Science.gov (United States)

    Spanoghe, Martin; Godoy Jara, Mario; Rivière, John; Lanterbecq, Deborah; Gadenne, Martine; Marique, Thierry

    2017-04-01

    The beneficial contributions of Kazachstania servazzii are well-established in various food processes. This yeast also contributes in the spoilage of finished packaged food due to abundant gas production. In particular, an occurrence of K. servazzii was recently positively correlated with the formation of severe package swelling of some prepared fresh pizzas. To circumscribe this concern, a quantitative SYBR green real-time PCR assay based on a newly designed specific primer pair targeting the ribosomal ITS1-5.8S-ITS2 region of K. servazzii was developed. The quantification was enabled using a standard curve created from serially diluted plasmids containing the target sequence of the K. servazzii strain. A validation of the assay was achieved by enumeration of K. servazzii DNA copies from artificially infected culture broths containing non-contaminated pizza substrates. The newly developed method was then tested on total DNA extracted from packaged fresh pizzas, in which certain lots were swollen and thus suspected of containing K. servazzii. This study highlights that this newly developed quantitative assay is not only sufficiently sensitive, specific and reliable to be functionally used in food control as a routine method of detection, but also promising in specific studies that seek to further characterize the dynamic of this yeast in some increasingly popular food processes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Ovation Prime Real-Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Ovation Prime Real-Time (OPRT) product is a real-time forecast and nowcast model of auroral power and is an operational implementation of the work by Newell et...

  8. Real Time Text Analysis

    Science.gov (United States)

    Senthilkumar, K.; Ruchika Mehra Vijayan, E.

    2017-11-01

    This paper aims to illustrate real time analysis of large scale data. For practical implementation we are performing sentiment analysis on live Twitter feeds for each individual tweet. To analyze sentiments we will train our data model on sentiWordNet, a polarity assigned wordNet sample by Princeton University. Our main objective will be to efficiency analyze large scale data on the fly using distributed computation. Apache Spark and Apache Hadoop eco system is used as distributed computation platform with Java as development language

  9. Real time Faraday spectrometer

    International Nuclear Information System (INIS)

    Smith, T.E.; Struve, K.W.; Colella, N.J.

    1991-01-01

    This patent describes an invention which uses a dipole magnet to bend the path of a charged particle beam. As the deflected particles exit the magnet, they are spatially dispersed in the bend-plane of the magnet according to their respective momenta and pass to a plurality of chambers having Faraday probes positioned therein. Both the current and energy distribution of the particles is then determined by the non-intersecting Faraday probes located along the chambers. The Faraday probes are magnetically isolated from each other by thin metal walls of the chambers, effectively providing real time current-versus-energy particle measurements

  10. Real four-dimensional biquadrics

    Energy Technology Data Exchange (ETDEWEB)

    Krasnov, Vyacheslav A [P.G. Demidov Yaroslavl State University, Yaroslavl (Russian Federation)

    2011-04-30

    We consider intersections of two real five-dimensional quadrics, which are referred to for brevity as real four-dimensional biquadrics. Their rigid isotopy classes were described long ago: there are 16 such classes. We prove that the rigid isotopy class of a non-singular real four-dimensional biquadric is uniquely determined by the topological type of its real part. To do this, we calculate the dimensions of the cohomology spaces of the real part of a four-dimensional biquadric.

  11. Differentiation of real functions

    CERN Document Server

    Bruckner, Andrew

    1994-01-01

    Topics related to the differentiation of real functions have received considerable attention during the last few decades. This book provides an efficient account of the present state of the subject. Bruckner addresses in detail the problems that arise when dealing with the class \\Delta ' of derivatives, a class that is difficult to handle for a number of reasons. Several generalized forms of differentiation have assumed importance in the solution of various problems. Some generalized derivatives are excellent substitutes for the ordinary derivative when the latter is not known to exist; others are not. Bruckner studies generalized derivatives and indicates "geometric" conditions that determine whether or not a generalized derivative will be a good substitute for the ordinary derivative. There are a number of classes of functions closely linked to differentiation theory, and these are examined in some detail. The book unifies many important results from the literature as well as some results not previously pub...

  12. Real time production optimization

    Energy Technology Data Exchange (ETDEWEB)

    Saputelli, Luigi; Otavio, Joao; Araujo, Turiassu; Escorcia, Alvaro [Halliburton, Houston, TX (United States). Landmark Division

    2004-07-01

    Production optimization encompasses various activities of measuring, analyzing, modeling, prioritizing and implementing actions to enhance productivity of a field. We present a state-of-the-art framework for optimizing production on a continuous basis as new sensor data is acquired in real time. Permanently acquired data is modeled and analyzed in order to create predictive models. A model based control strategy is used to regulate well and field instrumentation. The optimum field operating point, which changes with time, satisfies the maximum economic return. This work is a starting point for further development in automatic, intelligent reservoir technologies which get the most out of the abilities of permanent, instrumented wells and remotely activated downhole completions. The strategy, tested with history-matched data from a compartmentalised giant field, proved to reduce operating costs while increasing oil recovery by 27% in this field. (author)

  13. Real algebraic geometry

    CERN Document Server

    Bochnak, Jacek; Roy, Marie-Françoise

    1998-01-01

    This book is a systematic treatment of real algebraic geometry, a subject that has strong interrelation with other areas of mathematics: singularity theory, differential topology, quadratic forms, commutative algebra, model theory, complexity theory etc. The careful and clearly written account covers both basic concepts and up-to-date research topics. It may be used as text for a graduate course. The present edition is a substantially revised and expanded English version of the book "Géometrie algébrique réelle" originally published in French, in 1987, as Volume 12 of ERGEBNISSE. Since the publication of the French version the theory has made advances in several directions. Many of these are included in this English version. Thus the English book may be regarded as a completely new treatment of the subject.

  14. Real time urbanism

    Directory of Open Access Journals (Sweden)

    Ana Ruiz Varona

    2012-12-01

    Full Text Available Nowadays, given the technological revolution of the society of information, the administrative management of the cities faces a new problem not as related to the projection of the urban space as to the capacity of controlling and measuring the process of direct and centralized production of the cities by part of some non-homogeneous social multitudes, in a hyper-accelerated time towards instantaneity. Against libertarian apologies of the new “participative urbanisms”, the article puts forward a discourse that shows the lost associated to the new problem of temporal instantaneity. In this regard we claim new process of mediation that allow administrations and urbanist monitoring the production of the city. To that end, a previous and necessary step will be the redefinition of the role of a new real time urbanist.

  15. Real-world outcomes

    DEFF Research Database (Denmark)

    Askehave, Inger; Holmgreen, Lise-Lotte

    2012-01-01

    analyses during one specific meeting. An important element of the latter is to assess whether the discussion of our results gives rise to respondents’ reflection, resistance, acceptance, rejection, or indifference, and whether this leads to changes in respondents’ views on culture and organizational......When working with discourse-based research in professional settings, one question invariably arises: How do we, as researchers, ensure that our findings are taken up by the professionals with whom analyses are carried out, leading to change in their social practices? The answer is that quite often...... of research findings may take place to ensure the creation of real-world outcomes for practitioners. The discussion will be centered around the interview study of the discursive constructions of culture in a Danish cross-border company and the possible implications of this for organizational collaboration...

  16. Threshold microsclerotial inoculum for cotton verticillium wilt determined through wet-sieving and real-time quantitative PCR.

    Science.gov (United States)

    Wei, Feng; Fan, Rong; Dong, Haitao; Shang, Wenjing; Xu, Xiangming; Zhu, Heqin; Yang, Jiarong; Hu, Xiaoping

    2015-02-01

    Quantification of Verticillium dahliae microsclerotia is an important component of wilt management on a range of crops. Estimation of microsclerotia by dry or wet sieving and plating of soil samples on semiselective medium is a commonly used technique but this method is resource-intensive. We developed a new molecular quantification method based on Synergy Brands (SYBR) Green real-time quantitative polymerase chain reaction of wet-sieving samples (wet-sieving qPCR). This method can detect V. dahliae microsclerotia as low as 0.5 CFU g(-1) of soil. There was a high correlation (r=0.98) between the estimates of conventional plating analysis and the new wet-sieving qPCR method for 40 soil samples. To estimate the inoculum threshold for cotton wilt, >400 soil samples were taken from the rhizosphere of individual plants with or without visual wilt symptoms in experimental and commercial cotton fields at the boll-forming stage. Wilt inoculum was estimated using the wet-sieving qPCR method and related to wilt development. The estimated inoculum threshold varied with cultivar, ranging from 4.0 and 7.0 CFU g(-1) of soil for susceptible and resistant cultivars, respectively. In addition, there was an overall relationship of wilt incidence with inoculum density across 31 commercial fields where a single composite soil sample was taken at each field, with an estimated inoculum threshold of 11 CFU g(-1) of soil. These results suggest that wilt risk can be predicted from the estimated soil inoculum density using the new wet-sieving qPCR method. We recommend the use of 4.0 and 7.0 CFU g(-1) as an inoculum threshold on susceptible and resistant cultivars, respectively, in practical risk prediction schemes.

  17. Identificação de três espécies de Candida por PCR em tempo real

    Directory of Open Access Journals (Sweden)

    Alessandra Koehler

    2016-06-01

    Full Text Available Infecções causadas por leveduras do gênero Candida são muito prevalentes. A correta identificação da espécie é uma etapa fundamental para a escolha do tratamento adequado, mas os métodos tradicionais apresentam limitações. Assim, o objetivo do presente trabalho foi padronizar uma reação de PCR em tempo real (qPCR para identificação de Candida albicans, C. glabrata e C. parapsilosis, que é uma técnica mais rápida para o diagnóstico. As amostras utilizadas consistiram em DNA extraído de cepas padrão. Realizou-se uma qPCR com SybrGreen® e diluições seriadas das amostras, com as mesmas condições do trabalho em que foram descritos os primers utilizados. Nesse ensaio os resultados foram satisfatórios apenas para C. parapsilosis, pois em C. albicans e C. glabrata, apesar de ocorrer a amplificação da sequência alvo, houve formação de produtos inespecíficos nas reações. Foi feita então uma análise in silico dos primers, que sugeriu não haver problemas significativos em sua construção, contudo encontrou-se alguns problemas de especificidade para C. parapsilosis e C. glabrata. Conclui-se que são necessários estudos posteriores para desenvolvimento e validação de novos primers que mostrem maior sensibilidade e especificidade para as espécies patogênicas.

  18. UML for real design of embedded real-time systems

    CERN Document Server

    Martin, Grant; Selic, Bran

    2003-01-01

    Models, Software Models and UML.- UML for Real-Time.- Structural Modeling with UML 2.0.- Message Sequence Charts.- UML and Platform-based Design.- UML for Hardware and Software Object Modeling.- Fine Grained Patterns for Real-Time Systems.- Architectural Patterns for Real-Time Systems.- Modeling Quality of Service with UML.- Modeling Metric Time.- Performance Analysis with UML.- Schedulability Analysis with UML.- Automotive UML.- Specifying Telecommunications Systems with UML.- Leveraging UML to Deliver Correct Telecom Applications.- Software Performance Engineering.

  19. Real-estate lakes

    Science.gov (United States)

    Rickert, David A.; Spieker, Andrew Maute

    1971-01-01

    Since the dawn of civilization waterfront land has been an irresistible attraction to man. Throughout history he has sought out locations fronting on oceans, rivers, and lakes. Originally sought for proximity .to water supply and transportation, such locations are now sought more for their esthetic qualities and for recreation. Usable natural waterfront property is limited, however, and the more desirable sites in many of our urban areas have already been taken. The lack of available waterfront sites has led to the creation of many artificial bodies of water. The rapid suburbanization that has characterized urban growth in America since the end of World War II, together with increasing affluence and le-isure time, has created a ready market for waterfront property. Accordingly, lake-centered subdivisions and developments dot the suburban landscape in many of our major urban areas. Literally thousands of lakes surrounded by homes have materialized during this period of rapid growth. Recently, several "new town" communities have been planned around this lake-centered concept. A lake can be either an asset or a liaoility to a community. A clean, clear, attractively landscaped lake is a definite asset, whereas a weed-choked, foul-smelling mudhole is a distinct liability. The urban environment poses both problems and imaginative opportunities in the development of lakes. Creation of a lake causes changes in all aspects of the environment. Hydrologic systems and ecological patterns are usually most severely altered. The developer should be aware of the potential changes; it is not sufficient merely to build a dam across a stream or to dig a hole in the ground. Development of Gl a successful lake requires careful planning for site selection and design, followed by thorough and cc ntinual management. The purpose of this report is to describe the characteristics of real-estate lakes, to pinpoint potential pmblems, and to suggest possible planning and management guidelines

  20. An introduction to real analysis

    CERN Document Server

    Ball, Derek G

    1973-01-01

    An Introduction to Real Analysis presents the concepts of real analysis and highlights the problems which necessitate the introduction of these concepts. Topics range from sets, relations, and functions to numbers, sequences, series, derivatives, and the Riemann integral. This volume begins with an introduction to some of the problems which are met in the use of numbers for measuring, and which provide motivation for the creation of real analysis. Attention then turns to real numbers that are built up from natural numbers, with emphasis on integers, rationals, and irrationals. The chapters tha

  1. Species determination of fowl adenoviruses based on the 52K gene region.

    Science.gov (United States)

    Günes, Ayse; Marek, Ana; Hess, Michael

    2013-06-01

    In the present study, the classification of fowl adenoviruses (FAdVs) based on a part of the 52K gene region was described. A total of 44 FAdV field samples from different countries and sources were detected using a recently developed SYBR Green-based real-time PCR. Amplified products were sequenced, and phylogenetic analyses were conducted on the basis of the 116-bp region. For comparison, the already published sequences of the 52K gene region of aviadenoviruses were used in the analyses. The phylogenetic analysis allowed the grouping of the FAdVs into the established five different FAdV species: Fowl adenovirus A to Fowl adenovirus E. The existence of the species was supported by high bootstrap values (> 70%). This method provides the advantages of quantitation and high sensitivity for FAdV detection in combination with species assignment.

  2. Investigations on the frequency of norovirus contamination of ready-to-eat food items in Istanbul, Turkey, by using real-time reverse transcription PCR.

    Science.gov (United States)

    Yilmaz, Aysun; Bostan, Kamil; Altan, Eda; Muratoglu, Karlo; Turan, Nuri; Tan, Derya; Helps, Christopher; Yilmaz, Huseyin

    2011-05-01

    Investigation of norovirus (NoV) contamination of food items is important because many outbreaks occur after consumption of contaminated shellfish, vegetables, fruits, and water. The frequency of NoV contamination in food items has not previously been investigated in Turkey. The aim of this study was to investigate the frequency of human NoV genogroups (G) I and II in ready-to-eat tomatoes, parsley, green onion, lettuce, mixed salads, and cracked wheat balls. RNA was extracted with the RNeasy Mini Kit, and a real-time reverse transcription (RT) PCR assay was performed using primers specific for NoV GI and GII. Among the 525 samples analyzed, NoV GII was detected in 1 green onion sample and 1 tomato sample by both SYBR Green and TaqMan real-time RT-PCR assays; no GI virus was detected. The Enterobactericaeae and Escherichia coli levels in the NoV-positive green onion were 6.56 and 1.28 log CFU/g, and those in the tomato were 5.55 and 1.30 log CFU/g, respectively. No significant difference in the bacterial levels was found between the NoV-positive and NoV-negative samples. This study is the first in which NoV GII was found in ready-to-eat food collected from Istanbul, Turkey; thus, these foods may be considered a risk to human health. Epidemiological studies and measures to prevent NoV infection should be considered.

  3. Validation of Reference Genes for Relative Quantitative Gene Expression Studies in Cassava (Manihot esculenta Crantz) by Using Quantitative Real-Time PCR.

    Science.gov (United States)

    Hu, Meizhen; Hu, Wenbin; Xia, Zhiqiang; Zhou, Xincheng; Wang, Wenquan

    2016-01-01

    Reverse transcription quantitative real-time polymerase chain reaction (real-time PCR, also referred to as quantitative RT-PCR or RT-qPCR) is a highly sensitive and high-throughput method used to study gene expression. Despite the numerous advantages of RT-qPCR, its accuracy is strongly influenced by the stability of internal reference genes used for normalizations. To date, few studies on the identification of reference genes have been performed on cassava (Manihot esculenta Crantz). Therefore, we selected 26 candidate reference genes mainly via the three following channels: reference genes used in previous studies on cassava, the orthologs of the most stable Arabidopsis genes, and the sequences obtained from 32 cassava transcriptome sequence data. Then, we employed ABI 7900 HT and SYBR Green PCR mix to assess the expression of these genes in 21 materials obtained from various cassava samples under different developmental and environmental conditions. The stability of gene expression was analyzed using two statistical algorithms, namely geNorm and NormFinder. geNorm software suggests the combination of cassava4.1_017977 and cassava4.1_006391 as sufficient reference genes for major cassava samples, the union of cassava4.1_014335 and cassava4.1_006884 as best choice for drought stressed samples, and the association of cassava4.1_012496 and cassava4.1_006391 as optimal choice for normally grown samples. NormFinder software recommends cassava4.1_006884 or cassava4.1_006776 as superior reference for qPCR analysis of different materials and organs of drought stressed or normally grown cassava, respectively. Results provide an important resource for cassava reference genes under specific conditions. The limitations of these findings were also discussed. Furthermore, we suggested some strategies that may be used to select candidate reference genes.

  4. The real new economy.

    Science.gov (United States)

    Farrell, Diana

    2003-10-01

    During the soar-and-swoon days of the late 1990s, many people believed that information technology, and the Internet in particular, were "changing everything" in business. A fundamental change did happen in the 1990s, but it was less about technology than about competition. Under director Diana Farrell, the McKinsey Global Institute has conducted an extensive study of productivity and its connection to corporate IT spending and use during that period. The study revealed that information technology is important--but not central--to the fate of industries and individual companies. So if information technology was not the primary factor in the productivity surge, what was? The study points to competition and innovation. In those industries that saw increases in competitive intensity, managers were forced to innovate aggressively to protect their revenues and profits. Those innovations--in products, business practices, and technology--led to the gains in productivity. In fact, a critical dynamic of the new economy--the real new economy--is the virtuous cycle of competition, innovation, and productivity growth. Managers can innovate in many ways, but during the 1990s, information technology was a particularly powerful tool, for three reasons: First, IT enabled the development of attractive new products and efficient new business processes. Second, it facilitated the rapid industrywide diffusion of innovations. And third, it exhibited strong scale economies--its benefits multiplied rapidly as its use expanded. This article reveals surprising data on how various industries in the United States and Europe were affected by competition, innovation, and information technology in the 1990s and offers insights about how managers can get more from their IT investments.

  5. Real Life and the Classroom

    Science.gov (United States)

    Van Praagh, Shauna

    2013-01-01

    The choices teachers make about both form and content in designing their courses shape students' perceptions of the connection between "real life" and the "classroom." The choice to begin a first-year law course with stories told by residential school survivors provides an example of how to bring "real life" into the…

  6. Analysing UK real estate market forecast disagreement

    OpenAIRE

    McAllister, Patrick; Newell, G.; Matysiak, George

    2005-01-01

    Given the significance of forecasting in real estate investment decisions, this paper investigates forecast uncertainty and disagreement in real estate market forecasts. Using the Investment Property Forum (IPF) quarterly survey amongst UK independent real estate forecasters, these real estate forecasts are compared with actual real estate performance to assess a number of real estate forecasting issues in the UK over 1999-2004, including real estate forecast error, bias and consensus. The re...

  7. Economic Effects Real Estate Tax

    Directory of Open Access Journals (Sweden)

    Tadić Milan

    2016-06-01

    Full Text Available The real estate tax is usually a fiscal instrument which performs the property tax. When it comes to real property or immovable this term include: apartments, houses, land, cottages, excess housing landscape and more. The real estate tax as a form of the fiscal charges ownership or use of certain forms of real estate, and the revenue from this tax is levied on the area where the property is located regardless of the place of residence of its owner. The tax base for the calculation of this tax usually consists of the market, estimated or annuity value of certain real estate. This form of taxation in the Republic of Serbian applies from 1.1.2012., and its introduction has been replaced by former property taxes. The differences between the two concepts mentioned taxes are numerous and significant. Among the more important are: subject to taxation under the new concept of the real estate rather than law, a taxpayer is any property owner rather than the holder of rights to immovable property tax base is the market value of real estate which is replaced by the payment of taxes per square meter of usable area, the rate of property tax is determined local government, which can not be lower than 0.05% of the estimated value of the real estate nor higher than 0.5% of the appraised value of real estate. The last change, ie. The new law on Property Tax from 5.11.2015. was determined by the tax rate to 20%. The fact that local governments each of them determines the tax rate on real estate which range from high to low rates of multiple, makes this tax is progressive. Progression is particularly expressed in the distinction applied tax rates of developed and undeveloped municipalities, where we have a case that less developed tolerate a higher tax burden, which leads to negative economic effects. However, real estate tax has its own economic and social characteristics which must be aligned with the objectives of tax policy. This means that the real estate tax

  8. Strange functions in real analysis

    CERN Document Server

    Kharazishvili, AB

    2005-01-01

    Weierstrass and Blancmange nowhere differentiable functions, Lebesgue integrable functions with everywhere divergent Fourier series, and various nonintegrable Lebesgue measurable functions. While dubbed strange or "pathological," these functions are ubiquitous throughout mathematics and play an important role in analysis, not only as counterexamples of seemingly true and natural statements, but also to stimulate and inspire the further development of real analysis.Strange Functions in Real Analysis explores a number of important examples and constructions of pathological functions. After introducing the basic concepts, the author begins with Cantor and Peano-type functions, then moves to functions whose constructions require essentially noneffective methods. These include functions without the Baire property, functions associated with a Hamel basis of the real line, and Sierpinski-Zygmund functions that are discontinuous on each subset of the real line having the cardinality continuum. Finally, he considers e...

  9. Real-Time Parameter Identification

    Data.gov (United States)

    National Aeronautics and Space Administration — Armstrong researchers have implemented in the control room a technique for estimating in real time the aerodynamic parameters that describe the stability and control...

  10. From real to complex analysis

    CERN Document Server

    Dyer, R H

    2014-01-01

    The purpose of this book is to provide an integrated course in real and complex analysis for those who have already taken a preliminary course in real analysis. It particularly emphasises the interplay between analysis and topology. Beginning with the theory of the Riemann integral (and its improper extension) on the real line, the fundamentals of metric spaces are then developed, with special attention being paid to connectedness, simple connectedness and various forms of homotopy. The final chapter develops the theory of complex analysis, in which emphasis is placed on the argument, the winding number, and a general (homology) version of Cauchy's theorem which is proved using the approach due to Dixon. Special features are the inclusion of proofs of Montel's theorem, the Riemann mapping theorem and the Jordan curve theorem that arise naturally from the earlier development. Extensive exercises are included in each of the chapters, detailed solutions of the majority of which are given at the end. From Real to...

  11. Real and Hybrid Atomic Orbitals.

    Science.gov (United States)

    Cook, D. B.; Fowler, P. W.

    1981-01-01

    Demonstrates that the Schrodinger equation for the hydrogenlike atom separates in both spheroconal and prolate spheroidal coordinates and that these separations provide a sound theoretical basis for the real and hybrid atomic orbitals. (Author/SK)

  12. Real analysis with economic applications

    CERN Document Server

    Ok, Efe A

    2011-01-01

    There are many mathematics textbooks on real analysis, but they focus on topics not readily helpful for studying economic theory or they are inaccessible to most graduate students of economics. Real Analysis with Economic Applications aims to fill this gap by providing an ideal textbook and reference on real analysis tailored specifically to the concerns of such students. The emphasis throughout is on topics directly relevant to economic theory. In addition to addressing the usual topics of real analysis, this book discusses the elements of order theory, convex analysis, optimization, correspondences, linear and nonlinear functional analysis, fixed-point theory, dynamic programming, and calculus of variations. Efe Ok complements the mathematical development with applications that provide concise introductions to various topics from economic theory, including individual decision theory and games, welfare economics, information theory, general equilibrium and finance, and intertemporal economics. Moreover, a...

  13. Determining Real Estate Licensee Income

    OpenAIRE

    G. Stacy Sirmans; Philip G. Swicegood

    2000-01-01

    This article examines the determinants of real estate licensee income using a 1997 survey of Texas real estate licensees. The factors having a positive effect on licensee income include: (1) number of hours worked; (2) work experience; (3) being a male; (4) using computer technology; (5) being involved in more transactions; (6) holding professional designations; (7) being associated with a larger firm; and (8) having access to personal assistants. Variables that negatively affect income inclu...

  14. Real-time vision systems

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, R.; Hernandez, J.E.; Lu, Shin-yee [Lawrence Livermore National Lab., CA (United States)

    1994-11-15

    Many industrial and defence applications require an ability to make instantaneous decisions based on sensor input of a time varying process. Such systems are referred to as `real-time systems` because they process and act on data as it occurs in time. When a vision sensor is used in a real-time system, the processing demands can be quite substantial, with typical data rates of 10-20 million samples per second. A real-time Machine Vision Laboratory (MVL) was established in FY94 to extend our years of experience in developing computer vision algorithms to include the development and implementation of real-time vision systems. The laboratory is equipped with a variety of hardware components, including Datacube image acquisition and processing boards, a Sun workstation, and several different types of CCD cameras, including monochrome and color area cameras and analog and digital line-scan cameras. The equipment is reconfigurable for prototyping different applications. This facility has been used to support several programs at LLNL, including O Division`s Peacemaker and Deadeye Projects as well as the CRADA with the U.S. Textile Industry, CAFE (Computer Aided Fabric Inspection). To date, we have successfully demonstrated several real-time applications: bullet tracking, stereo tracking and ranging, and web inspection. This work has been documented in the ongoing development of a real-time software library.

  15. 15 CFR 806.8 - Real estate.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 3 2010-01-01 2010-01-01 false Real estate. 806.8 Section 806.8... ECONOMIC ANALYSIS, DEPARTMENT OF COMMERCE DIRECT INVESTMENT SURVEYS § 806.8 Real estate. Residential real..., is considered real estate held for personal use. Ownership of residential real estate by a...

  16. Real Estate Agent Commission Disputes

    Directory of Open Access Journals (Sweden)

    Anida Duarte

    2015-06-01

    Full Text Available The purpose of this study was to examine the relationship among Procuring Cause Law, real estate agent years of experience, and real estate commission disputes. A pilot survey was conducted in the southwestern United States among real estate agents, realtor(s, and brokers. After testing the hypothesis, the decision was made to fail to reject the hypothesis and conclude that real estate agent experience and not Procuring Cause Law produced favorable outcomes in disputes. As a result, the following recommendations were made: (a Agency seller and buyer’s agreements should be used in each transaction to avoid disputes, (b proper expectations and guidelines should be reviewed prior to starting any real estate transaction, (c a checklist may assist in the assurance that all valuable information is reviewed, (d agents could benefit from fully understanding Procuring Cause Law and sharing this information with their clients, (e state and national regulatory requirements of the law could be modified for easier understanding and use, and (f consumers who willfully violate the law could be subject to monetary penalties.

  17. Real-time PCR detection of 16S rRNA genes speeds most-probable-number enumeration of foodborne Listeria monocytogenes.

    Science.gov (United States)

    De Martinis, Elaine Cristina Pereira; Duvall, Robert E; Hitchins, Anthony D

    2007-07-01

    Quantifying foodborne pathogens at concentrations of 0.1 to 1,000 CFU/g of food generally involves most-probable-number (MPN) enumeration, which takes at least 4 days. A real-time PCR assay (RTi-PCR) was developed to accelerate MPN enumeration of foodborne Listeria monocytogenes. Foods were spiked from 70 to 110 CFU/g, and triplicate subportions from 0.0001 to 1 g were selectively enriched for 48 h at 30 degrees C. For standard MPN enumeration, the enrichments were subcultured on Oxford agar (48 h at 35 degrees C) to isolate Listeria. For RTi-PCR MPN, the L. monocytogenes cells from the same enrichments were washed and resuspended in 2 ml of sterile water. DNA was extracted by boiling for 10 min. The DNA in the extract's supernatant was targeted with published oligonucleotide primers for amplifying an Lmo-specific sequence of 16S rRNA genes. Amplification was continuously monitored with SYBR Green. The resulting amplicon was characterized by its melting temperature. The L. monocytogenes specificity of the primers was confirmed by testing L. monocytogenes (15 strains), Listeria innocua (11 strains), and Listeria welshimeri, Listeria seeligeri, Listeria ivanovii, and Listeria grayi (1 strain each). Quantitatively spiked milk, lettuce, smoked salmon, Brie cheese, ice cream, pork pâté, salami, ready-to-eat shrimp, raw ground beef, and fresh soft cheese were enumerated by both the standard and the PCR MPN method. The paired results from the two MPN methods agreed well, except for the fresh cheese. For some foods, l-g samples required a decimal dilution for a positive test result, suggesting concentration-dependent food ingredient interference with the RTi-PCR. This RTi-PCR method reduced the time necessary for the MPN enumeration of foodborne L. monocytogenes from 4 to 2 days.

  18. A Real Time PCR Platform for the Simultaneous Quantification of Total and Extrachromosomal HIV DNA Forms in Blood of HIV-1 Infected Patients

    Science.gov (United States)

    Canovari, Benedetta; Scotti, Maddalena; Acetoso, Marcello; Valentini, Massimo; Petrelli, Enzo; Magnani, Mauro

    2014-01-01

    Background The quantitative measurement of various HIV-1 DNA forms including total, unintegrated and integrated provirus play an increasingly important role in HIV-1 infection monitoring and treatment-related research. We report the development and validation of a SYBR Green real time PCR (TotUFsys platform) for the simultaneous quantification of total and extrachromosomal HIV-1 DNA forms in patients. This innovative technique makes it possible to obtain both measurements in a single PCR run starting from frozen blood employing the same primers and standard curve. Moreover, due to identical amplification efficiency, it allows indirect estimation of integrated level. To specifically detect 2-LTR a qPCR method was also developed. Methodology/Findings Primers used for total HIV-1 DNA quantification spanning a highly conserved region were selected and found to detect all HIV-1 clades of group M and the unintegrated forms of the same. A total of 195 samples from HIV-1 patients in a wide range of clinical conditions were analyzed with a 100% success rate, even in patients with suppressed plasma viremia, regardless of CD4+ or therapy. No significant correlation was observed between the two current prognostic markers, CD4+ and plasma viremia, while a moderate or high inverse correlation was found between CD4+ and total HIV DNA, with strong values for unintegrated HIV DNA. Conclusions/Significance Taken together, the results support the use of HIV DNA as another tool, in addition to traditional assays, which can be used to estimate the state of viral infection, the risk of disease progression and to monitor the effects of ART. The TotUFsys platform allowed us to obtain a final result, expressed as the total and unintegrated HIV DNA copy number per microgram of DNA or 104 CD4+, for 12 patients within two working days. PMID:25364909

  19. Complex analogues of real problems

    DEFF Research Database (Denmark)

    Esdahl-Schou, Rune

    This thesis will be a mix of different problems in number theory. As such it is split into two natural parts. The rst part focuses on normal numbers and construction of numbers that are normal to a given complex base. It is written in the style of a thorough and introductory paper on that subject...... and Velani, and inspired by this An gave a different proof which provides a stronger result. The conjecture is concerned with intersections of certain sets in the plane and are as such a real problem. We will consider a slightly different setup where the real plane is replaced by the complex plane. Using...... geometrical interpretations we construct sets with properties similar to the sets considered in the real case. We then formulate a conjecture which can be interpreted as a complex version of Schmidt's original conjecture. Finally we construct a variant of Schmidt's game, to show a partial result leading us...

  20. Designing Real Time Assistive Technologies

    DEFF Research Database (Denmark)

    Sonne, Tobias; Obel, Carsten; Grønbæk, Kaj

    2015-01-01

    Children with mental disorders like Attention Deficit Hyperactivity Disorder (ADHD) often experience challenges in school as they struggle to maintain their attention. Based on empirical studies conducted in school contexts and together with teachers and ADHD domain professionals, we identified...... design criteria in relation to three core components (sensing, recognizing, and assisting) for designing real time assistive technologies for children with ADHD. Based on these design criteria, we designed the Child Activity Sensing and Training Tool (CASTT), a real time assistive prototype that captures...... activities and assists the child in maintaining attention. From a preliminary evaluation of CASTT with 20 children in several schools, we and found that: 1) it is possible to create a wearable sensor system for children with ADHD that monitors physical and physiological activities in real time; and that 2...

  1. Elementary real and complex analysis

    CERN Document Server

    Shilov, Georgi E

    1996-01-01

    In this book the renowned Russian mathematician Georgi E. Shilov brings his unique perspective to real and complex analysis, an area of perennial interest in mathematics. Although there are many books available on the topic, the present work is specially designed for undergraduates in mathematics, science and engineering. A high level of mathematical sophistication is not required.The book begins with a systematic study of real numbers, understood to be a set of objects satisfying certain definite axioms. The concepts of a mathematical structure and an isomorphism are introduced in Chapter 2,

  2. Real Analysis A Historical Approach

    CERN Document Server

    Stahl, Saul

    2011-01-01

    A provocative look at the tools and history of real analysis This new edition of Real Analysis: A Historical Approach continues to serve as an interesting read for students of analysis. Combining historical coverage with a superb introductory treatment, this book helps readers easily make the transition from concrete to abstract ideas. The book begins with an exciting sampling of classic and famous problems first posed by some of the greatest mathematicians of all time. Archimedes, Fermat, Newton, and Euler are each summoned in turn, illuminating the utility of infinite, power, and trigonome

  3. Comparison of Gull Feces-specific Assays Targeting the 16S rRNA Gene of Catellicoccus Marimammalium and Streptococcus spp.

    Science.gov (United States)

    Two novel gull-specific qPCR assays were developed using 16S rRNA gene sequences from gull fecal clone libraries: a SYBR-green-based assay targeting Streptococcus spp. (i.e., gull3) and a TaqMan qPCR assay targeting Catellicoccus marimammalium (i.e., gull4). The main objectives ...

  4. Real Time Sonic Boom Display

    Science.gov (United States)

    Haering, Ed

    2014-01-01

    This presentation will provide general information about sonic boom mitigation technology to the public in order to supply information to potential partners and licensees. The technology is a combination of flight data, atmospheric data and terrain information implemented into a control room real time display for flight planning. This research is currently being performed and as such, any results and conclusions are ongoing.

  5. History as the Real Thing.

    Science.gov (United States)

    Witten, Sue K.

    1989-01-01

    Describes several ways of using journals, diaries, historical documents, and photographs found in attics or closets to provide stimulating classroom activities. Suggests ways of using the accounts and experiences of real people (preferably the ancestors of students) to enliven history and create interest in historical topics. (GEA)

  6. Computer Screen or Real Life?

    DEFF Research Database (Denmark)

    Suurmets, Seidi; Clement, Jesper

    Aims: In many disciplines, including consumer and marketing research, the findings from eyetracking studies in lab settings are often generalized to natural environments. While studies in the real world have become more common with a statement on increased external validity, the degree to which t...

  7. Lagoa Real design - Mineral engineering

    International Nuclear Information System (INIS)

    Forman, J.M.A.

    1982-01-01

    This paper presents the works realized, in course and to realize of Lagoa Real Design, including the works for implantation of Mineral-Industrial complex with the production capacity of 1.000 ton of U sub(3) O sub(8) per year from 1988. (author)

  8. Uncertainty in the Real World

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 4; Issue 2. Uncertainty in the Real World - Fuzzy Sets. Satish Kumar. General Article Volume 4 Issue 2 February 1999 pp 37-47. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/004/02/0037-0047 ...

  9. Equations for formally real meadows

    NARCIS (Netherlands)

    Bergstra, J.A.; Bethke, I.; Ponse, A.

    2015-01-01

    We consider the signatures Σm = (0,1,−,+,⋅,−1)  of meadows and (Σm,s)  of signed meadows. We give two complete axiomatizations of the equational theories of the real numbers with respect to these signatures. In the first case, we extend the axiomatization of zero-totalized fields by a single axiom

  10. Real time automatic scene classification

    NARCIS (Netherlands)

    Verbrugge, R.; Israël, Menno; Taatgen, N.; van den Broek, Egon; van der Putten, Peter; Schomaker, L.; den Uyl, Marten J.

    2004-01-01

    This work has been done as part of the EU VICAR (IST) project and the EU SCOFI project (IAP). The aim of the first project was to develop a real time video indexing classification annotation and retrieval system. For our systems, we have adapted the approach of Picard and Minka [3], who categorized

  11. Are real teams healthy teams?

    NARCIS (Netherlands)

    Buljac, M.; van Woerkom, M.; van Wijngaarden, P.

    2013-01-01

    This study examines the impact of real-team--as opposed to a team in name only--characteristics (i.e., team boundaries, stability of membership, and task interdependence) on team processes (i.e., team learning and emotional support) and team effectiveness in the long-term care sector. We employed a

  12. Real Time Conference 2016 Overview

    Science.gov (United States)

    Luchetta, Adriano

    2017-06-01

    This is a special issue of the IEEE Transactions on Nuclear Science containing papers from the invited, oral, and poster presentation of the 20th Real Time Conference (RT2016). The conference was held June 6-10, 2016, at Centro Congressi Padova “A. Luciani,” Padova, Italy, and was organized by Consorzio RFX (CNR, ENEA, INFN, Università di Padova, Acciaierie Venete SpA) and the Istituto Nazionale di Fisica Nucleare. The Real Time Conference is multidisciplinary and focuses on the latest developments in real-time techniques in high-energy physics, nuclear physics, astrophysics and astroparticle physics, nuclear fusion, medical physics, space instrumentation, nuclear power instrumentation, general radiation instrumentation, and real-time security and safety. Taking place every second year, it is sponsored by the Computer Application in Nuclear and Plasma Sciences technical committee of the IEEE Nuclear and Plasma Sciences Society. RT2016 attracted more than 240 registrants, with a large proportion of young researchers and engineers. It had an attendance of 67 students from many countries.

  13. Alice in the Real World

    Science.gov (United States)

    Parker, Tom

    2012-01-01

    As a fifth-grade mathematics teacher, the author tries to create authentic problem-solving activities that connect to the world in which his students live. He discovered a natural connection to his students' real world at a computer camp. A friend introduced him to Alice, a computer application developed at Carnegie Mellon, under the leadership of…

  14. Real-time process algebra

    NARCIS (Netherlands)

    Bergstra, J.A.; Baeten, J.C.M.

    1991-01-01

    We describe an axiom system ACPp that incorporates real timed actions. Many examples are provided in order to explain the intuitive contents of the notation. ACP p is a generalisation of ACP. This implies that some of the axioms have to be relaxed and that ACP can be recovered as a special case from

  15. Foley Sounds vs Real Sounds

    DEFF Research Database (Denmark)

    Trento, Stefano; Götzen, Amalia De

    2011-01-01

    This paper is an initial attempt to study the world of sound effects for motion pictures, also known as Foley sounds. Throughout several audio and audio-video tests we have compared both Foley and real sounds originated by an identical action. The main purpose was to evaluate if sound effects are...... applications of sound design such as advertisement or soundtracks for movies....

  16. Uncertainty in the Real World

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 4; Issue 2. Uncertainty in the Real World - Fuzzy Sets. Satish Kumar. General Article Volume 4 Issue 2 February 1999 pp 37-47. Fulltext. Click here to view fulltext PDF. Permanent link: http://www.ias.ac.in/article/fulltext/reso/004/02/0037-0047 ...

  17. Real time freeway incident detection.

    Science.gov (United States)

    2014-04-01

    The US Department of Transportation (US-DOT) estimates that over half of all congestion : events are caused by highway incidents rather than by rush-hour traffic in big cities. Real-time : incident detection on freeways is an important part of any mo...

  18. Real Time Control on Firewire

    NARCIS (Netherlands)

    Zhang, Yuchen

    2004-01-01

    The goal of this project is to get insight into the use of Firewire as a field bus for real-time control. A characterization of Firewire's asynchronous transmission has been made by testing the point-to-point roundtrip in a 3-node Firewire network. The results show Firewire's asynchronous

  19. ISTTOK real-time architecture

    Energy Technology Data Exchange (ETDEWEB)

    Carvalho, Ivo S., E-mail: ivoc@ipfn.ist.utl.pt; Duarte, Paulo; Fernandes, Horácio; Valcárcel, Daniel F.; Carvalho, Pedro J.; Silva, Carlos; Duarte, André S.; Neto, André; Sousa, Jorge; Batista, António J.N.; Hekkert, Tiago; Carvalho, Bernardo B.

    2014-03-15

    Highlights: • All real-time diagnostics and actuators were integrated in the same control platform. • A 100 μs control cycle was achieved under the MARTe framework. • Time-windows based control with several event-driven control strategies implemented. • AC discharges with exception handling on iron core flux saturation. • An HTML discharge configuration was developed for configuring the MARTe system. - Abstract: The ISTTOK tokamak was upgraded with a plasma control system based on the Advanced Telecommunications Computing Architecture (ATCA) standard. This control system was designed to improve the discharge stability and to extend the operational space to the alternate plasma current (AC) discharges as part of the ISTTOK scientific program. In order to accomplish these objectives all ISTTOK diagnostics and actuators relevant for real-time operation were integrated in the control system. The control system was programmed in C++ over the Multi-threaded Application Real-Time executor (MARTe) which provides, among other features, a real-time scheduler, an interrupt handler, an intercommunications interface between code blocks and a clearly bounded interface with the external devices. As a complement to the MARTe framework, the BaseLib2 library provides the foundations for the data, code introspection and also a Hypertext Transfer Protocol (HTTP) server service. Taking advantage of the modular nature of MARTe, the algorithms of each diagnostic data processing, discharge timing, context switch, control and actuators output reference generation, run on well-defined blocks of code named Generic Application Module (GAM). This approach allows reusability of the code, simplified simulation, replacement or editing without changing the remaining GAMs. The ISTTOK control system GAMs run sequentially each 100 μs cycle on an Intel{sup ®} Q8200 4-core processor running at 2.33 GHz located in the ATCA crate. Two boards (inside the ATCA crate) with 32 analog

  20. Making Real-Time Data "Real" for General Interest Users

    Science.gov (United States)

    Hotaling, L.

    2003-04-01

    Helping educators realize the benefits of integrating technology into curricula to effectively engage student learning and improve student achievement, particularly in science and mathematics, is the core mission of the Center for Improved Engineering and Science Education (CIESE). To achieve our mission, we focus on projects utilizing real-time data available from the Internet, and collaborative projects utilizing the Internet's potential to reach peers and experts around the world. As a member of the Mid-Atlantic Center for Ocean Sciences Education Excellence (COSEE), the Center for Improved Engineering and Science Education (CIESE), is committed to delivering relevant ocean science education to diverse audiences, including K-12 teachers, students, coastal managers, families and tourists. The highest priority of the Mid-Atlantic COSEE is to involve scientists and educators in the translation of data and information from the coastal observatories into instructional materials and products usable by educators and the public. A combination of three regional observing systems, the New Jersey Shelf Observing System (NJSOS), Chesapeake Bay Observing System (CBOS), and the York River observing system will provide the scientific backbone for an integrated program of science and education that improves user access to, and understanding of, modern ocean science and how it affects our daily lives. At present, the Mid-Atlantic COSEE offers three projects that enable users to apply and validate scientific concepts to real world situations. (1) The Gulf Stream Voyage is an online multidisciplinary project that utilizes both real-time data and primary source materials to help guide students to discover the science and history of the Gulf Stream current. (2) C.O.O.L. Classroom is an online project that utilizes concepts and real-time data collected through the NJSOS. The C.O.O.L. Classroom is based on the concept of the Rutgers-IMCS Coastal Ocean Observation Laboratory, a real

  1. Development of a multiplex real-time PCR assay for phylogenetic analysis of Uropathogenic Escherichia coli.

    Science.gov (United States)

    Hasanpour, Mojtaba; Najafi, Akram

    2017-06-01

    Uropathogenic Escherichia coli (UPEC) is among major pathogens causing 80-90% of all episodes of urinary tract infections (UTIs). Recently, E. coli strains are divided into eight main phylogenetic groups including A, B1, B2, C, D, E, F, and clade I. This study was aimed to develop a rapid, sensitive, and specific multiplex real time PCR method capable of detecting phylogenetic groups of E. coli strains. This study was carried out on E. coli strains (isolated from the patient with UTI) in which the presence of all seven target genes had been confirmed in our previous phylogenetic study. An EvaGreen-based singleplex and multiplex real-time PCR with melting curve analysis was designed for simultaneous detection and differentiation of these genes. The primers were selected mainly based on the production of amplicons with melting temperatures (T m ) ranging from 82°C to 93°C and temperature difference of more than 1.5°C between each peak.The multiplex real-time PCR assays that have been developed in the present study were successful in detecting the eight main phylogenetic groups. Seven distinct melting peaks were discriminated, with Tm value of 93±0.8 for arpA, 89.2±0.1for chuA, 86.5±0.1 for yjaA, 82.3±0.2 for TspE4C2, 87.8±0.1for trpAgpC, 85.4±0.6 for arpAgpE genes, and 91±0.5 for the internal control. To our knowledge, this study is the first melting curve-based real-time PCR assay developed for simultaneous and discrete detection of these seven target genes. Our findings showed that this assay has the potential to be a rapid, reliable and cost-effective alternative for routine phylotyping of E. coli strains. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Real time psychrometric data collection

    International Nuclear Information System (INIS)

    McDaniel, K.H.

    1996-01-01

    Eight Mine Weather Stations (MWS) installed at the Waste Isolation Pilot Plant (WIPP) to monitor the underground ventilation system are helping to simulate real-time ventilation scenarios. Seasonal weather extremes can result in variations of Natural Ventilation Pressure (NVP) which can significantly effect the ventilation system. The eight MWS(s) (which previously collected and stored temperature, barometric pressure and relative humidity data for subsequent NVP calculations) were upgraded to provide continuous real-time data to the site wide Central monitoring System. This data can now be utilized by the ventilation engineer to create realtime ventilation simulations and trends which assist in the prediction and mitigation of NVP and psychrometric related events

  3. Quantitative (real-time) PCR

    International Nuclear Information System (INIS)

    Denman, S.E.; McSweeney, C.S.

    2005-01-01

    Many nucleic acid-based probe and PCR assays have been developed for the detection tracking of specific microbes within the rumen ecosystem. Conventional PCR assays detect PCR products at the end stage of each PCR reaction, where exponential amplification is no longer being achieved. This approach can result in different end product (amplicon) quantities being generated. In contrast, using quantitative, or real-time PCR, quantification of the amplicon is performed not at the end of the reaction, but rather during exponential amplification, where theoretically each cycle will result in a doubling of product being created. For real-time PCR, the cycle at which fluorescence is deemed to be detectable above the background during the exponential phase is termed the cycle threshold (Ct). The Ct values obtained are then used for quantitation, which will be discussed later

  4. Routines, rigidity and real estate

    OpenAIRE

    Dooley, Kenneth

    2017-01-01

    Finding ways to reduce the environmental impact of the existing building stock is an important element in climate change mitigation. This article examines environmentally focused organisational innovations in the corporate real estate industry. Organisational innovations are often overlooked as they cause considerable disruption to the daily routines of employees. In this article, the focal organisational innovation is the adoption of activity-based working. The study aims to uncover the barr...

  5. Real-time holographic endoscopy

    Science.gov (United States)

    Smigielski, Paul; Albe, Felix; Dischli, Bernard

    1992-08-01

    Some new experiments concerning holographic endoscopy are presented. The quantitative measurements of deformations of objects are obtained by the double-exposure and double- reference beam method, using either a cw-laser or a pulsed laser. Qualitative experiments using an argon laser with time-average holographic endoscopy are also presented. A video film on real-time endoscopic holographic interferometry was recorded with the help of a frequency-doubled YAG-laser working at 25 Hz for the first time.

  6. [Real time 3D echocardiography

    Science.gov (United States)

    Bauer, F.; Shiota, T.; Thomas, J. D.

    2001-01-01

    Three-dimensional representation of the heart is an old concern. Usually, 3D reconstruction of the cardiac mass is made by successive acquisition of 2D sections, the spatial localisation and orientation of which require complex guiding systems. More recently, the concept of volumetric acquisition has been introduced. A matricial emitter-receiver probe complex with parallel data processing provides instantaneous of a pyramidal 64 degrees x 64 degrees volume. The image is restituted in real time and is composed of 3 planes (planes B and C) which can be displaced in all spatial directions at any time during acquisition. The flexibility of this system of acquisition allows volume and mass measurement with greater accuracy and reproducibility, limiting inter-observer variability. Free navigation of the planes of investigation allows reconstruction for qualitative and quantitative analysis of valvular heart disease and other pathologies. Although real time 3D echocardiography is ready for clinical usage, some improvements are still necessary to improve its conviviality. Then real time 3D echocardiography could be the essential tool for understanding, diagnosis and management of patients.

  7. Real-time reverse-transcriptase polymerase chain reaction for the rapid detection of Salmonella using invA primers.

    Science.gov (United States)

    D'Souza, Doris H; Critzer, Faith J; Golden, David A

    2009-11-01

    Recent outbreaks of Salmonella linked to fresh produce emphasize the need for rapid detection methods to help control the spread of disease. Reverse-transcriptase polymerase chain reaction (RT-PCR) can detect the presence of mRNA (shorter half-life than DNA) with greater potential for detecting viable pathogens. The chromosomally located invA gene required for host invasion by Salmonella is widely used for detection of this pathogen by PCR. Detection of Salmonella was undertaken by real-time RT-PCR (rt-RT-PCR) using newly designed invA gene primers to develop a sensitive and specific assay. Salmonella serovars Typhimurium and Enteritidis were grown (7.68 log(10) CFU/mL) in Luria-Bertani broth overnight at 37 degrees C, and RNA was extracted, followed by rt-RT-PCR with and without SYBR green I and agarose gel electrophoresis. All experiments were replicated at least thrice. Detection for both serovars using traditional RT-PCR was lower ( approximately 10(5) CFU/mL) than rt-RT-PCR (10(3) CFU/mL) by gel electrophoresis. Melt curve analysis showed melt temperatures at 87.5 degrees C with Ct values from 12 to 15 for up to 10(3) CFU/mL and improved to 10(2) CFU/mL after further optimization. Further, addition of RNA internal amplification control constructed using in vitro transcription with a T7 RNA polymerase promoter, to the RT-PCR assay also gave detection limits of 10(2) CFU/mL. Cross-reactivity was not observed against a panel of 21 non-Salmonella bacteria. Heat-inactivated (autoclaved) Salmonella showed faint or no detection by rt-RT-PCR or gel electrophoresis. This method has potential to be applied for the detection of Salmonella serovars in fresh produce and the simultaneous detection of foodborne viral (RNA viruses) and bacterial pathogens in a multiplex format.

  8. Sistemas de tiempo real y sistemas distribuidos de tiempo real

    OpenAIRE

    Pesado, Patricia Mabel; Ramón, Hugo Dionisio; Boracchia, Marcos; Pasini, Ariel C.; De Vito, María Cecilia; Osella Massa, Germán Leandro; Estrebou, César Armando

    2006-01-01

    El proyecto de investigación y desarrollo en Sistemas de Software Distribuido, abarca aspectos que van desde los fundamentos del desarrollo (Técnicas de Ingeniería de Requerimientos, Metodologías de Gestión y Desarrollo de Proyectos, Técnicas de Planificación, Métricas, Normas de Calidad, Web-Systems) hasta la concepción de aplicaciones específicas orientadas a los procesos industriales (Sistemas de Planeamiento de Producción, Control Industrial en tiempo real), procesos de E-government (Voto...

  9. Getting Real Results with Ohio State University Extension’s Real Money. Real World. Program

    Directory of Open Access Journals (Sweden)

    Theresa M. Ferrari

    2010-06-01

    Full Text Available Youth development organizations have a unique opportunity to offer programs that help young people develop financial skills they need to become successful adults. This article describes Ohio State University Extension’s Real Money. Real World. (RMRW and the systematic approach used to evaluate its effectiveness. The RMRW curriculum includes an active, hands-on experience that gives young people the opportunity to make lifestyle and budget choices similar to those they will make as adults. The realistic simulation creates a teachable moment. The outcomes of the statewide evaluation clearly indicate the curriculum accomplishes its goals of raising awareness, changing attitudes, and motivating students to plan for behavior changes concerning financial management, education, and career choices. The article concludes with a discussion of the organizational outcomes of conducting the evaluation.

  10. Scalable Real-Time Negotiation Toolkit

    National Research Council Canada - National Science Library

    Lesser, Victor

    2004-01-01

    ... to implement an adaptive distributed sensor network. These activities involved the development of a distributed soft, real-time heuristic resource allocation protocol, the development of a domain-independent soft, real time agent architecture...

  11. Multidimensional real analysis I differentiation

    CERN Document Server

    Duistermaat, J J; van Braam Houckgeest, J P

    2004-01-01

    Part one of the authors' comprehensive and innovative work on multidimensional real analysis. This book is based on extensive teaching experience at Utrecht University and gives a thorough account of differential analysis in multidimensional Euclidean space. It is an ideal preparation for students who wish to go on to more advanced study. The notation is carefully organized and all proofs are clean, complete and rigorous. The authors have taken care to pay proper attention to all aspects of the theory. In many respects this book presents an original treatment of the subject and it contains man

  12. The real Euthyphro Problem, solved

    OpenAIRE

    Denyer, Nicholas Charles

    2017-01-01

    Let us suppose that God commands all and only those actions that are obligatory, and prohibits all and only those actions that are wrong. We then face questions. Does God command the obligatory because it is obligatory? If so, then why pay attention to God’s commands? Why not look rather to the real source of obligatoriness? Or is it instead that the obligatory is obligatory because commanded by God? If so, then would something else have been obligatory if God had commanded otherwise? And why...

  13. A Study towards Real Time Camera Calibration

    OpenAIRE

    Choudhury, Ragini

    2000-01-01

    Preliminary Report Prepared for the Project VISTEO; This report provides a detailed study of the problem of real time camera calibration. This analysis, based on the study of literature in the area, as well as the experiments carried out on real and synthetic data, is motivated by the requirements of the VISTEO project. VISTEO deals with a fusion of real images and synthetic environments, objects etc in TV video sequences. It thus deals with a challenging and fast growing area in virtual real...

  14. Hard Real-Time Networking on FIrewire

    NARCIS (Netherlands)

    Zhang, Yuchen; Orlic, B.; Visser, P.M.; Broenink, Johannes F.; Marquet, P; McGuire, N; Wurmsdobler, P

    2005-01-01

    This paper investigates the possibility of using standard, low-cost, widely used FireWire as a new generation fieldbus medium for real-time distributed control applications. A real-time software subsys- tem, RT-FireWire was designed that can, in combination with Linux-based real-time operating

  15. Texas Real Estate Curriculum Workshop Summary Report.

    Science.gov (United States)

    Lyon, Robert

    The Texas Real Estate Research Center-Texas Education Agency (TRERC-TEA) curriculum workshop was attended by over 40 participants representing 26 Texas community colleges. These participants divided into eight small groups by real estate specialty area and developed curriculum outlines and learning objectives for the following real estate courses:…

  16. A real time monitoring system

    International Nuclear Information System (INIS)

    Fontanini, Horacio; Galdoz, Erwin

    1989-01-01

    A real time monitoring system is described. It was initially developed to be used as a man-machine interface between a basic principles simulator of the Embalse Nuclear Power Plant and the operators. This simulator is under construction at the Bariloche Atomic Center's Process Control Division. Due to great design flexibility, this system can also be used in real plants. The system is designed to be run on a PC XT or AT personal computer with high resolution graphics capabilities. Three interrelated programs compose the system: 1) Graphics Editor, to build static image to be used as a reference frame where to show dynamically updated data. 2) Data acquisition and storage module. It is a memory resident module to acquire and store data in background. Data can be acquired and stored without interference with the operating system, via serial port or through analog-to-digital converter attached to the personal computer. 3) Display module. It shows the acquired data according to commands received from configuration files prepared by the operator. (Author) [es

  17. Real time analysis under EDS

    Science.gov (United States)

    Schneberk, D.

    1985-07-01

    The analysis component of the Enrichment Diagnostic System (EDS) developed for the Atomic Vapor Laser Isotope Separation Program (AVLIS) at Lawrence Livermore National Laboratory (LLNL) is described. Four different types of analysis are performed on data acquired through EDS: (1) absorption spectroscopy on laser-generated spectral lines, (2) mass spectrometer analysis, (3) general purpose waveform analysis, and (4) separation performance calculations. The information produced from this data includes: measures of particle density and velocity, partial pressures of residual gases, and overall measures of isotope enrichment. The analysis component supports a variety of real-time modeling tasks, a means for broadcasting data to other nodes, and a great degree of flexibility for tailoring computations to the exact needs of the process. A particular data base structure and program flow is common to all types of analysis. Key elements of the analysis component are: (1) a fast access data base which can configure all types of analysis, (2) a selected set of analysis routines, (3) a general purpose data manipulation and graphics package for the results of real time analysis.

  18. Lars and the Real Girl

    Directory of Open Access Journals (Sweden)

    Ted Remington

    2011-04-01

    Full Text Available The 2007 film Lars and the Real Girl challenged viewer’s expectations, not simply through its unusual premise (a man having a relationship with a doll, through its consistent refusal to resolve the plot’s various conflicts through verbal or physical violence. Instead, the plot resolves in a way consistent with what peace researcher Johan Galtung has termed positive transcendence, a means of coming to a resolution that meets the needs of all parties. Although the film’s far-fetched plot makes direct practical application of the characters’ actions unlikely, the film performs a more important role in helping to expand moviegoers’ “horizons of expectation.” By prompting its audience to accept an utterly nonviolent, nonconfrontational resolution to the central conflict of the plot, Lars and the Real Girl subverts the notion that satisfying cinematic endings must involve winners and losers. It helps make room for a greater acceptance of nonviolent resolution of problems in popular film.

  19. Real time analysis under EDS

    International Nuclear Information System (INIS)

    Schneberk, D.

    1985-07-01

    This paper describes the analysis component of the Enrichment Diagnostic System (EDS) developed for the Atomic Vapor Laser Isotope Separation Program (AVLIS) at Lawrence Livermore National Laboratory (LLNL). Four different types of analysis are performed on data acquired through EDS: (1) absorption spectroscopy on laser-generated spectral lines, (2) mass spectrometer analysis, (3) general purpose waveform analysis, and (4) separation performance calculations. The information produced from this data includes: measures of particle density and velocity, partial pressures of residual gases, and overall measures of isotope enrichment. The analysis component supports a variety of real-time modeling tasks, a means for broadcasting data to other nodes, and a great degree of flexibility for tailoring computations to the exact needs of the process. A particular data base structure and program flow is common to all types of analysis. Key elements of the analysis component are: (1) a fast access data base which can configure all types of analysis, (2) a selected set of analysis routines, (3) a general purpose data manipulation and graphics package for the results of real time analysis. Each of these components are described with an emphasis upon how each contributes to overall system capability. 3 figs

  20. Real Time Pricing and the Real Live Firm

    Energy Technology Data Exchange (ETDEWEB)

    Moezzi, Mithra; Goldman, Charles; Sezgen, Osman; Bharvirkar, Ranjit; Hopper, Nicole

    2004-05-26

    Energy economists have long argued the benefits of real time pricing (RTP) of electricity. Their basis for modeling customers response to short-term fluctuations in electricity prices are based on theories of rational firm behavior, where management strives to minimize operating costs and optimize profit, and labor, capital and energy are potential substitutes in the firm's production function. How well do private firms and public sector institutions operating conditions, knowledge structures, decision-making practices, and external relationships comport with these assumptions and how might this impact price response? We discuss these issues on the basis of interviews with 29 large (over 2 MW) industrial, commercial, and institutional customers in the Niagara Mohawk Power Corporation service territory that have faced day-ahead electricity market prices since 1998. We look at stories interviewees told about why and how they respond to RTP, why some customers report that they can't, and why even if they can, they don't. Some firms respond as theorized, and we describe their load curtailment strategies. About half of our interviewees reported that they were unable to either shift or forego electricity consumption even when prices are high ($0.50/kWh). Reasons customers gave for why they weren't price-responsive include implicit value placed on reliability, pricing structures, lack of flexibility in adjusting production inputs, just-in-time practices, perceived barriers to onsite generation, and insufficient time. We draw these observations into a framework that could help refine economic theory of dynamic pricing by providing real-world descriptions of how firms behave and why.

  1. Absence of high-risk HPV 16 and 18 in Chinese patients with oral squamous cell carcinoma and oral potentially malignant disorders.

    Science.gov (United States)

    Chen, Xiao-Jie; Sun, Kai; Jiang, Wei-Wen

    2016-05-20

    The critical role of human papillomavirus (HPV) in cancer has been recognized, but the involvement of HPV in oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMD) is still controversial. The aim of this study was to identify and verify the prevalence of high-risk HPV infection (HPV16 and 18) in Chinese patients with OSCC or OPMD using real-time PCR and DNA sequencing. Paired tissue and serum DNA samples were extracted from 40 Chinese patients with OSCC and 59 with OPMD. A SYBR Green-based real-time PCR assay was developed to detect the E6 gene of HPV16 and HPV18. Suspicious positive samples were then sequenced to eliminate false positives. We found that none of the tissue and serum samples of OSCCs and OPMDs were positive for HPV16 E6 or 18 E6, using both real-time PCR and DNA sequencing. Overall, 3 of 198 (1.52 %) and 7 of 198 (3.54 %) samples were false-positive for HPV16 E6 and HPV18 E6, respectively, using real-time PCR. The lack of HPV16 and HPV18 detected in this study indicates that high-risk HPV 16 and 18 infections are uncommon in Chinese patients with OSCC and OPMD. Real-time PCR followed by DNA sequencing for HPV DNA detection is an effective strategy to rule out false positives.

  2. Development of a versatile tool for the simultaneous differential detection of Pseudomonas savastanoi pathovars by End Point and Real-Time PCR.

    Science.gov (United States)

    Tegli, Stefania; Cerboneschi, Matteo; Libelli, Ilaria Marsili; Santilli, Elena

    2010-05-28

    Pseudomonas savastanoi pv. savastanoi is the causal agent of olive knot disease. The strains isolated from oleander and ash belong to the pathovars nerii and fraxini, respectively. When artificially inoculated, pv. savastanoi causes disease also on ash, and pv. nerii attacks also olive and ash. Surprisingly nothing is known yet about their distribution in nature on these hosts and if spontaneous cross-infections occur. On the other hand sanitary certification programs for olive plants, also including P. savastanoi, were launched in many countries. The aim of this work was to develop several PCR-based tools for the rapid, simultaneous, differential and quantitative detection of these P. savastanoi pathovars, in multiplex and in planta. Specific PCR primers and probes for the pathovars savastanoi, nerii and fraxini of P. savastanoi were designed to be used in End Point and Real-Time PCR, both with SYBR Green or TaqMan chemistries. The specificity of all these assays was 100%, as assessed by testing forty-four P. savastanoi strains, belonging to the three pathovars and having different geographical origins. For comparison strains from the pathovars phaseolicola and glycinea of P. savastanoi and bacterial epiphytes from P. savastanoi host plants were also assayed, and all of them tested always negative. The analytical detection limits were about 5 - 0.5 pg of pure genomic DNA and about 102 genome equivalents per reaction. Similar analytical thresholds were achieved in Multiplex Real-Time PCR experiments, even on artificially inoculated olive plants. Here for the first time a complex of PCR-based assays were developed for the simultaneous discrimination and detection of P. savastanoi pv. savastanoi, pv. nerii and pv. fraxini. These tests were shown to be highly reliable, pathovar-specific, sensitive, rapid and able to quantify these pathogens, both in multiplex reactions and in vivo. Compared with the other methods already available for P. savastanoi, the identification

  3. Introduction to Real Analysis An Educational Approach

    CERN Document Server

    Bauldry, William C

    2011-01-01

    An accessible introduction to real analysis and its connection to elementary calculus Bridging the gap between the development and history of real analysis, Introduction to Real Analysis: An Educational Approach presents a comprehensive introduction to real analysis while also offering a survey of the field. With its balance of historical background, key calculus methods, and hands-on applications, this book provides readers with a solid foundation and fundamental understanding of real analysis. The book begins with an outline of basic calculus, including a close examination of problems illust

  4. Towards phronetic knowledge for strategic planning in corporate real estate management: A real options approach

    NARCIS (Netherlands)

    van Reedt Dortland, Maartje; Voordijk, Johannes T.; Dewulf, Geert P.M.R.

    2014-01-01

    Purpose – The objective of this paper is to provide insights about the potential of real option thinking for corporate real estate management (CREM) from the owner-user perspective. A promising approach to classifying and evaluating flexibility in real estate is the real options approach. Most

  5. Real oscillations of virtual neutrinos

    International Nuclear Information System (INIS)

    Grimus, W.; Stockinger, P.

    1996-01-01

    We study the conditions for neutrino oscillations in a field-theoretical approach by taking into account that only the neutrino production and detection processes, which are localized in space around the coordinates x searrow P and x searrow D , respectively, can be manipulated. In this sense the neutrinos whose oscillations are investigated appear as virtual lines connecting production with detection in the total Feynman graph and all neutrino fields or states to be found in the discussion are mass eigenfields or eigenstates. We perform a thorough examination of the integral over the spatial components of the inner neutrino momentum and show that in the asymptotic limit L=|x searrow D -x searrow P |→∞ the virtual neutrinos become open-quote open-quote real close-quote close-quote and under certain conditions the usual picture of neutrino oscillations emerges without ambiguities. copyright 1996 The American Physical Society

  6. Shopping in the Real World

    DEFF Research Database (Denmark)

    Black, Darren; Clemmensen, Nils Jakob; Skov, Mikael B.

    2009-01-01

    Shopping in the real world is becoming an increasingly interactive experience as stores integrate various technologies to support shoppers. Based on an empirical study of supermarket shoppers, we designed a mobile context-aware system called the Context- Aware Shopping Trolley (CAST). The aim...... of the system is to support shopping in supermarkets through context-awareness and acquiring user attention. Thus, the interactive trolley guides and directs shoppers in the handling and finding of groceries. An empirical evaluation showed that shoppers using CAST adapted in different shopping behavior than...... traditional trolley shoppers by exhibiting a more uniform behavior in terms of product sequence collection and ease of finding products and thus, CAST supported the shopping experience....

  7. SOFC Operation with Real Biogas

    DEFF Research Database (Denmark)

    Hagen, Anke; Winiwarter, Anna; Langnickel, Hendrik

    2017-01-01

    , state‐of‐the‐art SOFCs were studied regarding performance and durability in relation to biogas as fuel and considering important contaminants, specifically sulfur. First, the catalytic behavior in relevant synthetic biogas mixtures was studied and the potential of dry reforming was demonstrated....... Successful long term operation of an SOFC under both, conditions of steam and dry reforming, i.e., addition of steam or CO2 to avoid carbon formation was shown. For the steam reforming case a remarkable period of 3,500 h, hereof 3,000 h in the presence of H2S was achieved. Finally, a real biogas from...... a landfill gas unit was used as fuel. The concept of dry reforming was realized. The SOFC was successfully operated with and in one case even without a specific gas cleaning unit....

  8. The effects of real exchange rate misalignment and real exchange volatility on exports

    OpenAIRE

    Diallo, Ibrahima Amadou

    2011-01-01

    This paper uses panel data cointegration techniques to study the impacts of real exchange rate misalignment and real exchange rate volatility on total exports for a panel of 42 developing countries from 1975 to 2004. The results show that both real exchange rate misalignment and real exchange rate volatility affect negatively exports. The results also illustrate that real exchange rate volatility is more harmful to exports than misalignment. These outcomes are corroborated by estimations on s...

  9. Dalla crisi finanziaria alla crisi reale. ( Financial and real crisis

    Directory of Open Access Journals (Sweden)

    Carlo D'Adda

    2010-03-01

    Full Text Available  The financial crisis of 2008 is put in relation with two bubbles, financial and housing. Easy credit is at the origin of both bubbles. Risky bonds were issued to mobilize mortgages originated in the housing market, and bad bonds entered a fantastic number of institutional and private portfolios all over the world without any perception of the their risk. Unfortunately, bank credit to speculators was abundant and fed the final growth of the bubbles. When fears started to spread, the fall was immediate and expectations impaired very rapidly. The main aggregate demand components were severely curtailed and production fell as well. This is the crisis of the real economy. Fortunately, as opposed to what happened in 1929, the interventions of central banks and governments have been of an unprecedented size. The bail out of insolvent banks in particular has been providential. The exit from the current crisis, however, will surely be slow.  JEL: E44, G01, G10

  10. Real-time scene generator

    Science.gov (United States)

    Lord, Eric; Shand, David J.; Cantle, Allan J.

    1996-05-01

    This paper describes the techniques which have been developed for an infra-red (IR) target, countermeasure and background image generation system working in real time for HWIL and Trial Proving applications. Operation is in the 3 to 5 and 8 to 14 micron bands. The system may be used to drive a scene projector (otherwise known as a thermal picture synthesizer) or for direct injection into equipment under test. The provision of realistic IR target and countermeasure trajectories and signatures, within representative backgrounds, enables the full performance envelope of a missile system to be evaluated. It also enables an operational weapon system to be proven in a trials environment without compromising safety. The most significant technique developed has been that of line by line synthesis. This minimizes the processing delays to the equivalent of 1.5 frames from input of target and sightline positions to the completion of an output image scan. Using this technique a scene generator has been produced for full closed loop HWIL performance analysis for the development of an air to air missile system. Performance of the synthesis system is as follows: 256 * 256 pixels per frame; 350 target polygons per frame; 100 Hz frame rate; and Gouraud shading, simple reflections, variable geometry targets and atmospheric scaling. A system using a similar technique has also bee used for direct insertion into the video path of a ground to air weapon system in live firing trials. This has provided realistic targets without degrading the closed loop performance. Delay of the modified video signal has been kept to less than 5 lines. The technique has been developed using a combination of 4 high speed Intel i860 RISC processors in parallel with the 4000 series XILINX field programmable gate arrays (FPGA). Start and end conditions for each line of target pixels are prepared and ordered in the I860. The merging with background pixels and output shading and scaling is then carried out in

  11. Real-time Pricing in Power Markets

    DEFF Research Database (Denmark)

    Boom, Anette; Schwenen, Sebastian

    We examine welfare e ects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction ....... In the Bertrand case, welfare is the same with all or no consumers on smart meters.......We examine welfare e ects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction...... to satisfy demand from retailers acting on behalf of subscribed customers and from consumers with real-time meters. Increasing the number of consumers on real-time pricing does not always increase welfare since risk-averse consumers dislike uncertain and high prices arising through market power...

  12. Real-time Pricing in Power Markets

    DEFF Research Database (Denmark)

    Boom, Anette; Schwenen, Sebastian

    We examine welfare eects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction t....... In the Bertrand case, welfare is the same with all or no consumers on smart meters.......We examine welfare eects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction...... to satisfy demand from retailers acting on behalf of subscribed customers and from consumers with real-time meters. Increasing the number of consumers on real-time pricing does not always increase welfare since risk-averse consumers dislike uncertain and high prices arising through market power...

  13. O direito real de Kant Kant's real right

    Directory of Open Access Journals (Sweden)

    Aylton Barbieri Durão

    2010-01-01

    Full Text Available No direito real, Kant investiga a propriedade privada de uma substância (o solo e os objetos localizados nele. No estado de natureza, somente ocorre a posse física ou empírica de um objeto externo, a partir do postulado jurídico da razão prática como uma lei permissiva, pois, do contrário, as coisas utilizáveis seriam em-si ou res nullius, mas a posse jurídica ou inteligível depende da posse comum originária do solo (que não se confunde com o comunismo primitivo, para evitar que a propriedade seja uma relação entre pessoas e coisas, como acontece na teoria do trabalho, porque todo direito exige um dever correspondente. Todavia, coisas não podem ter deveres com relação a pessoas, condição seguida da vontade unilateral do primeiro ocupante de querer o objeto e do advento de uma vontade unificada a priori do provo, que só se torna efetiva no estado civil, a única capaz de gerar uma obrigação recíproca entre todos; por isso, a posse inteligível somente é possível no estado civil, embora a posse empírica do estado de natureza tenha a presunção de tornar-se jurídica, quando ocorrer a entrada no estado jurídico e vale por comparação na espera e preparação daquele estado.In real right, Kant investigates the private property of a substance (the soil and the objects on it. In the state of nature the physical or empirical possession of an external object only occurs based on the juridical postulate of the practical reason as a permissive law; otherwise, usable things would be in themselves, or res nullius, but the juridical or intelligible possession depends on the common possession which originates from the soil (and cannot be confounded with primitive communism, to avoid the property to be a relation among people and things, such as what happens in the work theory, because all rights demand a correlated duty, but things cannot have duties towards people, followed by the first occupant's unilateral will to want the

  14. Determinants of Real Estate Licensee Income

    OpenAIRE

    G. Stacy Sirmans; Philip G. Swicegood

    1997-01-01

    This paper examines the factors that influence the income of real estate licensees. An empirical human capital earnings model is developed from a 1995 survey of Florida real estate brokers and salespeople. In seeking to explain earnings of real estate licensees, this study expands from previous studies by measuring several additional human capital components. A number of factors are seen to positively affect licensee income. These include (a) number of hours worked, (b) experience, (c) franch...

  15. A Real time network at home

    OpenAIRE

    Hanssen, F.T.Y.; Jansen, P.G.; Hartel, Pieter H.; Scholten, Johan; Vervoort, Wiek; Karelse, F.

    2001-01-01

    This paper proposes a home network which integrates both real-time and non-real-time capabilities for one coherent, distributed architecture. Such a network is not yet available. Our network will support inexpensive, small appliances as well as more expensive, large appliances. The network is based on a new type of real-time token protocol that uses scheduling to achieve optimal token-routing through the network. Depending on the scheduling algorithm, bandwidth utilisations of 100 percent are...

  16. Equivalences of real submanifolds in complex space.

    OpenAIRE

    ZAITSEV, DMITRI

    2001-01-01

    PUBLISHED We show that for any real-analytic submanifold M in CN there is a proper real-analytic subvariety V contained in M such that for any p ? M \\ V , any real-analytic submanifold M? in CN, and any p? ? M?, the germs of the submanifolds M and M? at p and p? respectively are formally equivalent if and only if they are biholomorphically equivalent. More general results for k-equivalences are also stated and proved.

  17. Technical life cycle of real estates

    OpenAIRE

    Taina Koskelo

    2001-01-01

    Real estate business is developing fast. Development needs are challenging because of interdiciplinariness and diverseness of the branch. The main mission of real estate business is to provide premises for its customers. Designers are planning the buildings, contractors are building them and owners are supplying premises for users. During the use the real estate market needs different kinds of facilitating services directed to users or owner of buildings. Facilities Management services (FM) c...

  18. Controls Over Army Real Property Financial Reporting

    Science.gov (United States)

    2008-03-28

    Controls Over Army Real Property Financial Reporting Report No. D-2008-072 March 28, 2008 Report Documentation Page Form...COVERED 00-00-2008 to 00-00-2008 4. TITLE AND SUBTITLE Controls Over Army Real Property Financial Reporting 5a. CONTRACT NUMBER 5b. GRANT NUMBER...DIRECTOR, DEFENSE FINANCE AND ACCOUNTING SERVICE SUBJECT: Report on Controls Over Army Real Property Financial Reporting ( Report No. D-2008-072

  19. Listing Specialization and Residential Real Estate Licensee

    OpenAIRE

    Leonard V. Zumpano; Ken Johnson; Randy I. Anderson

    2007-01-01

    Earlier research has found that specialization by real estate agents creates economies of scope for real estate firms. So far, however, no research has addressed this issue at the agent level. The question this research seeks to answer is whether specialization in one side of the real estate transaction increases agent income. The most important finding is that specialization has an asymmetric impact on earnings. Specializing in listings positively enhances agent income. In contrast, speciali...

  20. Real Orthogonal Projections as Quantum Logic

    Science.gov (United States)

    Matvejchuk, Marjan

    2017-12-01

    In this paper, we study linear operators on real and complex Euclidean spaces which are real-orthogonal projections. It is a generalization of such standard (complex) orthogonal projections for which only the real part of scalar product vanishes. We can compare some partial order properties of the orthogonal and of the R-orthogonal projections. We prove that the set of all R-orthogonal projections in finite-dimensional complex space is a quantum logic.

  1. Real Options and Human Capital Investment

    OpenAIRE

    Bas Jacobs

    2007-01-01

    This paper extends the standard human capital model with real options. Real options influence investment behavior when risky investments in human capital are irreversible and individuals can affect the timing of the investment. Option values make individuals more reluctant to invest in human capital and, as a result, required returns on the investment increase. Real options may help to explain a larger human capital premium for higher education, smaller responsiveness of higher education inve...

  2. Mobile real time radiography system

    International Nuclear Information System (INIS)

    Vigil, J.; Taggart, D.; Betts, S.

    1997-01-01

    A 450-keV Mobile Real Time Radiography (RTR) System was delivered to Los Alamos National Laboratory (LANL) in January 1996. It was purchased to inspect containers of radioactive waste produced at (LANL). Since its delivery it has been used to radiograph more than 600 drums of radioactive waste at various LANL sites. It has the capability of inspecting waste containers of various sizes from <1-gal. buckets up to standard waste boxes (SWB, dimensions 54.5 in. x 71 in. x 37 in.). It has three independent x-ray acquisition formats. The primary system used is a 12- in. image intensifier, the second is a 36-in. linear diode array (LDA) and the last is an open system. It is fully self contained with on board generator, HVAC, and a fire suppression system. It is on a 53-ft long x 8-ft. wide x 14-ft. high trailer that can be moved over any highway requiring only an easily obtainable overweight permit because it weights ∼38 tons. It was built to conform to industry standards for a cabinet system which does not require an exclusion zone. The fact that this unit is mobile has allowed us to operate where the waste is stored, rather than having to move the waste to a fixed facility

  3. Real time 3D photometry

    Science.gov (United States)

    Fernandez-Balbuena, A. A.; Vazquez-Molini, D.; García-Botella, A.; Romo, J.; Serrano, Ana

    2017-09-01

    The photometry and radiometry measurement is a well-developed field. The necessity of measuring optical systems performance involves the use of several techniques like Gonio-photometry. The Gonio photometers are a precise measurement tool that is used in the lighting area like office, luminaire head car lighting, concentrator /collimator measurement and all the designed and fabricated optical systems that works with light. There is one disadvantage in this kind of measurements that obtain the intensity polar curves and the total flux of the optical system. In the industry, there are good Gonio photometers that are precise and reliable but they are very expensive and the measurement time is long. In industry the cost can be of minor importance but measuring time that is around 30 minutes is of major importance due to trained staff cost. We have designed a system to measure photometry in real time; it consists in a curved screen to get a huge measurement angle and a CCD. The system to be measured projects light onto the screen and the CCD records a video of the screen obtaining an image of the projected profile. A complex calibration permits to trace screen data (x,y,z) to intensity polar curve (I,αγ). This intensity is obtained in candels (cd) with an image + processing time below one second.

  4. DNA internal standard for the quantitative determination of hallucinogenic plants in plant mixtures.

    Science.gov (United States)

    Luciano, Pino; Bertea, Cinzia M; Temporale, Giovanni; Maffei, Massimo E

    2007-12-01

    Here, we show a new, simple, and rapid SYBR Green-based Real-Time PCR assay for the quantification of hallucinogenic plants in plant mixtures. As a test plant, Salvia divinorum Epling & Játiva-M., a perennial herb belonging to the Lamiaceae family able to induce hallucinations, changes in perception, or other psychologically induced changes with similar potency as LSD, was used. The method was tested on seven mixtures 100/0%, 80/20%, 60/40%, 40/60%, 20/80%, 10/90%, 0/100% (w/w) S. divinorum versus a non-hallucinogenic plant, Salvia officinalis. Total DNA was extracted from samples and quantified by Real-Time PCR. Arabidopsis thaliana genomic DNA was added, as internal standard, at the beginning of each extraction. A new formula for the interpretation of Real-Time PCR data, based on the relative quantification of DNA extracted from mixture versus a reference DNA extracted from a known amount of pure S. divinorum, was developed. The results of this work show an almost perfect correspondence between Real-Time PCR-calculated weight and the weight estimated by an analytical weighted method, proving the effectiveness of this method for the quantitative analysis of a given species in a plant mixture.

  5. Accounting costs of transactions in real estate

    DEFF Research Database (Denmark)

    Stubkjær, Erik

    2005-01-01

    The costs of transactions in real estate is of importance for households, for investors, for statistical services, for governmental and international bodies concerned with the efficient delivery of basic state functions, as well as for research. The paper takes a multi-disciplinary approach...... in relating theoretical conceptualizations of transaction costs to national accounting and further to the identification and quantification of actions on units of real estate. The notion of satellite accounting of the System of National Accounts is applied to the segment of society concerned with changes...... in real estate. The paper ends up with an estimate of the cost of a major real property transaction in Denmark....

  6. Real-time Pricing in Power Markets

    DEFF Research Database (Denmark)

    Boom, Anette; Schwenen, Sebastian

    We examine welfare eects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction...... to satisfy demand from retailers acting on behalf of subscribed customers and from consumers with real-time meters. Increasing the number of consumers on real-time pricing does not always increase welfare since risk-averse consumers dislike uncertain and high prices arising through market power...

  7. Real-time Pricing in Power Markets

    DEFF Research Database (Denmark)

    Boom, Anette; Schwenen, Sebastian

    We examine welfare e ects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction...... to satisfy demand from retailers acting on behalf of subscribed customers and from consumers with real-time meters. Increasing the number of consumers on real-time pricing does not always increase welfare since risk-averse consumers dislike uncertain and high prices arising through market power...

  8. Controls Over Army Real Property Financial Reporting

    National Research Council Canada - National Science Library

    Marsh, Patricia A

    2008-01-01

    .... A well-designed process for reporting real property on financial statements employs standard data elements and transaction processes, consistent internal controls, and efficient transaction entry...

  9. Real-time Pricing in Power Markets

    DEFF Research Database (Denmark)

    Boom, Anette; Schwenen, Sebastian

    to satisfy demand from retailers acting on behalf of subscribed customers and from consumers with real-time meters. Increasing the number of consumers on real-time pricing does not always increase welfare since risk-averse consumers dislike uncertain and high prices arising through market power......We examine welfare eects of real-time pricing in electricity markets. Before stochastic energy demand is known, competitive retailers contract with nal consumers who exogenously do not have real-time meters. After demand is realized, two electricity generators compete in a uniform price auction...

  10. Research in Distributed Real-Time Systems

    Science.gov (United States)

    Mukkamala, R.

    1997-01-01

    This document summarizes the progress we have made on our study of issues concerning the schedulability of real-time systems. Our study has produced several results in the scalability issues of distributed real-time systems. In particular, we have used our techniques to resolve schedulability issues in distributed systems with end-to-end requirements. During the next year (1997-98), we propose to extend the current work to address the modeling and workload characterization issues in distributed real-time systems. In particular, we propose to investigate the effect of different workload models and component models on the design and the subsequent performance of distributed real-time systems.

  11. Real issue with nuclear power

    International Nuclear Information System (INIS)

    Simpson, J.W.

    1976-01-01

    The voter referendums on nuclear power planned in some states can affect the energy supply and economic health of the public at large more than it affects the industry that provides nuclear power, the author states. He makes the point that those responsible for energy supplies in the U. S.--the President and all relevant Federal agencies, the majority of Congress, the national utility industry, major laboratories, universities and consulting firms, and other energy industries--all favor nuclear power. The complex U.S. energy situation is reviewed, and the hope of alternative energy sources, practice of energy conservation, and benefits of nuclear power are summarized. Specifically, the California Initiative and its three conditions which it says should dictate the future of nuclear power are reviewed. The author does not believe that the reasons that are usually given in opposing nuclear power are the real reasons. He states that ''it seems clear that the principal philosophy behind the initiatives is one of halting economic growth by striking at the energy source that would make that growth possible.'' Attention is called to the morality of nuclear power by asking where is the morality: in leaving future generations an insufficient amount of energy, limiting their abilities to solve the economic and employment problems; in squandering our finite supply of fossil fuels while ignoring nuclear fuels; in forcing the nation into further dependence on unpredictable foreign nations for its energy supply; in expecting other states to provide California with the energy that it does not want to generate itself; and in allowing an arbitrary limit on growth to be set by groups of political activists

  12. 12 CFR 541.21 - Nonresidential real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Nonresidential real estate. 541.21 Section 541... REGULATIONS AFFECTING FEDERAL SAVINGS ASSOCIATIONS § 541.21 Nonresidential real estate. The terms nonresidential real estate or nonresidential real property mean real estate that is not residential real estate...

  13. Debt capacity of real estate collateral

    NARCIS (Netherlands)

    Giambona, E.; Golec, J.; Schwienbacher, A.

    2014-01-01

    We study whether real estate assets have a greater positive influence on firm leverage than other tangible assets. Using a large sample of COMPUSTAT firms, we find a significant positive relation between tangibility and leverage in general, and the relation is strongest for real estate collateral.

  14. Obtaining Your License: Careers in Real Estate.

    Science.gov (United States)

    Lyon, Robert

    Two steps are required to obtain a real estate salesperson's license in Texas: (1) selecting a broker to serve as an advisor, and (2) meeting personal requirements (at least 18 years old, a Texas resident, completion of a minimum of 12 semester hours of real estate and related courses, application, acceptable score on state exam, and payment of…

  15. Corporate real estate and competitive strategy

    NARCIS (Netherlands)

    Bossink, B.A.G.; Singer, B.P.; Vande Putte, H.J.M.

    2007-01-01

    Purpose The purpose of this paper is to investigate how organisations use a corporate real estate strategy to support their competitive strategy. It provides a theoretical and empirical overview and analysis of effective combinations of firms' real estate and competitive strategies.

  16. Real hypersurfaces of a complex space form

    Indian Academy of Sciences (India)

    In this paper we are interested in obtaining a condition under which a compact real hypersurface of a complex projective space C P n is a geodesic sphere. We also study the question as to whether the characteristic vector field of a real hypersurface of the complex projective space C P n is harmonic, and show that the ...

  17. Real hypersurfaces of a complex space form

    Indian Academy of Sciences (India)

    Abstract. In this paper we are interested in obtaining a condition under which a com- pact real hypersurface of a complex projective space CPn is a geodesic sphere. We also study the question as to whether the characteristic vector field of a real hypersurface of the complex projective space CPn is harmonic, and show that ...

  18. Real-time communication protocols: an overview

    NARCIS (Netherlands)

    Hanssen, F.T.Y.; Jansen, P.G.

    2003-01-01

    This paper describes several existing data link layer protocols that provide real-time capabilities on wired networks, focusing on token-ring and Carrier Sense Multiple Access based networks. Existing modifications to provide better real-time capabilities and performance are also described. Finally

  19. A Real time network at home

    NARCIS (Netherlands)

    Hanssen, F.T.Y.; Jansen, P.G.; Hartel, Pieter H.; Scholten, Johan; Vervoort, Wiek; Karelse, F.

    2001-01-01

    This paper proposes a home network which integrates both real-time and non-real-time capabilities for one coherent, distributed architecture. Such a network is not yet available. Our network will support inexpensive, small appliances as well as more expensive, large appliances. The network is based

  20. Real time programming environment for Windows

    Energy Technology Data Exchange (ETDEWEB)

    LaBelle, D.R. [LaBelle (Dennis R.), Clifton Park, NY (United States)

    1998-04-01

    This document provides a description of the Real Time Programming Environment (RTProE). RTProE tools allow a programmer to create soft real time projects under general, multi-purpose operating systems. The basic features necessary for real time applications are provided by RTProE, leaving the programmer free to concentrate efforts on his specific project. The current version supports Microsoft Windows{trademark} 95 and NT. The tasks of real time synchronization and communication with other programs are handled by RTProE. RTProE includes a generic method for connecting a graphical user interface (GUI) to allow real time control and interaction with the programmer`s product. Topics covered in this paper include real time performance issues, portability, details of shared memory management, code scheduling, application control, Operating System specific concerns and the use of Computer Aided Software Engineering (CASE) tools. The development of RTProE is an important step in the expansion of the real time programming community. The financial costs associated with using the system are minimal. All source code for RTProE has been made publicly available. Any person with access to a personal computer, Windows 95 or NT, and C or FORTRAN compilers can quickly enter the world of real time modeling and simulation.

  1. Kolmogorov Complexity Theory over the reals

    NARCIS (Netherlands)

    Ziegler, M.; Koolen, W.M.

    2008-01-01

    Kolmogorov Complexity constitutes an integral part of computability theory, information theory, and computational complexity theory—in the discrete setting of bits and Turing machines. Over real numbers, on the other hand, the BSS-machine (aka real-RAM) has been established as a major model of

  2. Storm real-time processing cookbook

    CERN Document Server

    Anderson, Quinton

    2013-01-01

    A Cookbook with plenty of practical recipes for different uses of Storm.If you are a Java developer with basic knowledge of real-time processing and would like to learn Storm to process unbounded streams of data in real time, then this book is for you.

  3. Quantitative real-time imaging of glutathione

    Science.gov (United States)

    Glutathione plays many important roles in biological processes; however, the dynamic changes of glutathione concentrations in living cells remain largely unknown. Here, we report a reversible reaction-based fluorescent probe—designated as RealThiol (RT)—that can quantitatively monitor the real-time ...

  4. Transitions toward a Semantics for Real Possibility

    NARCIS (Netherlands)

    Rumberg, A.S.

    2016-01-01

    The aim of this thesis consists in developing a semantics for the notion of real possibility. Real possibilities are alternative possibilities for the future in an indeterministic world. In our investigations, we depart from the theory of branching time. The task of providing a semantics for the

  5. Real time refractive index measurement by ESPI

    International Nuclear Information System (INIS)

    Torroba, R.; Joenathan, C.

    1991-01-01

    In this paper a method to measure refractive index variations in real time is reported. A technique to introduce reference fringes in real time is discussed. Both the theoretical and experimental results are presented and an example with phase shifting is given. (author). 8 refs, 5 figs

  6. 36 CFR 1207.31 - Real property.

    Science.gov (United States)

    2010-07-01

    ... original purchase to the fair market value of the property. However, in those situations where a grantee or... purchase of the real property to the current fair market value of the property. ... GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards § 1207.31 Real property. (a) Title...

  7. 44 CFR 13.31 - Real property.

    Science.gov (United States)

    2010-10-01

    ... original purchase to the fair market value of the property. However, in those situations where a grantee or... purchase of the real property to the current fair market value of the property. ... LOCAL GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards § 13.31 Real property. (a...

  8. An Analysis of the Real Estate Occupation.

    Science.gov (United States)

    Harestad, Clifford E.; And Others

    The general purpose of the occupational analysis is to provide workable, basic information dealing with the many and varied duties performed in the real estate occupation. It represents an accounting of what must be done, but not how the entire cycle of a residential sale is completed through the services of a real estate person and the…

  9. Matlab Code for Sorted Real Schur Forms

    NARCIS (Netherlands)

    Brandts, J.H.

    2001-01-01

    In Matlab, there exists a standard command to generate a real Schur form, and another command transforms a real Schur form into a complex one. In Golub and Van Loan (1996), a Matlab-like routine is sketched that sorts a complex Schur form: given a target value ? in the complex plane, the diagonal

  10. Learning from Dealing with Real World Problems

    Science.gov (United States)

    Akcay, Hakan

    2017-01-01

    The purpose of this article is to provide an example of using real world issues as tools for science teaching and learning. Using real world issues provides students with experiences in learning in problem-based environments and encourages them to apply their content knowledge to solving current and local problems.

  11. Commercial real estate analysis and investments

    NARCIS (Netherlands)

    Geltner, D.; Miller, N.; Clayton, J.; Eichholtz, P.M.A.

    2013-01-01

    The well-known and respected authorship team of Geltner and Miller bring you a new edition of what has become the undisputed and authoritative resource on commercial real estate investment. Streamlined and completely updated with expanded coverage of corporate and international real estate

  12. Validation of RNAi by real time PCR

    DEFF Research Database (Denmark)

    Josefsen, Knud; Lee, Ying Chiu

    2011-01-01

    Real time PCR is the analytic tool of choice for quantification of gene expression, while RNAi is concerned with downregulation of gene expression. Together, they constitute a powerful approach in any loss of function studies of selective genes. We illustrate here the use of real time PCR to verify...

  13. Moments of real relationship in psychoanalytic supervision.

    Science.gov (United States)

    Watkins, C Edward

    2012-09-01

    What role does the real relationship play in psychoanalytic supervision? While the real relationship's role has long been and continues to be considered with regard to psychoanalysis, it has received virtually no attention in the supervision literature. In this paper, using Horney's construct of the real self as conceptual anchor, I attempt to: (1) situate the real relationship squarely within the borders of the psychoanalytic supervision relationship; (2) examine the relevance of real relationship phenomena for the supervision experience; (3) provide some simple, ordinary yet meaningful examples of case dialogue that illustrate moments of real relationship in supervision; and (4) introduce the concept of real relationship rupture and consider its potential ramifications for and impact upon the supervisor-supervisee relationship. Just as ruptures can occur in the supervisory alliance, I propose that ruptures can also transpire in the supervisory real relationship, have the potential to reverberate throughout the entirety of the supervision experience, and depending upon how they are handled, can prove either constructive and relationally energizing and enlivening or enervating and eviscerating to supervision process and outcome.

  14. Real balance effects, timing, and equilibrium determination

    NARCIS (Netherlands)

    Stoltenberg, C.A.

    2012-01-01

    By assuming that money balances at the beginning instead of at the end of the period provide transaction services, standard results on nominal and real determinacy in monetary models are overturned. The key is that predetermined real money balances can be a state variable. Whereas the determination

  15. CPR: A Real Lifesaver (For Kids)

    Science.gov (United States)

    ... First Aid & Safety Doctors & Hospitals Videos Recipes for Kids Kids site Sitio para niños How the Body Works ... English Español CPR: A Real Lifesaver KidsHealth / For Kids / CPR: A Real Lifesaver What's in this article? ...

  16. Multiprocessor scheduling for real-time systems

    CERN Document Server

    Baruah, Sanjoy; Buttazzo, Giorgio

    2015-01-01

    This book provides a comprehensive overview of both theoretical and pragmatic aspects of resource-allocation and scheduling in multiprocessor and multicore hard-real-time systems.  The authors derive new, abstract models of real-time tasks that capture accurately the salient features of real application systems that are to be implemented on multiprocessor platforms, and identify rules for mapping application systems onto the most appropriate models.  New run-time multiprocessor scheduling algorithms are presented, which are demonstrably better than those currently used, both in terms of run-time efficiency and tractability of off-line analysis.  Readers will benefit from a new design and analysis framework for multiprocessor real-time systems, which will translate into a significantly enhanced ability to provide formally verified, safety-critical real-time systems at a significantly lower cost.

  17. LabVIEW Real-Time

    CERN Multimedia

    CERN. Geneva; Flockhart, Ronald Bruce; Seppey, P

    2003-01-01

    With LabVIEW Real-Time, you can choose from a variety of RT Series hardware. Add a real-time data acquisition component into a larger measurement and automation system or create a single stand-alone real-time solution with data acquisition, signal conditioning, motion control, RS-232, GPIB instrumentation, and Ethernet connectivity. With the various hardware options, you can create a system to meet your precise needs today, while the modularity of the system means you can add to the solution as your system requirements grow. If you are interested in Reliable and Deterministic systems for Measurement and Automation, you will profit from this seminar. Agenda: Real-Time Overview LabVIEW RT Hardware Platforms - Linux on PXI Programming with LabVIEW RT Real-Time Operating Systems concepts Timing Applications Data Transfer

  18. GetReal : from efficacy in clinical trials to relative effectiveness in the real world

    NARCIS (Netherlands)

    Egger, Matthias; Moons, Karel G M; Fletcher, Christine

    The GetReal consortium ("incorporating real-life data into drug development") addresses the efficacy-effectiveness gap that opens between the data from well-controlled randomized trials in selected patient groups submitted to regulators and the real-world evidence on effectiveness and safety of

  19. 75 FR 74620 - Real Estate Settlement Procedures Act (RESPA): Home Warranty Companies' Payments to Real Estate...

    Science.gov (United States)

    2010-12-01

    ... certain provisions of RESPA as applied to the payment of fees to real estate brokers and agents by home... not compensable under RESPA, a real estate broker or agent (or other person in a position to refer... primary services provided by the real estate broker or agent, if the services are not nominal, and the...

  20. 75 FR 36271 - Real Estate Settlement Procedures Act (RESPA): Home Warranty Companies' Payments to Real Estate...

    Science.gov (United States)

    2010-06-25

    ... RESPA and HUD's implementing RESPA regulations, services performed by real estate brokers and agents as..., necessary and distinct from the primary services provided by the real estate broker or agent, the services... CFR 3500.2. Accordingly, the framework for compensation of real estate brokers and agents for services...

  1. Brilliant green-based PVC membrane electrode for the ...

    African Journals Online (AJOL)

    Bulletin of the Chemical Society of Ethiopia. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 2, No 1 (1988) >. Log in or Register to get access to full text downloads.

  2. The ALMA Real Time Control System

    Science.gov (United States)

    Kern, Jeffrey S.; Juerges, Thomas A.; Marson, Ralph G.

    2009-01-01

    The Atacama Large Millimeter Array (ALMA) is a revolutionary millimeter and submillimeter array being developed on the Atacama plateau of northern Chile. An international partnership lead by NRAO, ESO, and NAOJ this powerful and flexible telescope will provide unprecedented observations of this relatively unexplored frequency range. The control subsystem for the Atacama Large Millimeter Array must coordinate the monitor and control of at least sixty six antennas (in four different styles), two correlators, and all of the ancillary equipment (samplers, local oscillators, front ends, etc.). This equipment will be spread over tens of kilometers and operated remotely. Operation of the array requires a robust, scalable, and maintainable real time control system. The real time control system is responsible for monitoring and control of any devices where there are fixed deadlines. Examples in the ALMA context are antenna pointing and fringe tracking. Traditionally the real time portion of a large software system is an intricate and error prone portion of the software. As a result the real time portion is very expensive in terms of effort expended both during construction and during maintenance phases of a project. The ALMA real time control system uses a Linux based real time operating system to interact with the hardware and the CORBA based ALMA Common Software to communicate in the distributed computing environment. Mixing the requirements of real time computing and the non-deterministic CORBA middleware has produced an interesting design. We discuss the architecture, design, and implementation of the ALMA real time control system. Highlight some lessons learned along the way, and justify our assertion that this should be the last large scale real time control system in radio astronomy.

  3. Set theoretical aspects of real analysis

    CERN Document Server

    Kharazishvili, Alexander B

    2014-01-01

    This book addresses a number of questions in real analysis and classical measure theory that are of a set-theoretic flavor. Accessible to graduate students, the beginning of the book presents introductory topics on real analysis and Lebesque measure theory. These topics highlight the boundary between fundamental concepts of measurability and non-measurability for point sets and functions. The remainder of the book deals with more specialized material on set-theoretical real analysis. Problems are included at the end of each chapter.

  4. Systems engineering real estate development projects

    Science.gov (United States)

    Gusakova, Elena; Titarenko, Boris; Stepanov, Vitaliy

    2017-10-01

    In recent years, real estate development has accumulated a wealth of experience in implementing major projects, which requires comprehension and systematization. The scientific instrument of system engineering is studied in the article and is substantively interpreted with reference to real estate development projects. The most perspective approaches and models are substantiated, allowing strategically to plan the life cycle of the project as a whole, and also to solve the engineering butt problems of the project. The relevance of further scientific studies of regularities and specifics of the life cycle of real estate development projects conducted at the Moscow State University of Economics and Management at the ISTA department is shown.

  5. The Cluster Enterprise of Real Property Business

    DEFF Research Database (Denmark)

    Stubkjær, Erik

    2008-01-01

    In order to apply the potential of available web technology, business processes must be described in a coherent way across organizational boundaries. Business processes include geographical data in the context of constructions, as well as environmental affairs and real property transactions, among...... others. The paper regards the domain of real property transactions. Drawing on research of European scope, it analyzes the framework of business processes in order to facilitate adoption of service oriented architectures. The paper motivates the notion of a ‘cluster enterprise' and identifies further...... steps towards a standardization of real estate business....

  6. Real-Time Java Commercial Product Assessment

    National Research Council Canada - National Science Library

    Piszcz, Alan; Vidrine, Kent

    2000-01-01

    .... Specification status and implementation towards an industry standard application-programming interface are split between two consortiums striving to introduce different paradigms of Java integration with real-time (RT) services...

  7. Studying real-world perceptual expertise.

    Science.gov (United States)

    Shen, Jianhong; Mack, Michael L; Palmeri, Thomas J

    2014-01-01

    Significant insights into visual cognition have come from studying real-world perceptual expertise. Many have previously reviewed empirical findings and theoretical developments from this work. Here we instead provide a brief perspective on approaches, considerations, and challenges to studying real-world perceptual expertise. We discuss factors like choosing to use real-world versus artificial object domains of expertise, selecting a target domain of real-world perceptual expertise, recruiting experts, evaluating their level of expertise, and experimentally testing experts in the lab and online. Throughout our perspective, we highlight expert birding (also called birdwatching) as an example, as it has been used as a target domain for over two decades in the perceptual expertise literature.

  8. Studying Real-World Perceptual Expertise

    Directory of Open Access Journals (Sweden)

    Jianhong eShen

    2014-08-01

    Full Text Available Significant insights into visual cognition have come from studying real-world perceptual expertise. Many have previously reviewed empirical findings and theoretical developments from this work. Here we instead provide a brief perspective on approaches, considerations, and challenges to studying real-world perceptual expertise. We discuss factors like choosing to use real-world versus artificial object domains of expertise, selecting a target domain of real-world perceptual expertise, recruiting experts, evaluating their level of expertise, and experimentally testing experts in the lab and online. Throughout our perspective, we highlight expert birding (also called birdwatching as an example, as it has been used as a target domain for over two decades in the perceptual expertise literature.

  9. Working with a Real Estate Developer.

    Science.gov (United States)

    Bell, Allen; Henderson, Mark

    2001-01-01

    Discusses outsourcing to an experienced real estate developer as a more efficient means of meeting school development goals on a strict time schedule. Advantages of outsourcing are covered as is advice on selecting the right development firm. (GR)

  10. The multiple reals of workplace learning

    Directory of Open Access Journals (Sweden)

    Kerry Harman

    2014-04-01

    Full Text Available The multiple reals of workplace learning are explored in this paper. Drawing on a Foucauldian conceptualisation of power as distributed, relational and productive, networks that work to produce particular objects and subjects as seemingly natural and real are examined. This approach enables different reals of workplace learning to be traced. Data from a collaborative industry-university research project is used to illustrate the approach, with a focus on the intersecting practices of a group of professional developers and a group of workplace learning researchers. The notion of multiple reals holds promise for research on workplace learning as it moves beyond a view of reality as fixed and singular to a notion of reality as performed in and through a diversity of practices, including the practices of workplace learning researchers.

  11. Lagoa Real - some alternatives of production

    International Nuclear Information System (INIS)

    Ferreira, A.B.U.

    1984-09-01

    The possibilities of uranium recovery in Lagoa Real are presented, showing that the complementary drilling must be oriented to attend the necessities of recoverable reserve and that justify the implantation of mineral-industrial complex. (author)

  12. XML does Real Programmers a Service

    Energy Technology Data Exchange (ETDEWEB)

    Gorton, Ian

    2008-09-01

    As the sun slowly sets of this first decade of the new millenium, it seems appropriate to update the sojourn of the real programmers as they adapt to their ever changing technical and business environment. Real Programmers were perfectly characterized and differentiated from their quiche-eating, Pascal programming brethren in Ed Post’s seminal “Real Programmers Don’t Use Pascal” (Datamation, 1983). My follow-up ("Real programmers do use Delphi," Software, IEEE , vol.12, no.6, pp.8, 10, 12-, Nov 1995) charted their evolution from FORTRAN-only programmers to embracing a wider range of mainstream languages and tools that still afforded ample opportunity for creativity, game-playing, irregular work hours, and importantly, long-term job security.

  13. Real Property Project Tracking System (RPPTS)

    Data.gov (United States)

    Department of Veterans Affairs — The Real Property Project Tracking System (RPPTS), formerly known as the Lease/Project Tracking (LEASE) database, contains information about lease and land projects...

  14. Transformation of Real Spherical Harmonics under Rotations

    Science.gov (United States)

    Romanowski, Z.; Krukowski, St.; Jalbout, A. F.

    2008-08-01

    The algorithm rotating the real spherical harmonics is presented. The convenient and ready to use formulae for l = 0, 1, 2, 3 are listed. The rotation in R3 space is determined by the rotation axis and the rotation angle; the Euler angles are not used. The proposed algorithm consists of three steps. (i) Express the real spherical harmonics as the linear combination of canonical polynomials. (ii) Rotate the canonical polynomials. (iii) Express the rotated canonical polynomials as the linear combination of real spherical harmonics. Since the three step procedure can be treated as a superposition of rotations, the searched rotation matrix for real spherical harmonics is a product of three matrices. The explicit formulae of matrix elements are given for l = 0, 1, 2, 3, what corresponds to s, p, d, f atomic orbitals.

  15. Reality Isn't Real Any More

    Science.gov (United States)

    Treichler, Martha

    1975-01-01

    Teachers have a responsibility to their students in providing them with the understanding that mankind sometimes has an objective hold on his world and sometimes has a subjective view of his world, thus preparing students for the real world. (RK)

  16. Visualization in Real-Time Experiment Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The objective of this project will be to migrate some of the outputs from the WFF Mission Planning Lab (MPL) into a real-time visualization system.  The MPL is...

  17. Universidades virtuales: ¿aprendizaje real?

    OpenAIRE

    Valenzuela González, Jaime R.

    2015-01-01

    1. Introducción; 2. Modalidades educativas; 3. Concepto y características de las universidades virtuales; 4. Un modelo de universidad virtual; 5. Universidades virtuales ¿aprendizaje real?; 6. Tensiones de las universidades virtuales; 7. Referencias

  18. Interactive Real-time Magnetic Resonance Imaging

    DEFF Research Database (Denmark)

    Brix, Lau

    seeks to implement and assess existing reconstruction algorithms using multi-processors of modern graphics cards and many-core computer processors and to cover some of the potential clinical applications which might benefit from using an interactive real-time MRI system. First an off......Real-time acquisition, reconstruction and interactively changing the slice position using magnetic resonance imaging (MRI) have been possible for years. However, the current clinical use of interactive real-time MRI is limited due to an inherent low spatial and temporal resolution. This PhD project......-line, but interactive, slice alignment tool was used to support the notion that 3D blood flow quantification in the heart possesses the ability to obtain curves and volumes which are not statistical different from standard 2D flow. Secondly, the feasibility of an interactive real-time MRI system was exploited...

  19. Real variables with basic metric space topology

    CERN Document Server

    Ash, Robert B

    2009-01-01

    Designed for a first course in real variables, this text presents the fundamentals for more advanced mathematical work, particularly in the areas of complex variables, measure theory, differential equations, functional analysis, and probability. Geared toward advanced undergraduate and graduate students of mathematics, it is also appropriate for students of engineering, physics, and economics who seek an understanding of real analysis.The author encourages an intuitive approach to problem solving and offers concrete examples, diagrams, and geometric or physical interpretations of results. Deta

  20. A basic course in real analysis

    CERN Document Server

    Kumar, Ajit

    2014-01-01

    Real Number SystemAlgebra of the Real Number System Upper and Lower Bounds LUB Property and Its Applications Absolute Value and Triangle Inequality Sequences and Their Convergence Sequences and Their Convergence Cauchy Sequences Monotone Sequences Sandwich Lemma Some Important Limits Sequences Diverging to Subsequences Sequences Defined Recursively Continuity Continuous Functions Definition of Continuity Intermediate Value Theorem .Extreme Value Theorem Monotone Functions Limits Uniform Continuity Continuous Extensions Differentiation Differentiability of Functions Mean Value Theorems L'Hospit

  1. Real time monitoring of electron processors

    International Nuclear Information System (INIS)

    Nablo, S.V.; Kneeland, D.R.; McLaughlin, W.L.

    1995-01-01

    A real time radiation monitor (RTRM) has been developed for monitoring the dose rate (current density) of electron beam processors. The system provides continuous monitoring of processor output, electron beam uniformity, and an independent measure of operating voltage or electron energy. In view of the device's ability to replace labor-intensive dosimetry in verification of machine performance on a real-time basis, its application to providing archival performance data for in-line processing is discussed. (author)

  2. Competitiveness and strategic flexibility through real options

    Directory of Open Access Journals (Sweden)

    SUCIU Titus

    2017-01-01

    Full Text Available The paper analyses the issue of competitiveness in the industry sector, offering an interesting theoretical framework about competitiveness and an application based on real options analysis. It addresses the theme of real options decision-making in multinational industrial companies and models an investment decision. The best strategy leads to investments in a new project even if the business environment is dynamic. The results help managers in the decision-making process, with a view to obtaining performance.

  3. Idealization of The Real Stirling Cycle

    OpenAIRE

    Červenka Libor

    2016-01-01

    The paper presents a potential idealization of the real Stirling cycle. This idealization is performed by modifying the piston movement corresponding to the ideal Stirling cycle. The focus is on the cycle thermodynamics with respect to the indicated efficiency and indicated power. A detailed 1-D simulation model of a Stirling engine is used as a tool for this assessment. The model includes real non-zero volumes of heater, regenerator, cooler and connecting pipe. The model is created in the GT...

  4. Franchise ownership redirection: real options perspective

    OpenAIRE

    Nugroho, Lukito Adi

    2016-01-01

    Background: For over 40 years, the franchise ownership redirection hypothesis has attracted the attention of many scholars. This study, differing from previous ones, proposes an alternative approach for this hypothesis using a real options framework with the extension of agency theory. Method: The real options model is built using the least square Monte Carlo method, where the franchisor's decision to franchise is perceived as a deferred investment while maintaining the right of future acquis...

  5. Scala for Real-Time Systems?

    DEFF Research Database (Denmark)

    Schoeberl, Martin

    2015-01-01

    Java served well as a general-purpose language. However, during its two decades of constant change it has gotten some weight and legacy in the language syntax and the libraries. Furthermore, Java's success for real-time systems is mediocre. Scala is a modern object-oriented and functional languag...... with interesting new features. Although a new language, it executes on a Java virtual machine, reusing that technology. This paper explores Scala as language for future real-time systems....

  6. Real-time information from seismic networks

    OpenAIRE

    Köhler, Nina

    2010-01-01

    In this thesis, the performance of a novel methodology for the real-time estimation of seismic source parameters is evaluated which can be used in earthquake early warning systems. The method, PreSEIS, uses artificial neural networks to map seismic observations from a network of sensors onto likely source parameters. The method is veryfied using two real earthquake datasets, as well as a synthetic earthquake dataset to evaluate the performance of the Istanbul early warning system.

  7. Real Estate and the Asian Crisis

    OpenAIRE

    Quigley, John M.

    2002-01-01

    This paper suggests that activities in the real estate markets in Southeast and East Asian economies were an important contributing force to the financial crises of 1997 in the Asian economies. The analysis relies upon unpublished data reported contemporaneously by financial institutions and market watchers to document the extent of the imbalances in the real property market that were evident to informed observers at the time of the financial collapse. The analysis argues that a series of ref...

  8. Why Are Real Interest Rates So High?

    OpenAIRE

    Zvi Bodie; Alex Kane; Robert L. McDonald

    1983-01-01

    This paper applies the Capital Asset Pricing Model to help explain the anomalous behavior of real interest rates during the last several years. Specifically,we are able to show that the increased volatility of bond prices since the change in Federal Reserve operating procedure in October 1979 has substantially increased the required real risk premium on long term bonds. We also consider and reject the possibility that increased risk alone accounts for the recent increase in the short-term rea...

  9. Real Exchange Rates in Advanced Transition Economies

    Directory of Open Access Journals (Sweden)

    Sanja Grubacic

    2015-10-01

    Full Text Available The recent evidence from Eastern Europe suggests that one of the major obstacles towards the adoption of euro may lie in the impact that the recession of 2008 exerted on the trajectory of real exchange rates in new member countries (European Commission, 2015.  This paper aims to establish and explain the relationship between the external shocks derived from the global financial crisis and recession of 2008 and equilibrium real exchange rate in advanced transition economies of Eastern Europe. The interplay between the external and internal balances is explained by developing an inter-temporal optimizing model of the real exchange rate determination in a small open economy with structural distortions. The results of our model suggest that, in the aftermath of recession, if the Eastern European economies attempt to restore and maintain the balance between the consumption, saving, and investment, the equilibrium real exchange rate will tend to reverse its trajectory from appreciation to depreciation over time in order to encourage a greater production in the future. The equilibrium real exchange rate depreciation in the future may obtain either as a result of an increase in the direct subsidies on investment or as a result of reduced subsidies on the "net-of-investment" income.  The deprecation of countries’ real exchange rate, however, may continue to act as an effective constraint against the adoption of euro.

  10. 33 CFR 211.1 - Real estate defined.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Real estate defined. 211.1... DEFENSE REAL ESTATE ACTIVITIES OF THE CORPS OF ENGINEERS IN CONNECTION WITH CIVIL WORKS PROJECTS Real Estate; General § 211.1 Real estate defined. The term real estate as used in this part includes land...

  11. Real-time RT-PCR analysis of human histidine decarboxylase, a new marker for several types of leukemia and cancer.

    Science.gov (United States)

    Melgarejo, Esther; Medina, Miguel Angel; Paz, José Carlos; Sánchez-Jiménez, Francisca; Urdiales, José Luis

    2006-01-01

    Histamine is involved in different physiological and pathological responses, such as immune response, gastric acid secretion or neurotransmission, as either angiogenesis or cancer. Histidine decarboxylase (HDC) catalyzes the formation of histamine from histidine. HDC has been suggested as a new marker for neuroendocrine differentiation, inflammatory pathologies and several leukemia and highly malignant forms of cancer, such as melanoma and small cell lung carcinoma. In the present work, we describe the use of Syber Green-based quantitative real-time RT-PCR to determine the expression of histidine decarboxylase in human cells and tissue. As an internal control, glyceraldehyde 3-phosphate dehydrogenase was also amplified. The linear dynamic range of the assay covered 4 orders of magnitude for HDC amplification. The detection limit was 0.1 ng of total RNA extracted from HMC-1 cells. This method is simple, rapid, sensitive, and quantitative, and allows for the specific identification of cells and tissue expressing HDC, stressing its potential diagnostic usefulness in malignancies in which HDC is described as a new marker.

  12. TRERC-TEA [Texas Real Estate Research Center-Texas Education Agency] Real Estate Curriculum Workshop Committee Report.

    Science.gov (United States)

    Lyon, Robert

    The document contains a summary report of a community college real estate teachers' workshop organized to develop course outlines for the various areas in the real estate curriculum. Curriculum outlines are presented, with varying degrees of detail included, for the following eight subjects: real estate appraisal; real estate brokage; real estate…

  13. REAL TIME SYSTEM OPERATIONS 2006-2007

    Energy Technology Data Exchange (ETDEWEB)

    Eto, Joseph H.; Parashar, Manu; Lewis, Nancy Jo

    2008-08-15

    The Real Time System Operations (RTSO) 2006-2007 project focused on two parallel technical tasks: (1) Real-Time Applications of Phasors for Monitoring, Alarming and Control; and (2) Real-Time Voltage Security Assessment (RTVSA) Prototype Tool. The overall goal of the phasor applications project was to accelerate adoption and foster greater use of new, more accurate, time-synchronized phasor measurements by conducting research and prototyping applications on California ISO's phasor platform - Real-Time Dynamics Monitoring System (RTDMS) -- that provide previously unavailable information on the dynamic stability of the grid. Feasibility assessment studies were conducted on potential application of this technology for small-signal stability monitoring, validating/improving existing stability nomograms, conducting frequency response analysis, and obtaining real-time sensitivity information on key metrics to assess grid stress. Based on study findings, prototype applications for real-time visualization and alarming, small-signal stability monitoring, measurement based sensitivity analysis and frequency response assessment were developed, factory- and field-tested at the California ISO and at BPA. The goal of the RTVSA project was to provide California ISO with a prototype voltage security assessment tool that runs in real time within California ISO?s new reliability and congestion management system. CERTS conducted a technical assessment of appropriate algorithms, developed a prototype incorporating state-of-art algorithms (such as the continuation power flow, direct method, boundary orbiting method, and hyperplanes) into a framework most suitable for an operations environment. Based on study findings, a functional specification was prepared, which the California ISO has since used to procure a production-quality tool that is now a part of a suite of advanced computational tools that is used by California ISO for reliability and congestion management.

  14. Real-Time Polymerase Chain Reaction (PCR) Capability in Space

    Data.gov (United States)

    National Aeronautics and Space Administration — The goal of this project is enabling the real-time polymerase chain reaction (real-time PCR) technology in space. In space, the real-time PCR technology can be used...

  15. Medical real estate in an age of reform.

    Science.gov (United States)

    Hammond, Laca Wong; Camp, Philip J

    2011-04-01

    The following are four ways healthcare organizations are fulfilling their medical real estate needs in an era of change: Real estate monetization. Renovation of existing facilities. A careful focus on containing materials costs. Joint ventures with real estate organizations.

  16. Network protocols for real-time applications

    Science.gov (United States)

    Johnson, Marjory J.

    1987-01-01

    The Fiber Distributed Data Interface (FDDI) and the SAE AE-9B High Speed Ring Bus (HSRB) are emerging standards for high-performance token ring local area networks. FDDI was designed to be a general-purpose high-performance network. HSRB was designed specifically for military real-time applications. A workshop was conducted at NASA Ames Research Center in January, 1987 to compare and contrast these protocols with respect to their ability to support real-time applications. This report summarizes workshop presentations and includes an independent comparison of the two protocols. A conclusion reached at the workshop was that current protocols for the upper layers of the Open Systems Interconnection (OSI) network model are inadequate for real-time applications.

  17. Real depletion in nodal diffusion codes

    International Nuclear Information System (INIS)

    Petkov, P.T.

    2002-01-01

    The fuel depletion is described by more than one hundred fuel isotopes in the advanced lattice codes like HELIOS, but only a few fuel isotopes are accounted for even in the advanced steady-state diffusion codes. The general assumption that the number densities of the majority of the fuel isotopes depend only on the fuel burnup is seriously in error if high burnup is considered. The real depletion conditions in the reactor core differ from the asymptotic ones at the stage of lattice depletion calculations. This study reveals which fuel isotopes should be explicitly accounted for in the diffusion codes in order to predict adequately the real depletion effects in the core. A somewhat strange conclusion is that if the real number densities of the main fissionable isotopes are not explicitly accounted for in the diffusion code, then Sm-149 should not be accounted for either, because the net error in k-inf is smaller (Authors)

  18. Studying Complex Interactions in Real Time

    DEFF Research Database (Denmark)

    Mønster, Dan

    2017-01-01

    The study of human behavior must take into account the social context, and real-time, networked experiments with multiple participants is one increasingly popular way to achieve this. In this paper a framework based on Python and XMPP is presented that aims to make it easy to develop such behavio......The study of human behavior must take into account the social context, and real-time, networked experiments with multiple participants is one increasingly popular way to achieve this. In this paper a framework based on Python and XMPP is presented that aims to make it easy to develop...... such behavioral experiments. An illustrative example of how the framework can be used is also presented. This example is a real experiment, which is currently gathering data in the lab....

  19. Failure analysis of real-time systems

    International Nuclear Information System (INIS)

    Jalashgar, A.; Stoelen, K.

    1998-01-01

    This paper highlights essential aspects of real-time software systems that are strongly related to the failures and their course of propagation. The significant influence of means-oriented and goal-oriented system views in the description, understanding and analysing of those aspects is elaborated. The importance of performing failure analysis prior to reliability analysis of real-time systems is equally addressed. Problems of software reliability growth models taking the properties of such systems into account are discussed. Finally, the paper presents a preliminary study of a goal-oriented approach to model the static and dynamic characteristics of real-time systems, so that the corresponding analysis can be based on a more descriptive and informative picture of failures, their effects and the possibility of their occurrence. (author)

  20. Real Time Structured Light and Applications

    DEFF Research Database (Denmark)

    Wilm, Jakob

    ]. A high performance flexible open source software toolkit is presented [Contribution C], which makes real time scanning possible on commodity hardware. Further, an approach is presented to correct for motion artifacts in dynamic scenes [Contribution E]. An application for such systems is presented......, increased processing power, and methods presented in this thesis, it is possible to perform structured light scans in real time with 20 depth measurements per second. This offers new opportunities for studying dynamic scenes, quality control, human-computer interaction and more. This thesis discusses...... several aspects of real time structured light systems and presents contributions within calibration, scene coding and motion correction aspects. The problem of reliable and fast calibration of such systems is addressed with a novel calibration scheme utilising radial basis functions [Contribution B...

  1. Tumor necrosis factor alpha increases epithelial barrier permeability by disrupting tight junctions in Caco-2 cells.

    Science.gov (United States)

    Cui, W; Li, L X; Sun, C M; Wen, Y; Zhou, Y; Dong, Y L; Liu, P

    2010-04-01

    The objectives of this study were to determine the effect of tumor necrosis factor alpha (TNF-alpha) on intestinal epithelial cell permeability and the expression of tight junction proteins. Caco-2 cells were plated onto Transwell microporous filters and treated with TNF-alpha (10 or 100 ng/mL) for 0, 4, 8, 16, or 24 h. The transepithelial electrical resistance and the mucosal-to-serosal flux rates of the established paracellular marker Lucifer yellow were measured in filter-grown monolayers of Caco-2 intestinal cells. The localization and expression of the tight junction protein occludin were detected by immunofluorescence and Western blot analysis, respectively. SYBR-Green-based real-time PCR was used to measure the expression of occludin mRNA. TNF-alpha treatment produced concentration- and time-dependent decreases in Caco-2 transepithelial resistance and increases in transepithelial permeability to the paracellular marker Lucifer yellow. Western blot results indicated that TNF-alpha decreased the expression of phosphorylated occludin in detergent-insoluble fractions but did not affect the expression of non-phosphorylated occludin protein. Real-time RT-PCR data showed that TNF-alpha did not affect the expression of occludin mRNA. Taken together, our data demonstrate that TNF-alpha increases Caco-2 monolayer permeability, decreases occludin protein expression and disturbs intercellular junctions.

  2. Tumor necrosis factor alpha increases epithelial barrier permeability by disrupting tight junctions in Caco-2 cells

    Directory of Open Access Journals (Sweden)

    W. Cui

    2010-04-01

    Full Text Available The objectives of this study were to determine the effect of tumor necrosis factor alpha (TNF-α on intestinal epithelial cell permeability and the expression of tight junction proteins. Caco-2 cells were plated onto Transwell® microporous filters and treated with TNF-α (10 or 100 ng/mL for 0, 4, 8, 16, or 24 h. The transepithelial electrical resistance and the mucosal-to-serosal flux rates of the established paracellular marker Lucifer yellow were measured in filter-grown monolayers of Caco-2 intestinal cells. The localization and expression of the tight junction protein occludin were detected by immunofluorescence and Western blot analysis, respectively. SYBR-Green-based real-time PCR was used to measure the expression of occludin mRNA. TNF-α treatment produced concentration- and time-dependent decreases in Caco-2 transepithelial resistance and increases in transepithelial permeability to the paracellular marker Lucifer yellow. Western blot results indicated that TNF-α decreased the expression of phosphorylated occludin in detergent-insoluble fractions but did not affect the expression of non-phosphorylated occludin protein. Real-time RT-PCR data showed that TNF-α did not affect the expression of occludin mRNA. Taken together, our data demonstrate that TNF-α increases Caco-2 monolayer permeability, decreases occludin protein expression and disturbs intercellular junctions.

  3. When only the real thing will do: junior medical students’ learning from real patients

    NARCIS (Netherlands)

    Bell, Kathryn; Boshuizen, Els; Scherpbier, Albert; Dornan, Tim

    2010-01-01

    Bell, K., Boshuizen, H. P. A., Scherpbier, A. J. J. A., & Dornan, T. L. (2009). When only the real thing will do: junior medical students' learning from real patients. Medical Education, 43(11), 1036-1043. doi:10.1111/j.1365-2923.2009.03508.x

  4. Sensemaking of real estate management using real options and scenario planning

    NARCIS (Netherlands)

    van Reedt Dortland, Maartje; Voordijk, Johannes T.; Dewulf, Geert P.M.R.

    2012-01-01

    Healthcare across the world is facing many uncertainties. In Dutch healthcare, a recent policy change forces health organisations to deal more efficiently with real estate which makes flexibility more necessary. In order to support real estate managers in decision making in flexibility, we developed

  5. Scalar dark matter: real vs complex

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Hongyan; Zheng, Sibo [Department of Physics, Chongqing University,Chongqing 401331 (China)

    2017-03-27

    We update the parameter spaces for both a real and complex scalar dark matter via the Higgs portal. In the light of constraints arising from the LUX 2016 data, the latest Higgs invisible decay and the gamma ray spectrum, the dark matter resonant mass region is further restricted to a narrow window between 54.9−62.3 GeV in both cases, and its large mass region is excluded until 834 GeV and 3473 GeV for the real and complex scalar, respectively.

  6. Real Time Linux - The RTOS for Astronomy?

    Science.gov (United States)

    Daly, P. N.

    The BoF was attended by about 30 participants and a free CD of real time Linux-based upon RedHat 5.2-was available. There was a detailed presentation on the nature of real time Linux and the variants for hard real time: New Mexico Tech's RTL and DIAPM's RTAI. Comparison tables between standard Linux and real time Linux responses to time interval generation and interrupt response latency were presented (see elsewhere in these proceedings). The present recommendations are to use RTL for UP machines running the 2.0.x kernels and RTAI for SMP machines running the 2.2.x kernel. Support, both academically and commercially, is available. Some known limitations were presented and the solutions reported e.g., debugging and hardware support. The features of RTAI (scheduler, fifos, shared memory, semaphores, message queues and RPCs) were described. Typical performance statistics were presented: Pentium-based oneshot tasks running > 30kHz, 486-based oneshot tasks running at ~ 10 kHz, periodic timer tasks running in excess of 90 kHz with average zero jitter peaking to ~ 13 mus (UP) and ~ 30 mus (SMP). Some detail on kernel module programming, including coding examples, were presented showing a typical data acquisition system generating simulated (random) data writing to a shared memory buffer and a fifo buffer to communicate between real time Linux and user space. All coding examples were complete and tested under RTAI v0.6 and the 2.2.12 kernel. Finally, arguments were raised in support of real time Linux: it's open source, free under GPL, enables rapid prototyping, has good support and the ability to have a fully functioning workstation capable of co-existing hard real time performance. The counter weight-the negatives-of lack of platforms (x86 and PowerPC only at present), lack of board support, promiscuous root access and the danger of ignorance of real time programming issues were also discussed. See ftp://orion.tuc.noao.edu/pub/pnd/rtlbof.tgz for the StarOffice overheads

  7. Recent Advances in Real Options Signaling Games

    Directory of Open Access Journals (Sweden)

    George Rigopoulos

    2014-12-01

    Full Text Available Financial economics have benefited from game theory advances and its application to various domains where competition is present. Following relevant research this paper presents a literature review of recent works on the intersection of real options and game theory, especially on signaling games. A preliminary background is presented initially and next a review of the works is presented. The paper compiles existing works to a useful reference for researchers in the field. Keywords: Signaling games; Capital budgeting, Real options, Game theory, Discrete time games

  8. Real and étale cohomology

    CERN Document Server

    Scheiderer, Claus

    1994-01-01

    This book makes a systematic study of the relations between the étale cohomology of a scheme and the orderings of its residue fields. A major result is that in high degrees, étale cohomology is cohomology of the real spectrum. It also contains new contributions in group cohomology and in topos theory. It is of interest to graduate students and researchers who work in algebraic geometry (not only real) and have some familiarity with the basics of étale cohomology and Grothendieck sites. Independently, it is of interest to people working in the cohomology theory of groups or in topos theory.

  9. Real-time systems scheduling 2 focuses

    CERN Document Server

    Chetto, Maryline

    2014-01-01

    Real-time systems are used in a wide range of applications, including control, sensing, multimedia, etc. Scheduling is a central problem for these computing/communication systems since it is responsible for software execution in a timely manner. This book, the second of two volumes on the subject, brings together knowledge on specific topics and discusses the recent advances for some of them.  It addresses foundations as well as the latest advances and findings in real-time scheduling, giving comprehensive references to important papers, but the chapters are short and not overloaded with co

  10. Machine vision for real time orbital operations

    Science.gov (United States)

    Vinz, Frank L.

    1988-01-01

    Machine vision for automation and robotic operation of Space Station era systems has the potential for increasing the efficiency of orbital servicing, repair, assembly and docking tasks. A machine vision research project is described in which a TV camera is used for inputing visual data to a computer so that image processing may be achieved for real time control of these orbital operations. A technique has resulted from this research which reduces computer memory requirements and greatly increases typical computational speed such that it has the potential for development into a real time orbital machine vision system. This technique is called AI BOSS (Analysis of Images by Box Scan and Syntax).

  11. SignalR real time application development

    CERN Document Server

    Ingebrigtsen, Einar

    2013-01-01

    This step-by-step guide gives you practical advice, tips, and tricks that will have you writing real-time apps quickly and easily.If you are a .NET developer who wants to be at the cutting edge of development, then this book is for you. Real-time application development is made simple in this guide, so as long as you have basic knowledge of .NET, a copy of Visual Studio, and NuGet installed, you are ready to go.

  12. Let's just hold a real vote.

    Science.gov (United States)

    Phillips, Gareth

    2017-04-12

    The planned RCN consultation to gauge how nurses feel about balloting on industrial action is a step in the right direction, but for heaven's sake why don't we just hold a real vote? No one is interested in a test vote - not the media, not members. A real vote would be headline news and RCN members would be enthused and motivated. As it is, we are just kicking the ball into the long grass yet again. There will be a low turnout and we will end up shrugging our shoulders and saying 'There's nothing to be done'. Jeremy Hunt must be laughing his socks off.

  13. The reals as rational Cauchy filters

    OpenAIRE

    Weiss, Ittay

    2015-01-01

    We present a detailed and elementary construction of the real numbers from the rational numbers a la Bourbaki. The real numbers are defined to be the set of all minimal Cauchy filters in $\\mathbb{Q}$ (where the Cauchy condition is defined in terms of the absolute value function on $\\mathbb{Q}$) and are proven directly, without employing any of the techniques of uniform spaces, to form a complete ordered field. The construction can be seen as a variant of Bachmann's construction by means of ne...

  14. Real-time systems scheduling fundamentals

    CERN Document Server

    Chetto, Maryline

    2014-01-01

    Real-time systems are used in a wide range of applications, including control, sensing, multimedia, etc.  Scheduling is a central problem for these computing/communication systems since responsible of software execution in a timely manner. This book provides state of knowledge in this domain with special emphasis on the key results obtained within the last decade. This book addresses foundations as well as the latest advances and findings in Real-Time Scheduling, giving all references to important papers. But nevertheless the chapters will be short and not overloaded with confusing details.

  15. Image processing in real time radiography

    International Nuclear Information System (INIS)

    Link, R.; Nuding, W.; Sauevwein, K.; Souw, E.K.

    1985-01-01

    Image processing in real time radiography has become an important feature to improve the detectibility of defects. However, often enough, impressed by the tremendous success of image processing of e.g. evaluation of Landsat pictures, people expect the same or nearly the same effect in NDT applications. The magic word image processing thus results in unrealistic demands to the capability even of highly sophisticated image processing systems. In this paper the possibilities as well as the different tasks of image processing in the field of real time radiography is discussed

  16. Real-Time Mesoscale Prediction on workstations.

    Science.gov (United States)

    Cotton, William R.; Thompson, Gregory; Mieike, Paul W., Jr.

    1994-03-01

    Experience in performing real-time mesoscale numerical prediction forecasts using the Regional Atmospheric Modeling System (RAMS) over Colorado for a winter season on high-performance workstations is summarized. Performance evaluation is done for specific case studies and, statistically, for the entire winter season. RAMS forecasts are also compared with nested grid model forecasts. In addition, RAMS precipitation forecasts with a simple "dump bucket" scheme are compared with explicit, bulk microphysics parameterization schemes. The potential applications and political/ social problems of having a readily accessible, real-time mesoscale forecasting capability on low-cost, high-performance workstations is discussed.

  17. Euro Membership as a Real Option Trigger

    DEFF Research Database (Denmark)

    Aabo, Tom; Pantzalis, Christos

    2007-01-01

    -adopters (UK, Sweden, and Denmark) to exercise various forms of real options such as to establish alliances / partnerships, to enter new markets / market segments, to switch suppliers, and to generally expand in the Euro-area. The study furthermore shows that small, profitable and financially constrained firms...... are particularly likely to exercise such real options triggered by the introduction of the Euro. The results go beyond the immediate trade effects, which empirical studies have shown to be weak and without trade diversion as to the three non-adopters, and provide important insights about the potential long...

  18. Aerial pruning mechanism, initial real environment test.

    Science.gov (United States)

    Molina, Javier; Hirai, Shinichi

    2017-01-01

    In this research, a pruning mechanism for aerial pruning tasks is tested in a real environment. Since the final goal of the aerial pruning robot will be to prune tree branches close to power lines, some experiments related to wireless communication and pruning performance were conducted. The experiments consisted of testing the communication between two XBee RF modules for monitoring purposes as well as testing the speed control of the circular saw used for pruning tree branches. Results show that both the monitoring and the pruning tasks were successfully done in a real environment.

  19. Collecting data in real time with postcards

    DEFF Research Database (Denmark)

    Yee, Kwang Chien; Kanstrup, Anne Marie; Bertelsen, Pernille

    2013-01-01

    Systems. These methods often involve cross-sectional, retrospective data collection. This paper describes the postcard method for prospective real-time data collection, both in paper format and electronic format. This paper then describes the results obtained using postcard techniques in Denmark...... and Australia. The benefits of this technique are illustrated. There are limitations in using postcard techniques and this paper provides a detail discussion about these limitations. Postcard techniques provide unique advantages in understanding real time healthcare context and it is an important technique...

  20. Real analysis and probability solutions to problems

    CERN Document Server

    Ash, Robert P

    1972-01-01

    Real Analysis and Probability: Solutions to Problems presents solutions to problems in real analysis and probability. Topics covered range from measure and integration theory to functional analysis and basic concepts of probability; the interplay between measure theory and topology; conditional probability and expectation; the central limit theorem; and strong laws of large numbers in terms of martingale theory.Comprised of eight chapters, this volume begins with problems and solutions for the theory of measure and integration, followed by various applications of the basic integration theory.

  1. Real analysis modern techniques and their applications

    CERN Document Server

    Folland, Gerald B

    1999-01-01

    An in-depth look at real analysis and its applications-now expanded and revised.This new edition of the widely used analysis book continues to cover real analysis in greater detail and at a more advanced level than most books on the subject. Encompassing several subjects that underlie much of modern analysis, the book focuses on measure and integration theory, point set topology, and the basics of functional analysis. It illustrates the use of the general theories and introduces readers to other branches of analysis such as Fourier analysis, distribution theory, and probability theory.This edi

  2. Using Genetic Algorithms for Real Estate Appraisals

    Directory of Open Access Journals (Sweden)

    Vincenzo Del Giudice

    2017-03-01

    Full Text Available The main aim of this paper is the interpretation of the existing relationship between real estate rental prices and geographical location of housing units in a central urban area of Naples (Santa Lucia and Riviera of Chiaia neighborhoods. Genetic algorithms (GA are used for this purpose. Also, to verify the reliability of genetic algorithms for real estate appraisals and, at the same time, to show the forecasting potentialities of these techniques in the analysis of housing markets, a multiple regression analysis (MRA was applied comparing results of GA and MRA.

  3. Real economic activity and accounting information in Spanish construction and real estate firms

    Directory of Open Access Journals (Sweden)

    Juan Carlos Navarro-García

    2016-01-01

    Full Text Available The aim of this work is to determine whether the accounting information provided by construction and real estate firms in Spain reflects the real economic activity, during upswings and downturns. A relationship is found between economic conditions and financial reporting in a continental European country such as Spain. Although there is a relationship between real housing price growth and financial accounting measures, real estate companies seem to show an artificial position, since they only present a relationship with Return on Assets (ROA based on low quality income (accruals. Therefore, a different financial position influences a different behaviour in these two similar sectors. In fact, some signs alerting of possible risk in real estate firms are shown.

  4. A model of real estate and psychological factors in decision-making to buy real estate

    Directory of Open Access Journals (Sweden)

    Bojan Grum

    2015-06-01

    Full Text Available This article explores the psychological characteristics of potential real estate buyers connected with their decision to buy. Through a review of research, it reveals that most studies of psychological factors in the decision to buy real estate have a partial and dispersed orientation, and examine individual factors independently. It appears that the research area is lacking clearly defined models of psychological factors in the decision to buy real estate that would integrally and relationally explain the role of psychological characteristics of real estate buyers and their expectations in relation to a decision to buy. The article identifies two sets of psychological factors, motivational and emotional, determines their interaction with potential buyers’ expectations when deciding to purchase real estate and offers starting points for forming a model.

  5. Comprehensive and Rapid Real-Time PCR Analysis of 21 Foodborne Outbreaks

    Directory of Open Access Journals (Sweden)

    Hiroshi Fukushima

    2009-01-01

    Full Text Available A set of four duplex SYBR Green I PCR (SG-PCR assay combined with DNA extraction using QIAamp DNA Stool Mini kit was evaluated for the detection of foodborne bacteria from 21 foodborne outbreaks. The causative pathogens were detected in almost all cases in 2 hours or less. The first run was for the detection of 8 main foodborne pathogens in 5 stool specimens within 2 hours and the second run was for the detection of other unusual suspect pathogens within a further 45 minutes. After 2 to 4 days, the causative agents were isolated and identified. The results proved that for comprehensive and rapid molecular diagnosis in foodborne outbreaks, Duplex SG-PCR assay is not only very useful, but is also economically viable for one-step differentiation of causative pathogens in fecal specimens obtained from symptomatic patients. This then allows for effective diagnosis and management of foodborne outbreaks.

  6. 22 CFR 518.32 - Real property.

    Science.gov (United States)

    2010-04-01

    ... 22 Foreign Relations 2 2010-04-01 2010-04-01 true Real property. 518.32 Section 518.32 Foreign... current fair market value of the property attributable to the Federal participation in the project. (2... agency and pay the Federal Government for that percentage of the current fair market value of the...

  7. Refactoring Real-Time Java Profiles

    DEFF Research Database (Denmark)

    Søndergaard, Hans; Thomsen, Bent; Ravn, Anders P.

    2011-01-01

    Just like other software, Java profiles benefits from refactoring when they have been used and have evolved for some time. This paper presents a refactoring of the Real-Time Specification for Java (RTSJ) and the Safety Critical Java (SCJ) profile (JSR-302). It highlights core concepts and makes...

  8. Real moments of the restrictive factor

    Indian Academy of Sciences (India)

    ... Discussion Meetings · Public Lectures · Lecture Workshops · Refresher Courses · Symposia · Live Streaming. Home; Journals; Proceedings – Mathematical Sciences; Volume 119; Issue 4. Real Moments of the Restrictive Factor. Andrew Ledoan Alexandru Zaharescu. Volume 119 Issue 4 September 2009 pp 559-566 ...

  9. A foundation for real recursive function theory

    NARCIS (Netherlands)

    J.F. Costa; B. S. Loff Barreto (Bruno Serra); J. Mycka

    2009-01-01

    htmlabstractThe class of recursive functions over the reals, denoted by REC(R), was introduced by Cristopher Moore in his seminal paper written in 1995. Since then many subsequent investigations brought new results: the class REC(R) was put in relation with the class of functions generated by the

  10. Research into real-option evaluation method

    International Nuclear Information System (INIS)

    Shiba, Tsuyoshi; Wakamatsu, Hitoshi

    2002-03-01

    As an evaluational method for valuation of a corporation, an investment project, a research and development, or the evaluation technique of an enterprise strategy, a real option analysis attracts attention instead of conventional Discount Cash Flow method. The reason is that it can divert the technique for the option valuation in financial engineering to the decision-making process performed according to change in investment environment. Related references, the analysis tools, the application examples, etc. were investigated about the decision-making technique using real option analysis, and this investigation considered the application method to decision-making of the research and development at Japan Nuclear Cycle Development Institute. Consequently, since the feature is in real option analysis being the evaluation technique on condition of that business conditions and business itself also change, the real option analysis fits for evaluation of a research and development that business conditions were opaque and it turns out that the businesses are highly flexible. Moreover, it turns out that it fits also for evaluation of a capital concentration type investment issue like power plants. (author)

  11. A monotonicity conjecture for real cubic maps

    Energy Technology Data Exchange (ETDEWEB)

    Dawson, S.P. [Los Alamos National Lab., NM (United States); Galeeva, R. [Northwestern Univ., Evanston, IL (United States); Milnor, J. [State Univ. of New York, Stony Brook, NY (United States); Tresser, C. [International Business Machines Corp., Yorktown Heights, NY (United States)

    1993-12-01

    This will be an outline of work in progress. We study the conjecture that the topological entropy of a real cubic map depends ``monotonely`` on its parameters, in the sense that each locus of constant entropy in parameter space is a connected set. This material will be presented in more detail in a later paper.

  12. Lagoa Real design - Use of Acre910

    International Nuclear Information System (INIS)

    Anacleto, R.

    1982-01-01

    Some works made by ASAV.PM (Evaluation Commission for Mineral Resources, NUCLEBRAS) with computer resources and ACRE910 program, including geological interpretation of AN-03, AN-06 and AN-13 anomaly from Lagoa Real Design (Brazilian Uranium Basin), aiming discuss the space position of uranium mineralized bodies and put at disposal the computer facilities for the users are presented. (author)

  13. Real-Time Polymerase Chain Reaction

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 13; Issue 4. Real-Time Polymerase Chain Reaction - A Revolution in Diagnostics. Simarjot Singh Pabla Sarabjot Singh Pabla. General Article Volume 13 Issue 4 April 2008 pp 369-377 ...

  14. Real Time Grid Reliability Management 2005

    Energy Technology Data Exchange (ETDEWEB)

    Eto, Joe; Eto, Joe; Lesieutre, Bernard; Lewis, Nancy Jo; Parashar, Manu

    2008-07-07

    The increased need to manage California?s electricity grid in real time is a result of the ongoing transition from a system operated by vertically-integrated utilities serving native loads to one operated by an independent system operator supporting competitive energy markets. During this transition period, the traditional approach to reliability management -- construction of new transmission lines -- has not been pursued due to unresolved issues related to the financing and recovery of transmission project costs. In the absence of investments in new transmission infrastructure, the best strategy for managing reliability is to equip system operators with better real-time information about actual operating margins so that they can better understand and manage the risk of operating closer to the edge. A companion strategy is to address known deficiencies in offline modeling tools that are needed to ground the use of improved real-time tools. This project: (1) developed and conducted first-ever demonstrations of two prototype real-time software tools for voltage security assessment and phasor monitoring; and (2) prepared a scoping study on improving load and generator response models. Additional funding through two separate subsequent work authorizations has already been provided to build upon the work initiated in this project.

  15. Oversampling of wavelet frames for real dilations

    DEFF Research Database (Denmark)

    Bownik, Marcin; Lemvig, Jakob

    2012-01-01

    We generalize the Second Oversampling Theorem for wavelet frames and dual wavelet frames from the setting of integer dilations to real dilations. We also study the relationship between dilation matrix oversampling of semi-orthogonal Parseval wavelet frames and the additional shift invariance gain...

  16. Real Capital: Complementarity and Structural Characteristics

    Directory of Open Access Journals (Sweden)

    Miriasov Iurii О.

    2016-05-01

    Full Text Available Two opposite approaches to the concept of real capital are considered. The presupposition for studying capital within the neoclassical school, which can be characterized as “empirical realism”, is the homogeneity of capital. This allows to apply formal methods of studying and modeling industrial processes but excludes from consideration the properties and characteristics of capital as a real factor of production. The approach of the Austrian school corresponding to traditions of “critical realism” in the economy focuses on real properties of capital and its role in the process of public production. The aim of this study is to clarify the idea of real capital, its properties, structural characteristics and mechanisms of formation. According to this, the characteristic of two different approaches is given. It is shown that the heterogeneity of capital is the basis of complementarity of capital goods and their combinations in each particular case of their application. The same capital goods have different performance depending on the method of their combination as a result of the production plans of entrepreneurs. Emergent properties of capital are also the result of the combination. Differences in the use and combination of capital determine its overall structure, subsystems of which are natural objects at different levels included in the production, capital goods themselves and their various combinations, social capital. Each of these subsystems is relatively independent and forms a higher level subsystem.

  17. Get Real: Augmented Reality for the Classroom

    Science.gov (United States)

    Mitchell, Rebecca; DeBay, Dennis

    2012-01-01

    Kids love augmented reality (AR) simulations because they are like real-life video games. AR simulations allow students to learn content while collaborating face to face and interacting with a multimedia-enhanced version of the world around them. Although the technology may seem advanced, AR software makes it easy to develop content-based…

  18. Symbolic Victimization and Real World Fear.

    Science.gov (United States)

    Morgan, Michael

    1983-01-01

    Examines the relationship between victimization of characters in television drama and susceptibility to the viewers' cultivation of a sense of personal risk in the real world. Found that viewers whose fictional counterparts are more likely to be shown as victims show stronger associations between viewing and perceived vulnerability. (PD)

  19. Compiling models into real-time systems

    International Nuclear Information System (INIS)

    Dormoy, J.L.; Cherriaux, F.; Ancelin, J.

    1992-08-01

    This paper presents an architecture for building real-time systems from models, and model-compiling techniques. This has been applied for building a real-time model-based monitoring system for nuclear plants, called KSE, which is currently being used in two plants in France. We describe how we used various artificial intelligence techniques for building it: a model-based approach, a logical model of its operation, a declarative implementation of these models, and original knowledge-compiling techniques for automatically generating the real-time expert system from those models. Some of those techniques have just been borrowed from the literature, but we had to modify or invent other techniques which simply did not exist. We also discuss two important problems, which are often underestimated in the artificial intelligence literature: size, and errors. Our architecture, which could be used in other applications, combines the advantages of the model-based approach with the efficiency requirements of real-time applications, while in general model-based approaches present serious drawbacks on this point

  20. 29 CFR 95.32 - Real property.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 1 2010-07-01 2010-07-01 true Real property. 95.32 Section 95.32 Labor Office of the Secretary of Labor GRANTS AND AGREEMENTS WITH INSTITUTIONS OF HIGHER EDUCATION, HOSPITALS, AND OTHER NON... Government after it compensates the Federal Government for that percentage of the current fair market value...

  1. Managing Uncertainty in the Real World

    Indian Academy of Sciences (India)

    Managing Uncertainty in the Real World. 2. Fuzzy Systems. Satish Kumar. In Part 1 of this article we understood the idea of a fuzzy set. We also reviewed simple operations such as fuzzy set i..~tersection, union, and complementation, and related these to geometry. In Part 2, we extend the idea offuzzy sets to explain how we ...

  2. Students Dig for Real School Gardens

    Science.gov (United States)

    Reeves, Lacey; Emeagwali, N. Susan

    2010-01-01

    There's a lot of talk about saving the environment and going green these days. But the challenge is to turn the words into action, and that includes getting young students to become part of the discussion about sustainability. The Texas-based Rainwater Environmental Alliance for Learning (REAL) School Gardens is cultivating success by providing…

  3. Managing Uncertainty in the Real World

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 4; Issue 4. Managing Uncertainty in the Real World - Fuzzy Systems. Satish Kumar. General Article ... Author Affiliations. Satish Kumar1. Department of Physics and Computer Science, Dayalbagh Educational Institute Dayalbagh, Agra 282005, India.

  4. On a Class of Real Integrals

    Science.gov (United States)

    Capelas de Oliveira, E.; Rosa, M. A. F.; Vaz, J., Jr.

    2009-01-01

    We present a calculation involving a wide class of real integrals by means of integration in the complex plane. Some particular cases where Euler numbers and Bell polynomials appear are discussed and a generalisation of some previous results is also provided. (Contains 1 table and 1 figure.)

  5. Acquisitions and Real Options : The Greenfield Alternative

    NARCIS (Netherlands)

    Brouthers, Keith D.; Dikova, Desislava

    P>Although acquisitions are a popular way to enter new markets, empirical evidence tends to indicate few benefits accrue to acquiring firms. This might be the case because firms use acquisitions when they should be employing an alternative mode of expansion. Applying real options theory to this

  6. Teaching "Real Utopias" through Experiential Learning

    Science.gov (United States)

    Peterson, Lindsey; Witt, Joseph; Huntington, Carolyn

    2015-01-01

    In this article, we describe a way to encourage students to envision "real utopias" through the Global Village experience at the Heifer Ranch in Arkansas. The Global Village experience introduces participants to issues associated with global hunger, poverty, environmental sustainability, and resource consumption and provides…

  7. Real-time Texture Error Detection

    Directory of Open Access Journals (Sweden)

    Dan Laurentiu Lacrama

    2008-01-01

    Full Text Available This paper advocates an improved solution for the real-time error detection of texture errors that occurs in the production process in textile industry. The research is focused on the mono-color products with 3D texture model (Jacquard fabrics. This is a more difficult task than, for example, 2D multicolor textures.

  8. Real exchange rates in emerging economies

    Czech Academy of Sciences Publication Activity Database

    Brůha, J.; Podpiera, Jiří

    2010-01-01

    Roč. 18, č. 3 (2010), s. 599-628 ISSN 0967-0750 Institutional research plan: CEZ:AV0Z70850503 Keywords : real exchange rate * emerging economies * two-country model ling Subject RIV: AH - Economics Impact factor: 0.536, year: 2010

  9. 14 CFR 1273.31 - Real property.

    Science.gov (United States)

    2010-01-01

    ... percentage of participation in the cost of the original purchase to the fair market value of the property... GRANTS AND COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes... subgrantee's percentage of participation in the purchase of the real property to the current fair market...

  10. 21 CFR 1403.31 - Real property.

    Science.gov (United States)

    2010-04-01

    ... the cost of the original purchase to the fair market value of the property. However, in those... percentage of participation in the purchase of the real property to the current fair market value of the... COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards...

  11. 45 CFR 1174.31 - Real property.

    Science.gov (United States)

    2010-10-01

    ... the cost of the original purchase to the fair market value of the property. However, in those... percentage of participation in the purchase of the real property to the current fair market value of the... COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards...

  12. 45 CFR 1183.31 - Real property.

    Science.gov (United States)

    2010-10-01

    ... the cost of the original purchase to the fair market value of the property. However, in those... percentage of participation in the purchase of the real property to the current fair market value of the... COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards...

  13. 13 CFR 143.31 - Real property.

    Science.gov (United States)

    2010-01-01

    ... percentage of participation in the cost of the original purchase to the fair market value of the property... FOR GRANTS AND COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes... subgrantee's percentage of participation in the purchase of the real property to the current fair market...

  14. 45 CFR 1157.31 - Real property.

    Science.gov (United States)

    2010-10-01

    ... the cost of the original purchase to the fair market value of the property. However, in those... percentage of participation in the purchase of the real property to the current fair market value of the... AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes, Property, and Subawards § 1157.31...

  15. 28 CFR 66.31 - Real property.

    Science.gov (United States)

    2010-07-01

    ... percentage of participation in the cost of the original purchase to the fair market value of the property... AND COOPERATIVE AGREEMENTS TO STATE AND LOCAL GOVERNMENTS Post-Award Requirements Changes, Property... subgrantee's percentage of participation in the purchase of the real property to the current fair market...

  16. Numerical simulation of real-world flows

    Energy Technology Data Exchange (ETDEWEB)

    Hayase, Toshiyuki, E-mail: hayase@ifs.tohoku.ac.jp [Institute of Fluid Science, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577 (Japan)

    2015-10-15

    Obtaining real flow information is important in various fields, but is a difficult issue because measurement data are usually limited in time and space, and computational results usually do not represent the exact state of real flows. Problems inherent in the realization of numerical simulation of real-world flows include the difficulty in representing exact initial and boundary conditions and the difficulty in representing unstable flow characteristics. This article reviews studies dealing with these problems. First, an overview of basic flow measurement methodologies and measurement data interpolation/approximation techniques is presented. Then, studies on methods of integrating numerical simulation and measurement, namely, four-dimensional variational data assimilation (4D-Var), Kalman filters (KFs), state observers, etc are discussed. The first problem is properly solved by these integration methodologies. The second problem can be partially solved with 4D-Var in which only initial and boundary conditions are control parameters. If an appropriate control parameter capable of modifying the dynamical structure of the model is included in the formulation of 4D-Var, unstable modes are properly suppressed and the second problem is solved. The state observer and KFs also solve the second problem by modifying mathematical models to stabilize the unstable modes of the original dynamical system by applying feedback signals. These integration methodologies are now applied in simulation of real-world flows in a wide variety of research fields. Examples are presented for basic fluid dynamics and applications in meteorology, aerospace, medicine, etc. (topical review)

  17. Large Devaluations and the Real Exchange Rate

    Science.gov (United States)

    Burstein, Ariel; Eichenbaum, Martin; Rebelo, Sergio

    2005-01-01

    In this paper we argue that the primary force behind the large drop in real exchange rates that occurs after large devaluations is the slow adjustment in the prices of nontradable goods and services. Our empirical analysis uses data from five large devaluation episodes: Argentina (2002), Brazil (1999), Korea (1997), Mexico (1994), and Thailand…

  18. Teaching Real Business Cycles to Undergraduates

    Science.gov (United States)

    Brevik, Frode; Gartner, Manfred

    2007-01-01

    The authors review the graphical approach to teaching the real business cycle model introduced in Barro. They then look at where this approach cuts corners and suggest refinements. Finally, they compare graphical and exact models by means of impulse-response functions. The graphical models yield reliable qualitative results. Sizable quantitative…

  19. Real Business-cycle Model with Habits

    DEFF Research Database (Denmark)

    Khorunzhina, Natalia

    2015-01-01

    This paper empirically investigates the ability of a real business-cycle model with nonseparabilities in consumption and leisure and external habits both in consumption and leisure to fit the postwar US data. The results indicate a strong but fast-dying habit in leisure, and a somewhat weaker...

  20. Object recognition techniques in real applications

    NARCIS (Netherlands)

    Fernandez Robles, Laura

    2016-01-01

    This thesis proposes and evaluates object description and retrieval techniques in different real applications. First, we addressed the classification of boar spermatozoa according to acrosome integrity, which is an important challenge in the veterinary field. We presented several methods based on

  1. Real hypersurfaces of a complex projective space

    Indian Academy of Sciences (India)

    Kaehler manifolds of con- stant holomorphic sectional curvature c) is a very interesting and active area of research. The ambient space ¯M(c), specially in the case c = 0 imposes quite significant restrictions on the geometry of its real hypersurfaces.

  2. The homogeneous geometries of real hyperbolic space

    DEFF Research Database (Denmark)

    Castrillón López, Marco; Gadea, Pedro Martínez; Swann, Andrew Francis

    We describe the holonomy algebras of all canonical connections of homogeneous structures on real hyperbolic spaces in all dimensions. The structural results obtained then lead to a determination of the types, in the sense of Tricerri and Vanhecke, of the corresponding homogeneous tensors. We use ...

  3. Real Space Monitoring Technique in Ubiquitous Environments

    Science.gov (United States)

    Kon, Tetsuya; Ohira, Yutaka; Kubota, Yuzuru; Watanabe, Kazuhiro

    A method of a monitoring and remote controlling real world have been described by using a VRML/Java based virtual reality space which contains multimedia elements, intended for the development of a ubiquitous monitoring system. The focuses are given on the improvements for currently used, static virtual reality spaces to be more capable of an interactive function through which users can bilaterally monitor or control the real world. The monitoring is realized by reflecting sensing data from sensors which were placed in the real space onto a virtual reality space. Remote control can be realized by sending commands to actuators through the 3-D space. Additionally, the virtual reality space has been created as a Web page. So that the multiple users can access the system at the same time. The developed system has shown the availability for such applications as robot navigation, management of buildings and monitoring of factories and environments. In this paper, the construction method of the new technique of the real space monitoring and its operation experiment are described which can put in view a full-scall ubiquitous environment. The extendibility and possibility for monitoring are also described which could provide essential ubiquitous environments.

  4. Real Economies in Africa | Hardin | CODESRIA Bulletin

    African Journals Online (AJOL)

    Real Economies in Africa. Rebecca Hardin, Una Okonkwo. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL · AJOL's Partners · Terms and Conditions of Use · Contact AJOL · News. OTHER RESOURCES... for Researchers · for Journals · for Authors ...

  5. Competency Based Curriculum for Real Estate Agent.

    Science.gov (United States)

    McCloy, Robert J.

    This publication is a curriculum and teaching guide for preparing real estate agents in the state of West Virginia. The guide contains 30 units, or lessons. Each lesson is designed to cover three to five hours of instruction time. Competencies provided for each lesson are stated in terms of what the student should be able to do as a result of the…

  6. Cellphones and Real-World Communication

    Science.gov (United States)

    Bugeja, Michael

    2004-01-01

    In this article, the author shares his views on cellphones and real-world communication. He claims that the cellphone has changed society more than the home computer, which it has assimilated. Cellphones sound during worship, wakes, births, graduations, hearings, trials, and accreditation meetings--interrupting life-changing spiritual or secular…

  7. Three essays on real estate finance

    NARCIS (Netherlands)

    Liu, X.

    2010-01-01

    This thesis outlines several issues related to real estate research. The first chapter relaxes the functional form restriction in the empirical application of the hedonic pricing model, and studies its implication on the house price index construction. The second chapter takes into account both the

  8. Feedback as Real-Time Constructions

    Science.gov (United States)

    Keiding, Tina Bering; Qvortrup, Ane

    2014-01-01

    This article offers a re-description of feedback and the significance of time in feedback constructions based on systems theory. It describes feedback as internal, real-time constructions in a learning system. From this perspective, feedback is neither immediate nor delayed, but occurs in the very moment it takes place. This article argues for a…

  9. Real-Time Operating System/360

    Science.gov (United States)

    Hoffman, R. L.; Kopp, R. S.; Mueller, H. H.; Pollan, W. D.; Van Sant, B. W.; Weiler, P. W.

    1969-01-01

    RTOS has a cost savings advantage for real-time applications, such as those with random inputs requiring a flexible data routing facility, display systems simplified by a device independent interface language, and complex applications needing added storage protection and data queuing.

  10. Monetary Policy Frameworks and Real Equilibrium Determinacy

    DEFF Research Database (Denmark)

    Jensen, Henrik

    2002-01-01

    In a simple "prototype" model of monetary policymaking, I examine the issue of real equilibrium determinacy under targeting and instrument rules. The former framework involves minimization of a loss function (under discretion or commitment), whereas the latter involves commitment to an interest...

  11. Autopsies of the real: Resurrecting the dead

    Directory of Open Access Journals (Sweden)

    Valis, Noël

    2011-12-01

    Full Text Available The sense of the real, or the material—the dead body—as an inextricable part of the sacred does not disappear in the secular environment of the nineteenth and twentieth centuries. This article analyzes specific humanitarian narratives centered on the practice of autopsy and mummification, in which the traces of Catholicism act as a kind of spectral discourse of the imagination, where the real is configured in forms of the uncanny, the monstrous or the sacred.

    El sentido de lo real, de lo material —el cuerpo sin vida— como una inextricable parte de lo sagrado, no desaparece del ambiente secular de los siglos XIX y XX. En los relatos analizados en este artículo se estudia cómo en determinadas narrativas humanitarias centradas en la práctica de la autopsia y la momificación, las huellas del catolicismo actúan como una suerte de discurso espectral de la imaginación, en que lo real se configura en formas de lo siniestro, lo monstruoso o lo sagrado.

  12. Dentine hypersensitivity: real or imagined | Gbadebo | Nigerian ...

    African Journals Online (AJOL)

    However, the cause, diagnosis and possible management to give relief can be a dilemma for the clinician who at times may wonder if the sensation the individual is presenting with, is real or imagined. Aim: The purpose of this paper was to review dentine hypersensitivity in view of causes, diagnosis and management.

  13. What Is the Real Efficiency of Bulbs?

    Science.gov (United States)

    Polacek, Lubos

    2012-01-01

    Bulbs are considered to be very inefficient sources of light. Bulbs give light and heat. As we use them for a long time, especially in winter, a large part of the heat produced by bulbs lowers the power consumption of the heating system. In this paper the problem of the real efficiency of a bulb is solved when both the lighting and heating effects…

  14. Compiling models into real-time systems

    International Nuclear Information System (INIS)

    Dormoy, J.L.; Cherriaux, F.; Ancelin, J.

    1992-08-01

    This paper presents an architecture for building real-time systems from models, and model-compiling techniques. This has been applied for building a real-time model-base monitoring system for nuclear plants, called KSE, which is currently being used in two plants in France. We describe how we used various artificial intelligence techniques for building it: a model-based approach, a logical model of its operation, a declarative implementation of these models, and original knowledge-compiling techniques for automatically generating the real-time expert system from those models. Some of those techniques have just been borrowed from the literature, but we had to modify or invent other techniques which simply did not exist. We also discuss two important problems, which are often underestimated in the artificial intelligence literature: size, and errors. Our architecture, which could be used in other applications, combines the advantages of the model-based approach with the efficiency requirements of real-time applications, while in general model-based approaches present serious drawbacks on this point

  15. Snapple[R] Real Facts Watercolors

    Science.gov (United States)

    Hodge, Stephanie

    2007-01-01

    Did you know that a hummingbird weighs less than a penny? Or that an elephant swims twenty miles a day? These are just two of the many facts that Snapple prints on the lids of their drink bottles. When Snapple first introduced the Snapple Real Facts the author was excited to see that something so trivial was also educational. She asked herself if…

  16. Endangered Species: Real Life in Two Dimensions

    Science.gov (United States)

    Henderson, Lynette K.

    2012-01-01

    The focus of "Endangered Species: Real Life in Two Dimensions" is to create awareness about a critical environmental issue. There is a special urgency to this project because large numbers of animal species are currently endangered or on the brink of extinction. In addition to being enlightened about this important topic through research, students…

  17. Governance structures for real estate transactions: Markets ...

    African Journals Online (AJOL)

    This article examines the relative prevalence of markets, hierarchies and networks in the governance of real-estate transactions under three property rights regimes in Windhoek's low-income settlements. These governance structures are related to respective property rights regimes and to conjectures made about the ...

  18. Real Estate Curriculum for Community Colleges.

    Science.gov (United States)

    Nelson, Robert W.

    The Oregon Department of Education has prepared this curriculum guide to assist community college personnel in developing or upgrading real estate programs. This fast-growing field has demanded that community colleges analyze the course content of such programs so that they are relevant to the actual needs of the industry. An Advisory Committee…

  19. Real-time PCR gene expression profiling

    Czech Academy of Sciences Publication Activity Database

    Kubista, Mikael; Sjögreen, B.; Forootan, A.; Šindelka, Radek; Jonák, Jiří; Andrade, J.M.

    2007-01-01

    Roč. 1, - (2007), s. 56-60 ISSN 1360-8606 R&D Projects: GA AV ČR KJB500520601 Institutional research plan: CEZ:AV0Z50520514 Keywords : real - time PCR, * expression profiling * statistical analysis Subject RIV: EB - Genetics ; Molecular Biology

  20. Refactoring Real-Time Java Profiles

    DEFF Research Database (Denmark)

    Søndergaard, Hans; Thomsen, Bent; Ravn, Anders Peter

    2011-01-01

    Just like other software, Java profiles benefits from refactoring when they have been used and have evolved for some time. This paper presents a refactoring of the Real-Time Specification for Java (RTSJ) and the Safety Critical Java (SCJ) profile (JSR-302). It highlights core concepts and makes...... to comprehend and use for application developers and students....

  1. Robust synthesis for real-time systems

    DEFF Research Database (Denmark)

    Larsen, Kim Guldstrand; Legay, Axel; Traonouez, Louis-Marie

    2014-01-01

    Specification theories for real-time systems allow reasoning about interfaces and their implementation models, using a set of operators that includes satisfaction, refinement, logical and parallel composition. To make such theories applicable throughout the entire design process from an abstract...

  2. Real-time brute force SAR processing

    NARCIS (Netherlands)

    Vlothuizen, W.J.; Ditzel, M.

    2009-01-01

    This paper presents a brute force method to perform real-time SAR processing. The method has several advantages over traditional so-called fast SAR implementations, as it does not make any approximations to alleviate the processing burden. However, the method does allow efficient implementation on

  3. The real thing is something else

    DEFF Research Database (Denmark)

    Nielsen, Henrik Smed

    2008-01-01

    The trailer to Lady in the Lake (1947) promises an experience of the real which it will provide by having the camera assume Philip Marlowe's first person perspective throughout. Critics have discounted the experiment for a variety of reasons. Adopting a phenomenological perspective and comparing...

  4. Directing sustainable investments in commercial real estate

    NARCIS (Netherlands)

    Entrop, Alexis Gerardus; Brouwers, Jos; Braganca, L.; Pinheiro, M.; Jalali, S.; Mateus, R.; Amoeda, R.; Correia Guedes, M.

    2007-01-01

    This paper focuses on the facility costs, energy use and water consumption of commercial real estate. A framework consisting of four components is established in which the performances on these three aspects are analysed. By using the first, second and third part of the framework suggestions for

  5. Advances in Real-Time Systems

    CERN Document Server

    Chakraborty, Samarjit

    2012-01-01

    This volume contains the lectures given in honor to Georg Farber as tribute to his contributions in the area of real-time and embedded systems. The chapters of many leading scientists cover a wide range of aspects, like robot or automotive vision systems or medical aspects.

  6. Scene independent real-time indirect illumination

    DEFF Research Database (Denmark)

    Frisvad, Jeppe Revall; Christensen, Niels Jørgen; Falster, Peter

    2005-01-01

    A novel method for real-time simulation of indirect illumination is presented in this paper. The method, which we call Direct Radiance Mapping (DRM), is based on basal radiance calculations and does not impose any restrictions on scene geometry or dynamics. This makes the method tractable for rea...

  7. Real estate prices and bank stability

    NARCIS (Netherlands)

    Koetter, Michael; Poghosyan, Tigran

    Real estate prices can deviate from their fundamental value due to rigid supply, heterogeneity in quality, and various market imperfections, which have two contrasting effects on bank stability. Higher prices increase the value of collateral and net wealth of borrowers and thus reduce the likelihood

  8. Real Estate Appraising. A Suggested Adult Education Course.

    Science.gov (United States)

    New York State Education Dept., Albany. Bureau of Continuing Education Curriculum Development.

    Developed to assist teachers of classes in real estate appraisal and to help students prepare for professional qualification, this guide presents 12 lessons which may be taught in 15 two-hour sessions. The twelve lessons are (1) The Appraisal of Real Estate, (2) Real Property Value, (3) Basic Principles of Real Property Value, (4) The Appraisal…

  9. 12 CFR 614.4265 - Real property evaluations.

    Science.gov (United States)

    2010-01-01

    ... Collateral Evaluation Requirements § 614.4265 Real property evaluations. (a) Real estate shall be valued on... property and operation where the transaction value exceeds $250,000 and the real estate taken as collateral.... All other collateral associated with the real estate, but designated as personal property, shall be...

  10. Project Management in Real Time: A Service-Learning Project

    Science.gov (United States)

    Larson, Erik; Drexler, John A., Jr.

    2010-01-01

    This article describes a service-learning assignment for a project management course. It is designed to facilitate hands-on student learning of both the technical and the interpersonal aspects of project management, and it involves student engagement with real customers and real stakeholders in the creation of real events with real outcomes. As…

  11. Visualising the Complex Roots of Quadratic Equations with Real Coefficients

    Science.gov (United States)

    Bardell, Nicholas S.

    2012-01-01

    The roots of the general quadratic equation y = ax[superscript 2] + bx + c (real a, b, c) are known to occur in the following sets: (i) real and distinct; (ii) real and coincident; and (iii) a complex conjugate pair. Case (iii), which provides the focus for this investigation, can only occur when the values of the real coefficients a, b, and c are…

  12. 76 FR 78977 - Real Estate Settlement Procedures Act (Regulation X)

    Science.gov (United States)

    2011-12-20

    ... other assessments or charges on the real property; (14) Rendering of services by a real estate agent or... Part 1024 Real Estate Settlement Procedures Act (Regulation X); Interim Final Rule #0;#0;Federal... Development's (HUD's) rulemaking authority for the Real Estate Settlement Procedures Act (RESPA) to the Bureau...

  13. 7 CFR 1955.105 - Real property affected (CONACT).

    Science.gov (United States)

    2010-01-01

    ... inventory property. State Directors in consultation with other lenders, real estate agents, auctioneers, and... analyze farm real estate market conditions within the geographic areas of their jurisdiction and determine... real estate conditions. If the analyses of farm real estate conditions indicate that such sales would...

  14. 12 CFR 34.83 - Disposition of real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Disposition of real estate. 34.83 Section 34.83 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF THE TREASURY REAL ESTATE LENDING AND APPRAISALS Other Real Estate Owned § 34.83 Disposition of real estate. (a) Disposition. A national bank may...

  15. 12 CFR 541.16 - Improved residential real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Improved residential real estate. 541.16... REGULATIONS AFFECTING FEDERAL SAVINGS ASSOCIATIONS § 541.16 Improved residential real estate. The term improved residential real estate means residential real estate containing offsite or other improvements...

  16. 12 CFR 541.15 - Improved nonresidential real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Improved nonresidential real estate. 541.15... REGULATIONS AFFECTING FEDERAL SAVINGS ASSOCIATIONS § 541.15 Improved nonresidential real estate. The term improved nonresidential real estate means nonresidential real estate: (a) Containing a permanent structure...

  17. 33 CFR 211.22 - Real estate Claims Officers.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Real estate Claims Officers. 211..., DEPARTMENT OF DEFENSE REAL ESTATE ACTIVITIES OF THE CORPS OF ENGINEERS IN CONNECTION WITH CIVIL WORKS PROJECTS Real Estate Claims § 211.22 Real estate Claims Officers. In each Division Office the Division...

  18. 12 CFR 541.23 - Residential real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Residential real estate. 541.23 Section 541.23... AFFECTING FEDERAL SAVINGS ASSOCIATIONS § 541.23 Residential real estate. The terms residential real estate... home used in part for business); (c) Other real estate used for primarily residential purposes other...

  19. 32 CFR 644.30 - Preliminary real estate work.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Preliminary real estate work. 644.30 Section 644... PROPERTY REAL ESTATE HANDBOOK Project Planning Military (army and Air Force) and Other Federal Agencies § 644.30 Preliminary real estate work. (a) Preliminary real estate work is defined as that action taken...

  20. 7 CFR 1955.130 - Real estate brokers.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 14 2010-01-01 2009-01-01 true Real estate brokers. 1955.130 Section 1955.130... Dispose of Inventory Property § 1955.130 Real estate brokers. Contracting authority for the use of real... provide for any licensed real estate broker to provide sales services for any property listed under the...

  1. 12 CFR 34.62 - Real estate lending standards.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Real estate lending standards. 34.62 Section 34.62 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF THE TREASURY REAL ESTATE LENDING AND APPRAISALS Real Estate Lending Standards § 34.62 Real estate lending standards. (a) Each national bank shall...

  2. 12 CFR 365.2 - Real estate lending standards.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Real estate lending standards. 365.2 Section... POLICY REAL ESTATE LENDING STANDARDS § 365.2 Real estate lending standards. (a) Each insured state... extensions of credit that are secured by liens on or interests in real estate, or that are made for the...

  3. 12 CFR 541.27 - Unimproved real estate.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Unimproved real estate. 541.27 Section 541.27... AFFECTING FEDERAL SAVINGS ASSOCIATIONS § 541.27 Unimproved real estate. The term unimproved real estate means real estate that will be improved, as defined in § 541.15 or § 541.16 of this part. ...

  4. 33 CFR 211.2 - Authority to acquire real estate.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Authority to acquire real estate..., DEPARTMENT OF DEFENSE REAL ESTATE ACTIVITIES OF THE CORPS OF ENGINEERS IN CONNECTION WITH CIVIL WORKS PROJECTS Real Estate; General § 211.2 Authority to acquire real estate. (a) Congressional authority...

  5. Providing Real Research Opoportunities to Undergraduates

    Science.gov (United States)

    Ragozzine, Darin

    2016-01-01

    The current approach to undergraduate education focuses on teaching classes which provide the foundational knowledge for more applied experiences such as scientific research. Like most programs, Florida Institute of Technology (Florida Tech or FIT) strongly encourages undergraduate research, but is dominated by content-focused courses (e.g., "Physical Mechanics"). Research-like experiences are generally offered through "lab" classes, but these are almost always reproductions of past experiments: contrived, formulaic, and lacking the "heart" of real (i.e., potentially publishable) scientific research. Real research opportunities 1) provide students with realistic insight into the actual scientific process; 2) excite students far more than end-of-chapter problems; 3) provide context for the importance of learning math, physics, and astrophysics concepts; and 4) allow unique research progress for well-chosen problems. I have provided real research opportunities as an "Exoplanet Lab" component of my Introduction to Space Science (SPS1020) class at Florida Tech, generally taken by first-year majors in our Physics, Astronomy & Astrophysics, Planetary Science, and Astrobiology degree programs. These labs are a hybrid between citizen science (e.g., PlanetHunters) and simultaneously mentoring ~60 undergraduates in similar small research projects. These projects focus on problems that can be understood in the context of the course, but which benefit from "crowdsourcing". Examples include: dividing up the known planetary systems and developing a classification scheme and organizing them into populations (Fall 2013); searching through folded light curves to discover new exoplanets missed by previous pipelines (Fall 2014); and fitting n-body models to all exoplanets with known Transit Timing Variations to estimate planet masses (Fall 2015). The students love the fact that they are doing real potentially publishable research: not many undergraduates can claim to have discovered

  6. Argus developer in practice real estate development modeling in the real world

    CERN Document Server

    Havard, Tim M

    2014-01-01

    First ""missing manual"" for Argus Developer--case studies show readers how to analyze a development, something Argus manuals don't do. Argus Developer is by far the leading program for real estate developers worldwide The book is an education in real estate finance as well as the program Author teaches seminars and consults with people using the program--back of room sales likely Author has three other books on real estate development Possibility that Argus gets behind the project The book will contain dozens of screenshots

  7. Is there a Future in Real Estate? : Incorporate Futures Contracts within the Swiss Real Estate Market

    OpenAIRE

    Isberg, Ofelia

    2017-01-01

    In the past decades, real estate has turned into something more than just a home, it has become an investment. The interest to invest in the real estate market has increased from investors but also from private persons where the demand not only is to find a living but also perceived as an investment to make a profit or reinvest in the future.  The demand on the Swiss real estate market has increased from locals but also foreigners that want to invest in the stable and safe Swiss market. The h...

  8. Crowdsourced real-world sensing: sentiment analysis and the real-time web

    OpenAIRE

    Bermingham, Adam; Smeaton, Alan F.

    2010-01-01

    The advent of the real-time web is proving both challeng- ing and at the same time disruptive for a number of areas of research, notably information retrieval and web data mining. As an area of research reaching maturity, sentiment analysis oers a promising direction for modelling the text content available in real-time streams. This paper reviews the real-time web as a new area of focus for sentiment analysis and discusses the motivations and challenges behind such a direction.

  9. THE CONCEPT AND ESSENCE, THE STRUCTURE OF REAL ESTATE MARKET

    Directory of Open Access Journals (Sweden)

    Edgar V. Hovsepyan

    2015-01-01

    Full Text Available With the help of the international research literature, and the legislative framework of the Russian Federation and the Republic of Armenia in the article reveals the basic concepts and the essence of the real estate market, as well as given the classification of real estate. Thoroughly explains the difference between the concepts of "real estate" and "real property". In the article considered the main elements which includes the structure of the real estate market.

  10. Advanced real-time manipulation of video streams

    CERN Document Server

    Herling, Jan

    2014-01-01

    Diminished Reality is a new fascinating technology that removes real-world content from live video streams. This sensational live video manipulation actually removes real objects and generates a coherent video stream in real-time. Viewers cannot detect modified content. Existing approaches are restricted to moving objects and static or almost static cameras and do not allow real-time manipulation of video content. Jan Herling presents a new and innovative approach for real-time object removal with arbitrary camera movements.

  11. The touch of the real. Il New Historicism e il potere del reale psichiatrico in Foucault

    Directory of Open Access Journals (Sweden)

    VARRICCHIO, Francesco

    2014-12-01

    Full Text Available [The touch of the real. The New Historicism and the power of the psychiatric real in Foucault] What is the meaning of “touch of the real”? This paper connects the new historicist Greenblatt’s phrase to Foucault’s use of the term real during his lecture course on psychiatric power. According to New Historicism, influenced by the French philosopher’s thought, representation links to reality; but this reality is imbued with power relations. Throughout his course, Foucault repeteadly argues that therapeutic asylum imposes on subjectified bodiesthe reality of power and the power of reality thanks to a disciplinary technology. The constant visibility produces individuals also in other fields of the social and institutional life, in which the Panopticon principle is effective. The real springs from this mechanism and its representation.

  12. Real Estate Financing and Interest Rate Hedging : A quantitative real estate investment case study

    OpenAIRE

    van de Wiel, Wimjan; Kristopher Bock, Felix

    2017-01-01

    Background: The expansive monetary policy of the European Central Bank has been leading to all-time-low interest rates and to a strong move into real estate investment. Low interest rates can work in favor of the investor (due to low interest rate expenditures), but increasing interest rates can jeopardize real estate investments. Since changes in interest rates are unpredictable, an investor needs to deal with this volatility. The capital market offers several financial instruments (so-calle...

  13. Technical Qualifications for Treating Photovoltaic Assets as Real Property by Real Estate Investment Trusts (REITs)

    Energy Technology Data Exchange (ETDEWEB)

    Feldman, D.; Mendelsohn, M.; Coughlin, J.

    2012-06-01

    It has been proposed that Real Estate Investment Trusts (REITs) have the potential to lower the cost and increase the adoption of photovoltaic systems (PV) by offering a more attractive source of capital. The purpose of this paper is to explain the fundamental physical characteristics of PV and compare them to the characteristics of 'real' property, to help determine whether REITs can own PV systems.

  14. Acting to gain information: Real-time reasoning meets real-time perception

    Science.gov (United States)

    Rosenschein, Stan

    1994-01-01

    Recent advances in intelligent reactive systems suggest new approaches to the problem of deriving task-relevant information from perceptual systems in real time. The author will describe work in progress aimed at coupling intelligent control mechanisms to real-time perception systems, with special emphasis on frame rate visual measurement systems. A model for integrated reasoning and perception will be discussed, and recent progress in applying these ideas to problems of sensor utilization for efficient recognition and tracking will be described.

  15. Real time automatic discriminating of ultrasonic flaws

    International Nuclear Information System (INIS)

    Suhairy Sani; Mohd Hanif Md Saad; Marzuki Mustafa; Mohd Redzwan Rosli

    2009-01-01

    This paper is concerned with the real time automatic discriminating of flaws from two categories; i. cracks (planar defect) and ii. Non-cracks (volumetric defect such as cluster porosity and slag) using pulse-echo ultrasound. The raw ultrasonic flaws signal were collected from a computerized robotic plane scanning system over the whole of each reflector as the primary source of data. The signal is then filtered and the analysis in both time and frequency domain were executed to obtain the selected feature. The real time feature analysis techniques measured the number of peaks, maximum index, pulse duration, rise time and fall time. The obtained features could be used to distinguish between quantitatively classified flaws by using various tools in artificial intelligence such as neural networks. The proposed algorithm and complete system were implemented in a computer software developed using Microsoft Visual BASIC 6.0 (author)

  16. Applications of isotopy to real division algebras

    Energy Technology Data Exchange (ETDEWEB)

    Benkart, G. (Univ. of Wisconsin, Madison); Osborn, J.M.; Britten, D.

    1981-02-01

    In this paper we illustrate how the notion of isotopy can be used to solve various problems concerning finite-dimensional division algebras over the real numbers. In particular, we show that the 8-dimensional division algebras which have the same derivation algebra as the octonions, and hence which most resemble the octonions, are not in general isotopes of the octonions. Secondly, using a result of Hopf, we argue that every commutative division algebra is the reals or is isomorphic to a special kind of isotope of the complex numbers. Finally, by considering a certain class of algebras, we show how isotopy is a useful tool for determining necessary and sufficient conditions on the multiplication constants in order to have a division algebra.

  17. Real time gamma-ray signature identifier

    Science.gov (United States)

    Rowland, Mark [Alamo, CA; Gosnell, Tom B [Moraga, CA; Ham, Cheryl [Livermore, CA; Perkins, Dwight [Livermore, CA; Wong, James [Dublin, CA

    2012-05-15

    A real time gamma-ray signature/source identification method and system using principal components analysis (PCA) for transforming and substantially reducing one or more comprehensive spectral libraries of nuclear materials types and configurations into a corresponding concise representation/signature(s) representing and indexing each individual predetermined spectrum in principal component (PC) space, wherein an unknown gamma-ray signature may be compared against the representative signature to find a match or at least characterize the unknown signature from among all the entries in the library with a single regression or simple projection into the PC space, so as to substantially reduce processing time and computing resources and enable real-time characterization and/or identification.

  18. Analyzer of neutron flux in real time

    International Nuclear Information System (INIS)

    Rojas S, A.S.; Carrillo M, R.A.; Balderas, E.G.

    1999-01-01

    With base in the study of the real signals of neutron flux of instability events occurred in the Laguna Verde nuclear power plant where the nucleus oscillation phenomena of the reactor are in the 0 to 2.5 Hz range, it has been seen the possibility about the development a surveillance and diagnostic equipment capable to analyze in real time the behavior of nucleus in this frequencies range. An important method for surveillance the stability of the reactor nucleus is the use of the Power spectral density which allows to determine the frequencies and amplitudes contained in the signals. It is used an instrument carried out by LabVIEW graphic programming with a data acquisition card of 16 channels which works at Windows 95/98 environment. (Author)

  19. Real Space Approach to CMB deboosting

    CERN Document Server

    Yoho, Amanda; Starkman, Glenn D.; Pereira, Thiago S.

    2013-01-01

    The effect of our Galaxy's motion through the Cosmic Microwave Background rest frame, which aberrates and Doppler shifts incoming photons measured by current CMB experiments, has been shown to produce mode-mixing in the multipole space temperature coefficients. However, multipole space determinations are subject to many difficulties, and a real-space analysis can provide a straightforward alternative. In this work we describe a numerical method for removing Lorentz- boost effects from real-space temperature maps. We show that to deboost a map so that one can accurately extract the temperature power spectrum requires calculating the boost kernel at a finer pixelization than one might naively expect. In idealized cases that allow for easy comparison to analytic results, we have confirmed that there is indeed mode mixing among the spherical harmonic coefficients of the temperature. We find that using a boost kernel calculated at Nside=8192 leads to a 1% bias in the binned boosted power spectrum at l~2000, while ...

  20. RELATIONSHIP QUALITY IN REAL ESTATE COMPANIES

    Directory of Open Access Journals (Sweden)

    Florin Alexandru LUCA

    2016-05-01

    Full Text Available Customer loyalty is becoming a topic of interest which is winning increasingly more importance lately, as a result of the focus of marketing research on business relationships. One of the most important factors that determine customer loyalty is Service Quality.The stage of qualitative research consists of analyzing the research literature from this domain, which highlights the Relationship Quality construct as a distinct dimension, relevant for the real estate field. The summary of the main models of relationship quality indicate the most commonly used variables to describe this construct. The stage of quantitative research aims at testing the proportion of content validity for the most commonly used variables. The results describe an estimation of the dimensions of the Relationship Quality construct for the case of real estate services.

  1. Real symplectic formulation of local special geometry

    CERN Document Server

    Ferrara, Sergio; Ferrara, Sergio; Macia, Oscar

    2006-01-01

    We consider a formulation of local special geometry in terms of Darboux special coordinates $P^I=(p^i,q_i)$, $I=1,...,2n$. A general formula for the metric is obtained which is manifestly $\\mathbf{Sp}(2n,\\mathbb{R})$ covariant. Unlike the rigid case the metric is not given by the Hessian of the real function $S(P)$ which is the Legendre transform of the imaginary part of the holomorphic prepotential. Rather it is given by an expression that contains $S$, its Hessian and the conjugate momenta $S_I=\\frac{\\partial S}{\\partial P^I}$. Only in the one-dimensional case ($n=1$) is the real (two-dimensional) metric proportional to the Hessian with an appropriate conformal factor.

  2. Real-time sonography in obstetrics.

    Science.gov (United States)

    Anderson, S G

    1978-03-01

    Three hundred fifty real-time scans were performed on pregnant women for various indications. Placental localization was satisfactorily obtained in 173 of 174 studies. Estimates of fetal gestation from directly measured biparietal diameter were +/-2 weeks of actual gestation in 153 of 172 (88.9%) measurements. The presence or absence of fetal motion and cardiac activity established a diagnosis of fetal viability or fetal death in 32 patients after the first trimester. Accurate diagnosis was made in 52 of 57 patients with threatened abortions, and two of these errors occurred in scans performed before completion of the eighth postmenstrual week. Because of the ability to demonstrate fetal motion, real-time sonography should have many applications in obstetrics.

  3. Real Time Radiation Monitoring Using Nanotechnology

    Science.gov (United States)

    Li, Jing (Inventor); Wilkins, Richard T. (Inventor); Hanratty, James J. (Inventor); Lu, Yijiang (Inventor)

    2016-01-01

    System and method for monitoring receipt and estimating flux value, in real time, of incident radiation, using two or more nanostructures (NSs) and associated terminals to provide closed electrical paths and to measure one or more electrical property change values .DELTA.EPV, associated with irradiated NSs, during a sequence of irradiation time intervals. Effects of irradiation, without healing and with healing, of the NSs, are separately modeled for first order and second order healing. Change values.DELTA.EPV are related to flux, to cumulative dose received by NSs, and to radiation and healing effectivity parameters and/or.mu., associated with the NS material and to the flux. Flux and/or dose are estimated in real time, based on EPV change values, using measured .DELTA.EPV values. Threshold dose for specified changes of biological origin (usually undesired) can be estimated. Effects of time-dependent radiation flux are analyzed in pre-healing and healing regimes.

  4. Fobia: um sintoma marcado pelo real

    Directory of Open Access Journals (Sweden)

    Katja Linnemann

    Full Text Available O artigo pretende introduzir o questionamento acerca dos sintomas marcados pelo real, a partir da análise do sintoma fóbico do caso clínico de Freud, o “Pequeno Hans”. Por um lado, o sintoma como formação de compromisso (Freud, e como formação do inconsciente (Lacan, vem no lugar de algo que não pode ser sabido pelo sujeito, isto que é recalcado e endereçado ao Outro. Por outro, a fobia traz a particularidade de ser marcada por um real, carente de interdição paterna, sem visar um deciframento por parte do sujeito.

  5. Real estate space-territorial development

    Directory of Open Access Journals (Sweden)

    Sarchenko Vladimir Ivanovich

    2015-01-01

    Full Text Available Urbanization is one of the main sustainable tendencies of modern development. The contemporary state of real estate structure is characterized by high level of heterogeneity. In general, the consolidated design document of Moscow urban development, reconstruction and other forms of developing city territories doesn’t take into account the main tendencies of social and economic development, change of scales and structure of city production capacities, priorities of national economy development, or demand particularities, traditional and perspective preferences of citizens of different categories. The article focuses on systematization of basics of real estate space-territorial development strategic analysis. The author analyses the essence of analytical methods, their purpose, levels and analysis projections. The article emphasizes the estimation of economic effectiveness and revelation of urban environment development reserves.

  6. The real risk in pension forecasting

    DEFF Research Database (Denmark)

    Slipsager, Søren Kærgaard

    2018-01-01

    of the pension saver’s investment portfolio at retirement is highly dependent on the inflation scheme. As the deterministic scheme does not take state variable correlations into account it overestimates the expected portfolio value in real terms compared to the stochastic scheme. Moreover, the deterministic...... an alternative full model framework constructed around the three-factor diffusion model recently proposed by the Danish Society of Actuaries. By use of a simulation study the deterministic inflation scheme applied in the industry is compared to a stochastic scheme and it is shown that the real value...... scheme gives rise to a more heavy-tailed distribution implying a misestimation of downside risk and upside potential. Finally, it is shown in a realistic case study that the pension saver’s expected retirement payout profile is heavily affected....

  7. ENERGY CONSUMPTION AND REAL GDP IN IRAN

    Directory of Open Access Journals (Sweden)

    Ali Akbar Naji Meidani

    2014-01-01

    Full Text Available As one of the most important production factors and one of the most urgent final products, energy has a special position in the growth and development of the country. This paper examines the causal relationship between Real GDP and energy consumption in various economic sectors including (household and commercial, industry, transportation and agriculture sectors for Iran during 1967–2010 using the time series technique known as the Toda-Yamamoto method. Moreover, an error correction model is also estimated so that the results of these two methods are compared. We found a strong unidirectional causality from energy consumption in industry sector to real gross domestic product. Energy consumption in industry sector can observably promote the development of economy.

  8. The Fundamental Uncertainty of Business: Real Options

    Science.gov (United States)

    Dyer, James S.

    The purpose of this paper is to discuss the manner in which uncertainty is currently evaluated in business, with an emphasis on economic measures. In recent years, the accepted approach for the valuation of capital investment decisions has become one based on the theory of real options. From the standpoint of this workshop, the interesting aspect of real options is its focus on the flexibility of management to respond to changes in the environment as a feature of an alternative that has unique value, known as "option value." While this may not be surprising to most participants in this workshop, it does represent a radical change in traditional thinking about risk in business, where efforts have primarily been focused on the elimination of risk when possible.

  9. Tourism in Real and Virtual Space

    Directory of Open Access Journals (Sweden)

    Alireza Dehghan

    2009-07-01

    Full Text Available During these two decades, according to the expansion of communication, there is a deep transformation in individuals’ conception of space. As space plays an important role in tourism, either real or virtual, this transformation happens in the field too. The present study attempts to show how tourism in the contemporary virtualized world, or as some authors name: the dual globalized situation, occurs.

  10. ERRATARIUM for Real-Time Systems

    OpenAIRE

    Nelissen, Geoffrey

    2017-01-01

    Poster presented in 29th Euromicro Conference on Real-Time Systems (ECRTS 2017). 27 to 30, Jun, 2017, Interactive Session. Dubrovnik, Croatia. Hundreds of papers published each year • Some of them inevitably contain mistakes, errors, typos, … • All errors, big or small, deserve to be known by everyone • To avoid their spread in other works • To avoid teaching wrong results • To allow people to find appropriate solutions • To learn from them and not reproduce them later ...

  11. Real moments of the restrictive factor

    Indian Academy of Sciences (India)

    There are two cases here: 0 0 and a real number β such that. (1 + 2λ)/3 <β<(1 + 2λ)/3 + ϵ. In addition, we fix T ≤ x2, let α = 1 + c3(log x)−1. , and then deform the line of integration into the path which consists of the union of line segments. ⎧. ⎪⎪⎨.

  12. A real-time Global Warming Index

    OpenAIRE

    Haustein, K; Allen, MR; Forster, PM; Otto, FEL; Mitchell, DM; Matthews, HD; Frame, DJ

    2017-01-01

    We propose a simple real-time index of global human-induced warming and assess its robustness to uncertainties in climate forcing and short-term climate fluctuations. This index provides improved scientific context for temperature stabilisation targets and has the potential to decrease the volatility of climate policy. We quantify uncertainties arising from temperature observations, climate radiative forcings, internal variability and the model response. Our index and the associated rate of h...

  13. THE REAL OPTIONS OF CAPITAL BUDGET

    Directory of Open Access Journals (Sweden)

    Antonio Lopo Martins

    2008-07-01

    Full Text Available The traditional techniques of capital budget, as the deducted cash flow and the net value present, do not incorporate existing flexibilities in an investment project, they tend to distort the value of certain investments, mainly those that are considered in scenes of uncertainty and risk. Therefore, this study intends to demonstrate that the Real Options Theory (TOR is a useful methodology to evaluate and to indicate the best option for project of expansion investment. To reach the considered objective the procedure method was used a case study, having as unit of case the Resort Praia Hotel do Litoral Norte of Salvador. This study was developed of the following form: first it identified the traditional net value present and later it was incorporated the volatileness of each analyzed uncertainty. Second, as the real options are analogous to the financial options, it was necessary to identify elements that composed the terminologies of the financial options with intention to get the value of the real option. For this model of options pricing of Black & Scholes jointly with a computational simulator was used (SLS to get the expanded net value present. As a result of this study it was possible to evidence that using the traditional tool of capital budget Net Value Present (VPL is negative, therefore the project of expansion of the Hotel would be rejected. While for the application of methodology TOR the project presents positive Expanded Present Value which would represent an excellent chance of investment. Key-word: Capital budget, Real options, Analysis of investments.

  14. Real-time interactive treatment planning

    International Nuclear Information System (INIS)

    Otto, Karl

    2014-01-01

    The goal of this work is to develop an interactive treatment planning platform that permits real-time manipulation of dose distributions including DVHs and other dose metrics. The hypothesis underlying the approach proposed here is that the process of evaluating potential dose distribution options and deciding on the best clinical trade-offs may be separated from the derivation of the actual delivery parameters used for the patient’s treatment. For this purpose a novel algorithm for deriving an Achievable Dose Estimate (ADE) was developed. The ADE algorithm is computationally efficient so as to update dose distributions in effectively real-time while accurately incorporating the limits of what can be achieved in practice. The resulting system is a software environment for interactive real-time manipulation of dose that permits the clinician to rapidly develop a fully customized 3D dose distribution. Graphical navigation of dose distributions is achieved by a sophisticated method of identifying contributing fluence elements, modifying those elements and re-computing the entire dose distribution. 3D dose distributions are calculated in ∼2–20 ms. Including graphics processing overhead, clinicians may visually interact with the dose distribution (e.g. ‘drag’ a DVH) and display updates of the dose distribution at a rate of more than 20 times per second. Preliminary testing on various sites shows that interactive planning may be completed in ∼1–5 min, depending on the complexity of the case (number of targets and OARs). Final DVHs are derived through a separate plan optimization step using a conventional VMAT planning system and were shown to be achievable within 2% and 4% in high and low dose regions respectively. With real-time interactive planning trade-offs between Target(s) and OARs may be evaluated efficiently providing a better understanding of the dosimetric options available to each patient in static or adaptive RT. (paper)

  15. Real-time controller for hydrostatic transmission

    OpenAIRE

    2014-01-01

    M. Ing. (Electrical and Electronic Engineering) This dissertation describes the development of a modular real-time controller implemented on a personal computer for a hydrostatically driven vehicle. In such a vehicle the conventional mechanical transmission is replaced with a hydrostatic pump and two hydrostatic motors, making use of the secondary control principle. The infinitely variable transmission and wheel pair controller gives the vehicle superior traction and mobility over conventi...

  16. Real time closed orbit correction system

    International Nuclear Information System (INIS)

    Yu, L.H.; Biscardi, R.; Bittner, J.; Bozoki, E.; Galayda, J.; Krinsky, S.; Nawrocky, R.; Singh, O.; Vignola, G.

    1989-01-01

    We describe a global closed orbit feedback experiment, based upon a real time harmonic analysis of both the orbit movement and the correction magnetic fields. The feedback forces the coefficients of a few harmonics near the betatron tune to vanish, and significantly improves the global orbit stability. We present the results of the experiment in the UV ring using 4 detectors and 4 trims, in which maximum observed displacement was reduced by a factor of between 3 and 4. 4 refs., 3 figs

  17. Real Options Analysis of Mining Projects

    OpenAIRE

    Rudolf Zdravlje

    2011-01-01

    When long life assets are being evaluated based on constant predictions of future variables and the assumptions of zero management flexibility, is value being missed? In project evaluation today, the most common evaluation methods that calculate a net present value are discounted cash flow (DCF) analysis, decision tree analysis and Monte Carlo simulation. A fourth method, which is beginning to gain ground in terms of its use in the mining industry, is real option analysis (ROA). ROA utilizes ...

  18. Real-time optical information processing

    CERN Document Server

    Javidi, Bahram

    1994-01-01

    Real-Time Optical Information Processing covers the most recent developments in optical information processing, pattern recognition, neural computing, and materials for devices in optical computing. Intended for researchers and graduate students in signal and information processing with some elementary background in optics, the book provides both theoretical and practical information on the latest in information processing in all its aspects. Leading researchers in the field describe the significant signal processing algorithms architectures in optics as well as basic hardware concepts,

  19. Global Real Activity for Canadian Exports: GRACE

    OpenAIRE

    Binette, André; Chernis, Tony; De Munnik, Daniel

    2017-01-01

    Canadian exports have often disappointed since the Great Recession. The apparent disconnect between exports and the Bank of Canadafs current measure of foreign demand has created an impetus to search for an alternative. Based on a dynamic factor model (DFM) methodology, we use a broad range of international economic indicators (close to 300) to estimate external demand for Canadian exports. The new measure, Global Real Activity for Canadian Exports (GRACE), follows Binette et al. (201...

  20. Real closures of models of weak arithmetic

    Czech Academy of Sciences Publication Activity Database

    Jeřábek, Emil; Kolodziejczyk, L. A.

    2013-01-01

    Roč. 52, 1-2 (2013), s. 143-157 ISSN 0933-5846 R&D Projects: GA AV ČR IAA100190902; GA MŠk(CZ) 1M0545 Institutional support: RVO:67985840 Keywords : bounded arithmetic * real-closed field * recursive saturation Subject RIV: BA - General Mathematics http://link.springer.com/article/10.1007%2Fs00153-012-0311-x