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Sample records for single-copy real-time reverse-transcription

  1. 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.

  2. On-chip real-time single-copy polymerase chain reaction in picoliter droplets

    Energy Technology Data Exchange (ETDEWEB)

    Beer, N R; Hindson, B; Wheeler, E; Hall, S B; Rose, K A; Kennedy, I; Colston, B

    2007-04-20

    The first lab-on-chip system for picoliter droplet generation and PCR amplification with real-time fluorescence detection has performed PCR in isolated droplets at volumes 10{sup 6} smaller than commercial real-time PCR systems. The system utilized a shearing T-junction in a silicon device to generate a stream of monodisperse picoliter droplets that were isolated from the microfluidic channel walls and each other by the oil phase carrier. An off-chip valving system stopped the droplets on-chip, allowing them to be thermal cycled through the PCR protocol without droplet motion. With this system a 10-pL droplet, encapsulating less than one copy of viral genomic DNA through Poisson statistics, showed real-time PCR amplification curves with a cycle threshold of {approx}18, twenty cycles earlier than commercial instruments. This combination of the established real-time PCR assay with digital microfluidics is ideal for isolating single-copy nucleic acids in a complex environment.

  3. 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.

  4. Rapid and sensitive detection of canine distemper virus by real-time reverse transcription recombinase polymerase amplification.

    Science.gov (United States)

    Wang, Jianchang; Wang, Jinfeng; Li, Ruiwen; Liu, Libing; Yuan, Wanzhe

    2017-08-15

    Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic disease in free-living and captive carnivores worldwide. Recombinase polymerase amplification (RPA), as an isothermal gene amplification technique, has been explored for the molecular detection of diverse pathogens. A real-time reverse transcription RPA (RT-RPA) assay for the detection of canine distemper virus (CDV) using primers and exo probe targeting the CDV nucleocapsid protein gene was developed. A series of other viruses were tested by the RT-RPA.Thirty-two field samples were further tested by RT-RPA, and the resuts were compared with those obtained by the real-time RT-PCR. The RT-RPA assay was performed successfully at 40 °C, and the results were obtained within 3 min-12 min. The assay could detect CDV, but did not show cross-detection of canine parvovirus-2 (CPV-2), canine coronavirus (CCoV), canine parainfluenza virus (CPIV), pseudorabies virus (PRV) or Newcastle disease virus (NDV), demonstrating high specificity. The analytical sensitivity of RT-RPA was 31.8 copies in vitro transcribed CDV RNA, which is 10 times lower than the real-time RT-PCR. The assay performance was validated by testing 32 field samples and compared to real-time RT-PCR. The results indicated an excellent correlation between RT-RPA and a reference real-time RT-PCR method. Both assays provided the same results, and R 2 value of the positive results was 0.947. The results demonstrated that the RT-RPA assay offers an alternative tool for simple, rapid, and reliable detection of CDV both in the laboratory and point-of-care facility, especially in the resource-limited settings.

  5. Reference genes for gene expression analysis by real-time reverse transcription polymerase chain reaction of renal cell carcinoma.

    Science.gov (United States)

    Bjerregaard, Henriette; Pedersen, Shona; Kristensen, Søren Risom; Marcussen, Niels

    2011-12-01

    Differentiation between malignant renal cell carcinoma and benign oncocytoma is of great importance to choose the optimal treatment. Accurate preoperative diagnosis of renal tumor is therefore crucial; however, existing imaging techniques and histologic examinations are incapable of providing an optimal differentiation profile. Analysis of gene expression of molecular markers is a new possibility but relies on appropriate standardization to compare different samples. The aim of this study was to identify stably expressed reference genes suitable for the normalization of results extracted from gene expression analysis of renal tumors. Expression levels of 8 potential reference genes (ATP5J, HMBS, HPRT1, PPIA, TBP, 18S, GAPDH, and POLR2A) were examined by real-time reverse transcription polymerase chain reaction in tumor and normal tissue from removed kidneys from 13 patients with renal cell carcinoma and 5 patients with oncocytoma. The expression levels of genes were compared by gene stability value M, average gene stability M, pairwise variation V, and coefficient of variation CV. More candidates were not suitable for the purpose, but a combination of HMBS, PPIA, ATP5J, and TBP was found to be the best combination with an average gene stability value M of 0.9 and a CV of 0.4 in the 18 tumors and normal tissues. A combination of 4 genes, HMBS, PPIA, ATP5J, and TBP, is a possible reference in renal tumor gene expression analysis by reverse transcription polymerase chain reaction. A combination of four genes, HMBS, PPIA, ATP5J and TBP, being stably expressed in tissues from RCC is possible reference genes for gene expression analysis.

  6. Evaluation of a duplex reverse-transcription real-time PCR assay for the detection of encephalomyocarditis virus.

    Science.gov (United States)

    Qin, Shaomin; Underwood, Darren; Driver, Luke; Kistler, Carol; Diallo, Ibrahim; Kirkland, Peter D

    2018-06-01

    We evaluated a fluorogenic probe-based assay for the detection of encephalomyocarditis virus (EMCV) by comparing a set of published primers and probe to a new set of primers and probe. The published reagents failed to amplify a range of Australian isolates and an Italian reference strain of EMCV. In contrast, an assay based on 2 new sets of primers and probes that were run in a duplex reverse-transcription real-time PCR (RT-rtPCR) worked well, with high amplification efficiency. The analytical sensitivity was ~100-fold higher than virus isolation in cell culture. The intra-assay variation was 0.21-4.90%. No cross-reactivity was observed with a range of other porcine viruses. One hundred and twenty-two clinical specimens were tested simultaneously by RT-rtPCR and virus isolation in cell culture; 72 specimens gave positive results by RT-rtPCR, and 63 of these were also positive by virus isolation. Of 245 archived cell culture isolates of EMCV that were tested in the RT-rtPCR, 242 samples were positive. The new duplex RT-rtPCR assay is a reliable tool for the detection of EMCV in clinical specimens and for use in epidemiologic investigations.

  7. Exogenous reference gene normalization for real-time reverse transcription-polymerase chain reaction analysis under dynamic endogenous transcription.

    Science.gov (United States)

    Johnston, Stephen; Gallaher, Zachary; Czaja, Krzysztof

    2012-05-15

    Quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is widely used to investigate transcriptional changes following experimental manipulations to the nervous system. Despite the widespread utilization of qPCR, the interpretation of results is marred by the lack of a suitable reference gene due to the dynamic nature of endogenous transcription. To address this inherent deficiency, we investigated the use of an exogenous spike-in mRNA, luciferase, as an internal reference gene for the 2(-∆∆Ct) normalization method. To induce dynamic transcription, we systemically administered capsaicin, a neurotoxin selective for C-type sensory neurons expressing the TRPV-1 receptor, to adult male Sprague-Dawley rats. We later isolated nodose ganglia for qPCR analysis with the reference being either exogenous luciferase mRNA or the commonly used endogenous reference β-III tubulin. The exogenous luciferase mRNA reference clearly demonstrated the dynamic expression of the endogenous reference. Furthermore, variability of the endogenous reference would lead to misinterpretation of other genes of interest. In conclusion, traditional reference genes are often unstable under physiologically normal situations, and certainly unstable following the damage to the nervous system. The use of exogenous spike-in reference provides a consistent and easily implemented alternative for the analysis of qPCR data.

  8. Outbreak of hepatitis E virus infection in Darfur, Sudan: effectiveness of real-time reverse transcription-PCR analysis of dried blood spots.

    Science.gov (United States)

    Mérens, Audrey; Guérin, Philippe Jean; Guthmann, Jean-Paul; Nicand, Elisabeth

    2009-06-01

    Biological samples collected in refugee camps during an outbreak of hepatitis E were used to compare the accuracy of hepatitis E virus RNA amplification by real-time reverse transcription-PCR (RT-PCR) for sera and dried blood spots (concordance of 90.6%). Biological profiles (RT-PCR and serology) of asymptomatic individuals were also analyzed.

  9. Real-time reverse transcription-polymerase chain reaction assays for identification of wild poliovirus 1 & 3.

    Science.gov (United States)

    Sharma, Deepa K; Nalavade, Uma P; Deshpande, Jagadish M

    2015-10-01

    The poliovirus serotype identification and intratypic differentiation by real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay is suitable for serotype mixtures but not for intratypic mixtures of wild and vaccine poliovirus strains. This study was undertaken to develop wild poliovirus 1 and 3 (WPV1 and WPV3) specific rRT-PCR assays for use. Specific primers and probes for rRT-PCR were designed based on VP1 sequences of WPV1 and WPV3 isolated in India since 2000. The specificity of the rRT-PCR assays was evaluated using WPV1 and WPV3 of different genetic lineages, non-polio enteroviruses (NPEVs) and mixtures of wild/wild and wild/Sabin vaccine strains. The sensitivity of the assays was determined by testing serial 10-fold dilutions of wild poliovirus 1 and 3 stock suspensions of known titre. No cross-reactivity with Sabin strains, intertypic wild poliovirus isolates or 27 types of NPEVs across all the four Enterovirus species was found for both the wild poliovirus 1 and 3 rRT-PCR assays. All WPV1 and WPV3 strains isolated since 2000 were successfully amplified. The rRT-PCR assays detected 10 4.40 CCID 50 /ml of WPV1 and 10 4.00 CCID 50 /ml of WPV3, respectively either as single isolate or mixture with Sabin vaccine strains or intertypic wild poliovirus. rRT-PCR assays for WPV1 and WPV3 have been validated to detect all the genetic variations of the WPV1 and WPV3 isolated in India for the last decade. When used in combination with the current rRT-PCR assay testing was complete for confirmation of the presence of wild poliovirus in intratypic mixtures.

  10. Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection.

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    Laurent Dacheux

    2016-07-01

    Full Text Available The definitive diagnosis of lyssavirus infection (including rabies in animals and humans is based on laboratory confirmation. The reference techniques for post-mortem rabies diagnosis are still based on direct immunofluorescence and virus isolation, but molecular techniques, such as polymerase chain reaction (PCR based methods, are increasingly being used and now constitute the principal tools for diagnosing rabies in humans and for epidemiological analyses. However, it remains a key challenge to obtain relevant specificity and sensitivity with these techniques while ensuring that the genetic diversity of lyssaviruses does not compromise detection. We developed a dual combined real-time reverse transcription polymerase chain reaction (combo RT-qPCR method for pan-lyssavirus detection. This method is based on two complementary technologies: a probe-based (TaqMan RT-qPCR for detecting the RABV species (pan-RABV RT-qPCR and a second reaction using an intercalating dye (SYBR Green to detect other lyssavirus species (pan-lyssa RT-qPCR. The performance parameters of this combined assay were evaluated with a large panel of primary animal samples covering almost all the genetic variability encountered at the viral species level, and they extended to almost all lyssavirus species characterized to date. This method was also evaluated for the diagnosis of human rabies on 211 biological samples (positive n = 76 and negative n = 135 including saliva, skin and brain biopsies. It detected all 41 human cases of rabies tested and confirmed the sensitivity and the interest of skin biopsy (91.5% and saliva (54% samples for intra-vitam diagnosis of human rabies. Finally, this method was successfully implemented in two rabies reference laboratories in enzootic countries (Cambodia and Morocco. This combined RT-qPCR method constitutes a relevant, useful, validated tool for the diagnosis of rabies in both humans and animals, and represents a promising tool for

  11. Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection.

    Science.gov (United States)

    Dacheux, Laurent; Larrous, Florence; Lavenir, Rachel; Lepelletier, Anthony; Faouzi, Abdellah; Troupin, Cécile; Nourlil, Jalal; Buchy, Philippe; Bourhy, Herve

    2016-07-01

    The definitive diagnosis of lyssavirus infection (including rabies) in animals and humans is based on laboratory confirmation. The reference techniques for post-mortem rabies diagnosis are still based on direct immunofluorescence and virus isolation, but molecular techniques, such as polymerase chain reaction (PCR) based methods, are increasingly being used and now constitute the principal tools for diagnosing rabies in humans and for epidemiological analyses. However, it remains a key challenge to obtain relevant specificity and sensitivity with these techniques while ensuring that the genetic diversity of lyssaviruses does not compromise detection. We developed a dual combined real-time reverse transcription polymerase chain reaction (combo RT-qPCR) method for pan-lyssavirus detection. This method is based on two complementary technologies: a probe-based (TaqMan) RT-qPCR for detecting the RABV species (pan-RABV RT-qPCR) and a second reaction using an intercalating dye (SYBR Green) to detect other lyssavirus species (pan-lyssa RT-qPCR). The performance parameters of this combined assay were evaluated with a large panel of primary animal samples covering almost all the genetic variability encountered at the viral species level, and they extended to almost all lyssavirus species characterized to date. This method was also evaluated for the diagnosis of human rabies on 211 biological samples (positive n = 76 and negative n = 135) including saliva, skin and brain biopsies. It detected all 41 human cases of rabies tested and confirmed the sensitivity and the interest of skin biopsy (91.5%) and saliva (54%) samples for intra-vitam diagnosis of human rabies. Finally, this method was successfully implemented in two rabies reference laboratories in enzootic countries (Cambodia and Morocco). This combined RT-qPCR method constitutes a relevant, useful, validated tool for the diagnosis of rabies in both humans and animals, and represents a promising tool for lyssavirus

  12. Screening suitable reference genes for normalization in reverse transcription quantitative real-time PCR analysis in melon.

    Directory of Open Access Journals (Sweden)

    Qiusheng Kong

    Full Text Available Melon (Cucumis melo. L is not only an economically important cucurbitaceous crop but also an attractive model for studying many biological characteristics. Screening appropriate reference genes is essential to reverse transcription quantitative real-time PCR (RT-qPCR, which is key to many studies involving gene expression analysis. In this study, 14 candidate reference genes were selected, and the variations in their expression in roots and leaves of plants subjected to biotic stress, abiotic stress, and plant growth regulator treatment were assessed by RT-qPCR. The stability of the expression of the selected genes was determined and ranked using geNorm and NormFinder. geNorm identified the two most stable genes for each set of conditions: CmADP and CmUBIep across all samples, CmUBIep and CmRPL in roots, CmRAN and CmACT in leaves, CmADP and CmRPL under abiotic stress conditions, CmTUA and CmACT under biotic stress conditions, and CmRAN and CmACT under plant growth regulator treatments. NormFinder determined CmRPL to be the best reference gene in roots and under biotic stress conditions and CmADP under the other experimental conditions. CmUBC2 and CmPP2A were not found to be suitable under many experimental conditions. The catalase family genes CmCAT1, CmCAT2, and CmCAT3 were identified in melon genome and used as target genes to validate the reliability of identified reference genes. The catalase family genes showed the most upregulation 3 days after inoculation with Fusarium wilt in roots, after which they were downregulated. Their levels of expression were significantly overestimated when the unsuitable reference gene was used for normalization. These results not only provide guidelines for the selection of reference genes for gene expression analyses in melons but may also provide valuable information for studying the functions of catalase family genes in stress responses.

  13. Enteroviruses in blood of patients with type 1 diabetes detected by integrated cell culture and reverse transcription quantitative real-time PCR.

    Science.gov (United States)

    Alidjinou, Enagnon Kazali; Sane, Famara; Lefevre, Christine; Baras, Agathe; Moumna, Ilham; Engelmann, Ilka; Vantyghem, Marie-Christine; Hober, Didier

    2017-11-01

    Enteroviruses (EV) have been associated with type 1 diabetes (T1D), but EV RNA detection has been reported in only a small proportion of T1D patients. We studied whether integrated cell culture and reverse transcription real-time PCR could improve EV detection in blood samples from patients with T1D. Blood was collected from 13 patients with T1D. The presence of EV RNA in blood was investigated by using real-time RT-PCR. In addition, plasma and white blood cells (WBC) were inoculated to BGM and Vero cell line cultures. Culture supernatants and cells collected on day 7 and day 14 were tested for EV RNA by real-time RT-PCR. Enterovirus identification was performed through sequencing of the VP4/VP2 region. Enterovirus RNA was detected in blood by using real-time RT-PCR in only one out of 13 patients. The detection of EV RNA in cultures inoculated with clinical samples (plasma and/or WBC) gave positive results in five other patients. The viral loads were low, ranging from 45 to 4420 copies/ng of total RNA. One isolate was successfully identified as coxsackievirus B1. Integrated cell culture and reverse transcription real-time PCR can improve the detection rate of EV in blood samples of patients with T1D and can be useful to investigate further the relationship between EV and the disease.

  14. Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Son, Na Ry; Seo, Dong Joo; Lee, Min Hwa; Seo, Sheungwoo; Wang, Xiaoyu; Lee, Bog-Hieu; Lee, Jeong-Su; Joo, In-Sun; Hwang, In-Gyun; Choi, Changsun

    2014-09-01

    The aim of this study was to develop an optimal technique for detecting hepatitis E virus (HEV) in swine livers. Here, three elution buffers and two concentration methods were compared with respect to enhancing recovery of HEV from swine liver samples. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested RT-PCR were performed to detect HEV RNA. When phosphate-buffered saline (PBS, pH 7.4) was used to concentrate HEV in swine liver samples using ultrafiltration, real-time RT-PCR detected HEV in 6 of the 26 samples. When threonine buffer was used to concentrate HEV using polyethylene glycol (PEG) precipitation and ultrafiltration, real-time RT-PCR detected HEV in 1 and 3 of the 26 samples, respectively. When glycine buffer was used to concentrate HEV using ultrafiltration and PEG precipitation, real-time RT-PCR detected HEV in 1 and 3 samples of the 26 samples, respectively. When nested RT-PCR was used to detect HEV, all samples tested negative regardless of the type of elution buffer or concentration method used. Therefore, the combination of real-time RT-PCR and ultrafiltration with PBS buffer was the most sensitive and reliable method for detecting HEV in swine livers. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Porcine reproductive and respiratory syndrome virus: Interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods

    DEFF Research Database (Denmark)

    Wernike, Kerstin; Bonilauri, Paolo; Dauber, Malte

    2012-01-01

    To compare the real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays used for the diagnosis of Porcine reproductive and respiratory syndrome virus (PRRSV), a Europe-wide interlaboratory ring trial was conducted. A variety of PRRSV strains including North American...... (NA) and European (EU) genotype isolates were analyzed by the participants. Great differences regarding qualitative diagnostics as well as analytical sensitivity were observed between the individual RT-qPCR systems, especially when investigating strains from the EU genotype. None of the assays...

  16. Development of field-based real-time reverse transcription-polymerase chain reaction assays for detection of Chikungunya and O'nyong-nyong viruses in mosquitoes.

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    Smith, Darci R; Lee, John S; Jahrling, Jordan; Kulesh, David A; Turell, Michael J; Groebner, Jennifer L; O'Guinn, Monica L

    2009-10-01

    Chikungunya (CHIK) and O'nyong-nyong (ONN) are important emerging arthropod-borne diseases. Molecular diagnosis of these two viruses in mosquitoes has not been evaluated, and the effects of extraneous mosquito tissue on assay performance have not been tested. Additionally, no real-time reverse transcription-polymerase chain reaction (RT-PCR) assay exists for detecting ONN virus (ONNV) RNA. We describe the development of sensitive and specific real-time RT-PCR assays for detecting CHIK and ONN viral RNA in mosquitoes, which have application for field use. In addition, we compared three methods for primer/probe design for assay development by evaluating their sensitivity and specificity. This comparison resulted in development of virus-specific assays that could detect less than one plaque-forming unit equivalent of each of the viruses in mosquitoes. The use of these assays will aid in arthropod-borne disease surveillance and in the control of the associated diseases.

  17. Variation in Bluetongue virus real-time reverse transcription polymerase chain reaction assay results in blood samples of sheep, cattle, and alpaca.

    Science.gov (United States)

    Brito, Barbara P; Gardner, Ian A; Hietala, Sharon K; Crossley, Beate M

    2011-07-01

    Bluetongue is a vector-borne viral disease that affects domestic and wild ruminants. The epidemiology of this disease has recently changed, with occurrence in new geographic areas. Various real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) assays are used to detect Bluetongue virus (BTV); however, the impact of biologic differences between New World camelids and domestic ruminant samples on PCR efficiency, for which the BTV real-time qRT-PCR was initially validated are unknown. New world camelids are known to have important biologic differences in whole blood composition, including hemoglobin concentration, which can alter PCR performance. In the present study, sheep, cattle, and alpaca blood were spiked with BTV serotypes 10, 11, 13, and 17 and analyzed in 10-fold dilutions by real-time qRT-PCR to determine if species affected nucleic acid recovery and assay performance. A separate experiment was performed using spiked alpaca blood subsequently diluted in 10-fold series in sheep blood to assess the influence of alpaca blood on performance efficiency of the BTV real-time qRT-PCR assay. Results showed that BTV-specific nucleic acid detection from alpaca blood was consistently 1-2 logs lower than from sheep and cattle blood, and results were similar for each of the 4 BTV serotypes analyzed.

  18. Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results.

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    Andreas Garcia-Bardon

    Full Text Available Real-time reverse transcription polymerase chain reaction (PCR is the gold standard for expression analysis. Designed to improve reproducibility and sensitivity, commercial kits are commonly used for the critical step of cDNA synthesis. The present study was designed to determine the impact of these kits. mRNA from mouse brains were pooled to create serial dilutions ranging from 0.0625 μg to 2 μg, which were transcribed into cDNA using four different commercial reverse-transcription kits. Next, we transcribed mRNA from brain tissue after acute brain injury and naïve mice into cDNA for qPCR. Depending on tested genes, some kits failed to show linear results in dilution series and revealed strong variations in cDNA yield. Absolute expression data in naïve and trauma settings varied substantially between these kits. Normalization with a housekeeping gene failed to reduce kit-dependent variations, whereas normalization eliminated differences when naïve samples from the same region were used. The study shows strong evidence that choice of commercial cDNA synthesis kit has a major impact on PCR results and, consequently, on comparability between studies. Additionally, it provides a solution to overcome this limitation by normalization with data from naïve samples. This simple step helps to compare mRNA expression data between different studies and groups.

  19. Laboratory evaluation of a quantitative real-time reverse transcription PCR assay for the detection and identification of the four subgroups of avian metapneumovirus.

    Science.gov (United States)

    Guionie, O; Toquin, D; Sellal, E; Bouley, S; Zwingelstein, F; Allée, C; Bougeard, S; Lemière, S; Eterradossi, N

    2007-02-01

    Avian metapneumovirus (AMPV) is an important pathogen causing respiratory diseases and egg drops in several avian species. Four AMPV subgroups have been identified. The laboratory diagnosis of AMPV infections relies on serological methods, on labour-intensive virus isolation procedures, and on recently developed subgroup specific reverse transcription PCR (RT-PCR) protocols. In the present study, both the specificity and sensitivity of a commercial real-time reverse transcription PCR (RRT-PCR) for the detection and identification of the four AMPV subgroups were evaluated. Fifteen non-AMPV avian viruses belonging to 7 genera and 32 AMPV belonging to the 4 subgroups were tested. No non-AMPV virus was detected, whereas all AMPV viruses were identified in agreement with their previous molecular and antigenic subgroup assignment. The sensitivity and quantitating ability of the RRT-PCR assay were determined using serial dilutions of RNA derived either from AMPV virus stocks or from runoff transcripts. In all cases, linear dose/responses were observed. The detection limits of the different subgroups ranged from 500 to 5000 RNA copies and from 0.03 to 3.16TCID50/ml. The results were reproducible under laboratory conditions, thus showing that quantitative RRT-PCR is a new and powerful tool for the rapid and sensitive detection, identification and quantitation of AMPVs.

  20. Improved Safety for Molecular Diagnosis of Classical Rabies Viruses by Use of a TaqMan Real-Time Reverse Transcription-PCR "Double Check" Strategy

    DEFF Research Database (Denmark)

    Hoffmann, B.; Freuling, C. M.; Wakeley, P. R.

    2010-01-01

    To improve the diagnosis of classical rabies virus with molecular methods, a validated, ready-to-use, real-time reverse transcription-PCR (RT-PCR) assay was developed. In a first step, primers and 6-carboxyfluorescien-labeled TaqMan probes specific for rabies virus were selected from the consensus...... sequence of the nucleoprotein gene of 203 different rabies virus sequences derived from GenBank. The selected primer-probe combination was highly specific and sensitive. During validation using a sample set of rabies virus strains from the virus archives of the Friedrich-Loeffler-Institut (FLI; Germany......), the Veterinary Laboratories Agency (VLA; United Kingdom), and the DTU National Veterinary Institute (Lindholm, Denmark), covering the global diversity of rabies virus lineages, it was shown that both the newly developed assay and a previously described one had some detection failures. This was overcome...

  1. Application of a Real-time Reverse Transcription Loop Mediated Amplification Method to the Detection of Rabies Virus in Arctic Foxes in Greenland

    DEFF Research Database (Denmark)

    Wakeley, Philip; Johnson, Nicholas; Rasmussen, Thomas Bruun

    Reverse transcription loop mediated amplification (RT-LAMP) offers a rapid, isothermal method for amplification of virus RNA. In this study a panel of positive rabies virus samples originally prepared from arctic fox brain tissue was assessed for the presence of rabies viral RNA using a real time...... RT-LAMP. The method had previously been shown to work with samples from Ghana which clustered with cosmopolitan lineage rabies viruses but the assay had not been assessed using samples from animals infected with rabies from the arctic region. The assay is designed to amplify both cosmopolitan strains...... and arctic-like strains of classical rabies virus due to the primer design and is therefore expected to be universally applicable independent of region of the world where the virus is isolated. Of the samples tested all were found to be positive after incubation for 25 to 30 minutes. The method made use...

  2. Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains.

    Science.gov (United States)

    Wilkes, Rebecca P; Sanchez, Elena; Riley, Matthew C; Kennedy, Melissa A

    2014-01-01

    Canine distemper virus (CDV) remains a common cause of infectious disease in dogs, particularly in high-density housing situations such as shelters. Vaccination of all dogs against CDV is recommended at the time of admission to animal shelters and many use a modified live virus (MLV) vaccine. From a diagnostic standpoint for dogs with suspected CDV infection, this is problematic because highly sensitive diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) tests are able to detect MLV virus in clinical samples. Real-time PCR can be used to quantitate amount of virus shedding and can differentiate vaccine strains from wild-type strains when shedding is high. However, differentiation by quantitation is not possible in vaccinated animals during acute infection, when shedding is low and could be mistaken for low level vaccine virus shedding. While there are gel-based RT-PCR assays for differentiation of vaccine strains from field strains based on sequence differences, the sensitivity of these assays is unable to match that of the real-time RT-PCR assay currently used in the authors' laboratory. Therefore, a real-time RT-PCR assay was developed that detects CDV MLV vaccine strains and distinguishes them from wild-type strains based on nucleotide sequence differences, rather than the amount of viral RNA in the sample. The test is highly sensitive, with detection of as few as 5 virus genomic copies (corresponding to 10(-1) TCID(50)). Sequencing of the DNA real-time products also allows phylogenetic differentiation of the wild-type strains. This test will aid diagnosis during outbreaks of CDV in recently vaccinated animals.

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

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    Marilia Farignoli Romeiro

    2016-06-01

    Full Text Available Abstract: INTRODUCTION: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

  4. Comparison of the Diagnostic Value Between Real-Time Reverse Transcription-Polymerase Chain Reaction Assay and Histopathologic Examination in Sentinel Lymph Nodes for Patients With Gastric Carcinoma.

    Science.gov (United States)

    Kwak, Yoonjin; Nam, Soo Kyung; Shin, Eun; Ahn, Sang-Hoon; Lee, Hee Eun; Park, Do Joong; Kim, Woo Ho; Kim, Hyung-Ho; Lee, Hye Seung

    2016-05-01

    Sentinel lymph node (SLN)-based diagnosis in gastric cancers has shown varied sensitivities and false-negative rates in several studies. Application of the reverse transcription-polymerase chain reaction (RT-PCR) in SLN diagnosis has recently been proposed. A total of 155 SLNs from 65 patients with cT1-2, N0 gastric cancer were examined. The histopathologic results were compared with results obtained by real-time RT-PCR for detecting molecular RNA (mRNA) of cytokeratin (CK)19, carcinoembryonic antigen (CEA), and CK20. The sensitivity and specificity of the multiple marker RT-PCR assay standardized against the results of the postoperative histological examination were 0.778 (95% confidence interval [CI], 0.577-0.914) and 0.781 (95% CI, 0.700-0.850), respectively. In comparison, the sensitivity and specificity of intraoperative diagnosis were 0.819 (95% CI, 0.619-0.937) and 1.000 (95% CI, 0.972-1.000), respectively. The positive predictive value of the multiple-marker RT-PCR assay was 0.355 (95% CI, 0.192-0.546) for predicting non-SLN metastasis, which was lower than that of intraoperative diagnosis (0.813, 95% CI, 0.544-0.960). The real-time RT-PCR assay could detect SLN metastasis in gastric cancer. However, the predictive value of the real-time RT-PCR assay was lower than that of precise histopathologic examination and did not outweigh that of our intraoperative SLN diagnosis. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. Comparison of reverse-transcription real-time PCR and immunohistochemistry for the detection of canine distemper virus infection in raccoons in Ontario, Canada.

    Science.gov (United States)

    Nemeth, Nicole M; Oesterle, Paul T; Campbell, G Douglas; Ojkic, Davor; Jardine, Claire M

    2018-03-01

    Canine distemper virus (CDV) is a widespread morbillivirus that causes subclinical to fatal infections in domestic and wild carnivores. Raccoons ( Procyon lotor) are CDV reservoirs and suffer from associated disease. Aspects of pathogenesis may lead to difficulty in the interpretation of commonly used testing modalities, such as reverse-transcription real-time (RT-rt)PCR and immunohistochemistry (IHC). The reliance upon such tests is greater for wildlife, which are often submitted as carcasses with no clinical history. We compared CDV RT-rtPCR results to immunohistochemistry (the gold standard) in tissues from 74 raccoons. These tests had high kappa agreement (lymph node: 0.9335; lung: 0.8671) and a negative correlation between IHC score and threshold cycle (Ct) value for lymph node and lung (Spearman rank correlation coefficient [ r s ] = -0.8555 and -0.8179, respectively; p < 0.00001). An RT-rtPCR Ct value of 30 in lung and lymph node with sensitivity and specificity of 92.3 and 92.6% and 86.8 and 96.4%, respectively, was suitable for determining CDV involvement. Conjunctival swabs provide an alternative for distemper diagnosis, as there was a strong correlation between Ct values of conjunctival swabs and tissues ( r s = -0.8498, p < 0.00001, n = 46). This information will aid in more efficient and accurate diagnoses in individuals, small-scale outbreaks, and epidemiologic investigations in wildlife.

  6. 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.

  7. Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis.

    Science.gov (United States)

    Fischer, Cristine Dossin Bastos; Ikuta, Nilo; Canal, Cláudio Wageck; Makiejczuk, Aline; Allgayer, Mariangela da Costa; Cardoso, Cristine Hoffmeister; Lehmann, Fernanda Kieling; Fonseca, André Salvador Kazantzi; Lunge, Vagner Ricardo

    2013-12-01

    Canine distemper virus (CDV) is the cause of a severe and highly contagious disease in dogs. Practical diagnosis of canine distemper based on clinical signs and laboratory tests are required to confirm CDV infection. The present study aimed to develop a molecular assay to detect and differentiate field and vaccine CDV strains. Reverse transcription followed by nested real time polymerase chain reaction (RT-nqPCR) was developed, which exhibited analytical specificity (all the samples from healthy dogs and other canine infectious agents were not incorrectly detected) and sensitivity (all replicates of a vaccine strain were positive up to the 3125-fold dilution - 10(0.7) TCID50). RT-nqPCR was validated for CDV detection on different clinical samples (blood, urine, rectal and conjunctival swabs) of 103 animals suspected to have distemper. A total of 53 animals were found to be positive based on RT-nqPCR in at least one clinical sample. Blood resulted in more positive samples (50 out of 53, 94.3%), followed by urine (44/53, 83.0%), rectal (38/53, 71%) and conjunctival (27/53, 50.9%) swabs. A commercial immunochromatography (IC) assay had detected CDV in only 30 conjunctival samples of these positive dogs. Nucleoprotein (NC) gene sequencing of 25 samples demonstrated that 23 of them were closer to other Brazilian field strains and the remaining two to vaccine strains. A single nucleotide sequences difference, which creates an Msp I restriction enzyme digestion, was used to differentiate between field and vaccine CDV strains by restriction fragment length polymorphism (RFLP) analysis. The complete assay was more sensitive than was IC for the detection of CDV. Blood was the more frequently positive specimen and the addition of a restriction enzyme step allowed the differentiation of vaccine and Brazilian field strains. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Development of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor receptor and insulin receptor expression in equine tissue

    Directory of Open Access Journals (Sweden)

    Stephen B. Hughes

    2013-08-01

    Full Text Available The insulin-like growth factor system (insulin-like growth factor 1, insulin-like growth factor 2, insulin-like growth factor 1 receptor, insulin-like growth factor 2 receptor and six insulin-like growth factor-binding proteins and insulin are essential to muscle metabolism and most aspects of male and female reproduction. Insulin-like growth factor and insulin play important roles in the regulation of cell growth, differentiation and the maintenance of cell differentiation in mammals. In order to better understand the local factors that regulate equine physiology, such as muscle metabolism and reproduction (e.g., germ cell development and fertilisation, real-time reverse transcription polymerase chain reaction assays for quantification of equine insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were developed. The assays were sensitive: 192 copies/µLand 891 copies/µL for insulin-like growth factor 1 receptor, messenger ribonucleic acid and insulin receptor respectively (95%limit of detection, and efficient: 1.01 for the insulin-like growth factor 1 receptor assay and 0.95 for the insulin receptor assay. The assays had a broad linear range of detection (seven logs for insulin-like growth factor 1 receptor and six logs for insulin receptor. This allowed for analysis of very small amounts of messenger ribonucleic acid. Low concentrations of both insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were detected in endometrium, lung and spleen samples, whilst high concentrations were detected in heart, muscle and kidney samples, this was most likely due to the high level of glucose metabolism and glucose utilisation by these tissues. The assays developed for insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid expression have been shown to work on equine tissue and will contribute to the understanding of insulin and insulin-like growth factor 1

  9. Comparative Evaluation of Three Homogenization Methods for Isolating Middle East Respiratory Syndrome Coronavirus Nucleic Acids From Sputum Samples for Real-Time Reverse Transcription PCR.

    Science.gov (United States)

    Sung, Heungsup; Yong, Dongeun; Ki, Chang Seok; Kim, Jae Seok; Seong, Moon Woo; Lee, Hyukmin; Kim, Mi Na

    2016-09-01

    Real-time reverse transcription PCR (rRT-PCR) of sputum samples is commonly used to diagnose Middle East respiratory syndrome coronavirus (MERS-CoV) infection. Owing to the difficulty of extracting RNA from sputum containing mucus, sputum homogenization is desirable prior to nucleic acid isolation. We determined optimal homogenization methods for isolating viral nucleic acids from sputum. We evaluated the following three sputum-homogenization methods: proteinase K and DNase I (PK-DNase) treatment, phosphate-buffered saline (PBS) treatment, and N-acetyl-L-cysteine and sodium citrate (NALC) treatment. Sputum samples were spiked with inactivated MERS-CoV culture isolates. RNA was extracted from pretreated, spiked samples using the easyMAG system (bioMérieux, France). Extracted RNAs were then subjected to rRT-PCR for MERS-CoV diagnosis (DiaPlex Q MERS-coronavirus, SolGent, Korea). While analyzing 15 spiked sputum samples prepared in technical duplicate, false-negative results were obtained with five (16.7%) and four samples (13.3%), respectively, by using the PBS and NALC methods. The range of threshold cycle (Ct) values observed when detecting upE in sputum samples was 31.1-35.4 with the PK-DNase method, 34.7-39.0 with the PBS method, and 33.9-38.6 with the NALC method. Compared with the control, which were prepared by adding a one-tenth volume of 1:1,000 diluted viral culture to PBS solution, the ranges of Ct values obtained by the PBS and NALC methods differed significantly from the mean control Ct of 33.2 (both Phomogenizing sputum samples prior to RNA extraction.

  10. Preliminary results on ghrelin mRNA quantification in buffalo calves during fasting and refeeding by real-time reverse transcription PCR assay

    Directory of Open Access Journals (Sweden)

    G. Neglia

    2010-02-01

    Full Text Available The aim of this trial was to evaluate ghrelin response to milk administration in 20 days old buffalo calves. The trial was carried out on 5 female buffalo calves with a mean age of 21.2±2.8 days. Five blood samples were collected from each animal into EDTA tubes, starting at 07.00 until 15.00, at 2-h intervals. At 09.00, after the second blood sample, replaced milk was administered to the calves. Blood samples were immediately placed at 4°C until processing, which was performed on the same day. We used real-time reverse transcription PCR system to detect the expression of ghrelin mRNA levels in blood of buffalo calves. Two calves showed a low ghrelin concentration at the start of the trial (Group A = low ghrelin concentration and three calves a high ghrelin concentration (Group B = high ghrelin concentration. Ghrelin expression was significantly higher either two hours (P<0.01 and just before feeding (P<0.05 in Group B vs. Group A. However, in both cases, a significant (P<0.05 difference was observed within each group between -2 and 6 hours after feeding. Therefore, ghrelin concentration tended to increase in animals that showed low levels and, similarly, it lowered in animals that showed high concentration. If these results will be confirmed, may represent the evidence that also in buffalo calves the ghrelin system may affect feed intake. Further studies are needed in order to better evaluate the ghrelin system in buffalo calves.

  11. Classical swine fever virus detection: results of a real-time reverse transcription polymerase chain reaction ring trial conducted in the framework of the European network of excellence for epizootic disease diagnosis and control

    DEFF Research Database (Denmark)

    Hoffmann, Bernd; Blome, Sandra; Bonilauri, Paolo

    2011-01-01

    and specificity values. Nevertheless, some in-house systems had unspecific reactions or suboptimal sensitivity with only a single CSFV genotype. Follow-up actions involved either improvement of suboptimal assays or replacement of specific laboratory assays with the FLI protocol, with or without modifications......The current study reports on a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) ring trial for the detection of Classical swine fever virus (CSFV) genomic RNA undertaken by 10 European laboratories. All laboratories were asked to use their routine in-house real-time RT...

  12. Identification and validation of quantitative real-time reverse transcription PCR reference genes for gene expression analysis in teak (Tectona grandis L.f.).

    Science.gov (United States)

    Galeano, Esteban; Vasconcelos, Tarcísio Sales; Ramiro, Daniel Alves; De Martin, Valentina de Fátima; Carrer, Helaine

    2014-07-22

    Teak (Tectona grandis L.f.) is currently the preferred choice of the timber trade for fabrication of woody products due to its extraordinary qualities and is widely grown around the world. Gene expression studies are essential to explore wood formation of vascular plants, and quantitative real-time reverse transcription PCR (qRT-PCR) is a sensitive technique employed for quantifying gene expression levels. One or more appropriate reference genes are crucial to accurately compare mRNA transcripts through different tissues/organs and experimental conditions. Despite being the focus of some genetic studies, a lack of molecular information has hindered genetic exploration of teak. To date, qRT-PCR reference genes have not been identified and validated for teak. Identification and cloning of nine commonly used qRT-PCR reference genes from teak, including ribosomal protein 60s (rp60s), clathrin adaptor complexes medium subunit family (Cac), actin (Act), histone 3 (His3), sand family (Sand), β-Tubulin (Β-Tub), ubiquitin (Ubq), elongation factor 1-α (Ef-1α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Expression profiles of these genes were evaluated by qRT-PCR in six tissue and organ samples (leaf, flower, seedling, root, stem and branch secondary xylem) of teak. Appropriate gene cloning and sequencing, primer specificity and amplification efficiency was verified for each gene. Their stability as reference genes was validated by NormFinder, BestKeeper, geNorm and Delta Ct programs. Results obtained from all programs showed that TgUbq and TgEf-1α are the most stable genes to use as qRT-PCR reference genes and TgAct is the most unstable gene in teak. The relative expression of the teak cinnamyl alcohol dehydrogenase (TgCAD) gene in lignified tissues at different ages was assessed by qRT-PCR, using TgUbq and TgEf-1α as internal controls. These analyses exposed a consistent expression pattern with both reference genes. This study proposes a first broad

  13. Reverse transcription quantitative real-time polymerase chain reaction reference genes in the spared nerve injury model of neuropathic pain: validation and literature search.

    Science.gov (United States)

    Piller, Nicolas; Decosterd, Isabelle; Suter, Marc R

    2013-07-10

    The reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a widely used, highly sensitive laboratory technique to rapidly and easily detect, identify and quantify gene expression. Reliable RT-qPCR data necessitates accurate normalization with validated control genes (reference genes) whose expression is constant in all studied conditions. This stability has to be demonstrated.We performed a literature search for studies using quantitative or semi-quantitative PCR in the rat spared nerve injury (SNI) model of neuropathic pain to verify whether any reference genes had previously been validated. We then analyzed the stability over time of 7 commonly used reference genes in the nervous system - specifically in the spinal cord dorsal horn and the dorsal root ganglion (DRG). These were: Actin beta (Actb), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal proteins 18S (18S), L13a (RPL13a) and L29 (RPL29), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and hydroxymethylbilane synthase (HMBS). We compared the candidate genes and established a stability ranking using the geNorm algorithm. Finally, we assessed the number of reference genes necessary for accurate normalization in this neuropathic pain model. We found GAPDH, HMBS, Actb, HPRT1 and 18S cited as reference genes in literature on studies using the SNI model. Only HPRT1 and 18S had been once previously demonstrated as stable in RT-qPCR arrays. All the genes tested in this study, using the geNorm algorithm, presented gene stability values (M-value) acceptable enough for them to qualify as potential reference genes in both DRG and spinal cord. Using the coefficient of variation, 18S failed the 50% cut-off with a value of 61% in the DRG. The two most stable genes in the dorsal horn were RPL29 and RPL13a; in the DRG they were HPRT1 and Actb. Using a 0.15 cut-off for pairwise variations we found that any pair of stable reference gene was sufficient for the normalization process

  14. Detection of Tumor Cell-Specific mRNA in the Peripheral Blood of Patients with Breast Cancer — Evaluation of Several Markers with Real-Time Reverse Transcription-PCR

    Directory of Open Access Journals (Sweden)

    Ulrich Andergassen

    2013-01-01

    Full Text Available It is widely known that cells from epithelial tumors, e.g., breast cancer, detach from their primary tissue and enter blood circulation. We show that the presence of circulating tumor cells (CTCs in samples of patients with primary and metastatic breast cancer can be detected with an array of selected tumor-marker-genes by reverse transcription real-time PCR. The focus of the presented work is on detecting differences in gene expression between healthy individuals and adjuvant and metastatic breast cancer patients, not an accurate quantification of these differences. Therefore, total RNA was isolated from blood samples of healthy donors and patients with primary or metastatic breast cancer after enrichment of mononuclear cells by density gradient centrifugation. After reverse transcription real-time PCR was carried out with a set of marker genes (BCSP, CK8, Her2, MGL, CK18, CK19. B2M and GAPDH were used as reference genes. Blood samples from patients with metastatic disease revealed increased cytokine gene levels in comparison to normal blood samples. Detection of a single gene was not sufficient to detect CTCs by reverse transcription real-time PCR. Markers used here were selected based on a recent study detecting cancer cells on different protein levels. The combination of such a marker array leads to higher and more specific discovery rates, predominantly in metastatic patients. Identification of CTCs by PCR methods may lead to better diagnosis and prognosis and could help to choose an adequate therapy.

  15. Evaluation of a real-time PCR assay based on the single-copy SAG1 gene for the detection of Toxoplasma gondii.

    Science.gov (United States)

    Yu, Haijie; Huang, Bin; Zhuo, Xunhui; Chen, Xueqiu; Du, Aifang

    2013-11-08

    Real-time PCR-based detection of Toxoplasma gondii is very sensitive and convenient for diagnosing toxoplasmosis. However, the performance of the PCR assays could be influenced by the target gene chosen. Here we evaluate a real-time PCR assay using double-stranded DNA dyes (SYBR(®) Green I assay) with a new set of primers targeting the SAG1 gene for the fast and specific detection of T. gondii. The assay showed higher sensitivity than conventional PCR protocols using T. gondii DNA as template. The detection limit of the developed real-time PCR assay was in the order of 1 tachyzoite. The assay was also assessed by experimentally infected mice and showed positive results for blood (25%), spleen (50%) and lung (50%) as early as 1 dpi. The specificity of the assay was confirmed by using DNA from Neospora caninum, Escherichia coli, Babesia bovis, Trypanosoma brucei, Cryptosporidium parvum, and Toxocara canis. Assay applicability was successfully tested in blood samples collected from slaughtered pigs. These results indicate that, based on SYBR(®) green I, the quantitative SAG1 assay may also be useful in the study of the pathogenicity, immunoprophylaxis, and treatment of T. gondii. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Detection of live Salmonella enterica in fresh-cut vegetables by a TaqMan-based one-step reverse transcription real-time PCR.

    Science.gov (United States)

    Miao, Y J; Xiong, G T; Bai, M Y; Ge, Y; Wu, Z F

    2018-05-01

    Fresh-cut produce is at greater risk of Salmonella contamination. Detection and early warning systems play an important role in reducing the dissemination of contaminated products. One-step Reverse Transcription Polymerase Chain Reaction (RT-qPCR) targeting Salmonella tmRNA with or without a 6-h enrichment was evaluated for the detection of Salmonella in fresh-cut vegetables after 6-h storage. LOD of one-step RT-qPCR was 1·0 CFU per ml (about 100 copies tmRNA per ml) by assessed 10-fold serially diluted RNA from 10 6 CFU per ml bacteria culture. Then, one-step RT-qPCR assay was applied to detect viable Salmonella cells in 14 fresh-cut vegetables after 6-h storage. Without enrichment, this assay could detect 10 CFU per g for fresh-cut lettuce, cilantro, spinach, cabbage, Chinese cabbage and bell pepper, and 10 2 CFU per g for other vegetables. With a 6-h enrichment, this assay could detect 10 CFU per g for all fresh-cut vegetables used in this study. Moreover, this assay was able to discriminate viable cells from dead cells. This rapid detection assay may provide potential processing control and early warning method in fresh-cut vegetable processing to strengthen food safety assurance. Significance and Impact of the Study: Fresh-cut produce is at greater risk of Salmonella contamination. Rapid detection methods play an important role in reducing the dissemination of contaminated products. One-step RT-qPCR assay used in this study could detect 10 CFU per g Salmonella for 14 fresh-cut vegetables with a 6-h short enrichment. Moreover, this assay was able to discriminate viable cells from dead cells. This rapid detection assay may provide potential processing control and early warning method in fresh-cut vegetable processing to strengthen food safety assurance. © 2018 The Society for Applied Microbiology.

  17. A Novel Duplex Real-Time Reverse-Transcription PCR Assay for the Detection of Influenza A and the Novel Influenza A(H1N1 Strain

    Directory of Open Access Journals (Sweden)

    Theo P. Sloots

    2009-12-01

    Full Text Available Timely implementation of antiviral treatment and other public health based responses are dependent on accurate and rapid diagnosis of the novel pandemic influenza A(H1N1 strain. In this study we developed a duplex real-time PCR (RT-PCR (dFLU-TM assay for the simultaneous detection of a broad range of influenza A subtypes and specific detection of the novel H1N1 2009 pandemic strain. The assay was compared to the combined results of two previously described monoplex RT-PCR assays using 183 clinical samples and 10 seasonal influenza A isolates. Overall, the results showed that the dFLU-TM RT-PCR method is suitable for detection of influenza A, including the novel H1N1 pandemic strain, in clinical samples.

  18. Real-time quantitative reverse transcription-PCR analysis of expression stability of Actinobacillus pleuropneumoniae housekeeping genes during in vitro growth under iron-depleted conditions

    DEFF Research Database (Denmark)

    Nielsen, K. K.; Boye, Mette

    2005-01-01

    up-regulation under iron-restricted conditions compared to bacteria grown in medium with sufficient iron. The observed expression patterns of the genes of interest were consistent with previous observations. This study therefore lends further support to the use of real-time quantitative RT...... controls, as such controls have not been defined yet for this bacterium. Bacterial gene expression was studied during in vitro exponential and early stationary growth in medium with and without sufficient iron, respectively. First, the stability of expression of five genes, the glyA, tpiA, pykA, rec......F, and rhoAP genes involved in basic housekeeping, was evaluated on the basis of the mean pairwise variation. All the housekeeping genes included were stably expressed under the conditions investigated and consequently were included in the normalization procedure. Next, the geometric mean of the internal...

  19. Sensitivity and specificity of real-time reverse transcription polymerase chain reaction, histopathology, and immunohistochemical labeling for the detection of Rift Valley fever virus in naturally infected cattle and sheep.

    Science.gov (United States)

    Odendaal, Lieza; Fosgate, Geoffrey T; Romito, Marco; Coetzer, Jacobus A W; Clift, Sarah J

    2014-01-01

    Real-time reverse transcription polymerase chain reaction (real-time RT-PCR), histopathology, and immunohistochemical labeling (IHC) were performed on liver specimens from 380 naturally infected cattle and sheep necropsied during the 2010 Rift Valley fever (RVF) epidemic in South Africa. Sensitivity (Se) and specificity (Sp) of real-time RT-PCR, histopathology, and IHC were estimated in a latent-class model using a Bayesian framework. The Se and Sp of real-time RT-PCR were estimated as 97.4% (95% confidence interval [CI] = 95.2-98.8%) and 71.7% (95% CI = 65-77.9%) respectively. The Se and Sp of histopathology were estimated as 94.6% (95% CI = 91-97.2%) and 92.3% (95% CI = 87.6-95.8%), respectively. The Se and Sp of IHC were estimated as 97.6% (95% CI = 93.9-99.8%) and 99.4% (95% CI = 96.9-100%), respectively. Decreased Sp of real-time RT-PCR was ascribed to cross-contamination of samples. Stratified analysis of the data suggested variations in test accuracy with fetuses and severely autolyzed specimens. The Sp of histopathology in fetuses (83%) was 9.3% lower than the sample population (92.3%). The Se of IHC decreased from 97.6% to 81.5% in the presence of severe autolysis. The diagnostic Se and Sp of histopathology was higher than expected, confirming the value of routine postmortem examinations and histopathology of liver specimens. Aborted fetuses, however, should be screened using a variety of tests in areas endemic for RVF, and results from severely autolyzed specimens should be interpreted with caution. The most feasible testing option for countries lacking suitably equipped laboratories seems to be routine histology in combination with IHC.

  20. Evaluation of the PrimerDesign™ genesig real-time reverse transcription-polymerase chain reaction assay and the INFINITI® Respiratory Viral Panel Plus assay for the detection of human metapneumovirus in Kuwait.

    Science.gov (United States)

    Al-Turab, Mariam; Chehadeh, Wassim; Al-Mulla, Fahd; Al-Nakib, Widad

    2012-04-01

    Human metapneumovirus (hMPV) is a respiratory pathogen that was discovered in 2001 and is considered a major cause of both upper and lower respiratory tract infections. A sensitive, fast, and high-throughput diagnostic test is needed for the detection of hMPV that may assist in the clinical management as well as in the reduction of inappropriate therapy. Therefore, a comparison assessment was performed in this study between the PrimerDesign™ genesig real-time reverse transcription-polymerase chain reaction (RT-PCR) Assay and the INFINITI(®) Respiratory Viral Panel Plus Assay (RVP-Plus) for the detection of hMPV infection in patients with respiratory tract infections. A total of 200 respiratory samples were collected from 185 hospitalized patients, during the winter season in Kuwait. Of 185 patients, 10 (5.4%) were positive for hMPV RNA by the in-house RT-PCR assay, while 7 (4%) were positive for hMPV RNA by the real-time RT-PCR assay and 9 (5%) were positive for hMPV RNA by the INFINITI(®) RVP-Plus assay. The high incidence rate (60%) of hMPV infection was in January 2011. The sensitivity of the real-time RT-PCR and INFINITI(®) RVP-Plus assays was 70% and 90%, respectively, with specificity of 100% for both assays. hMPV types A and B could be identified in this study; however, discordant genotyping results were found between the direct sequencing method and the INFINITI(®) RVP-Plus assay in 33% of hMPV-positive patients. Copyright © 2012 Elsevier Inc. All rights reserved.

  1. Detection of African swine fever, classical swine fever, and foot-and-mouth disease viruses in swine oral fluids by multiplex reverse transcription real-time polymerase chain reaction.

    Science.gov (United States)

    Grau, Frederic R; Schroeder, Megan E; Mulhern, Erin L; McIntosh, Michael T; Bounpheng, Mangkey A

    2015-03-01

    African swine fever (ASF), classical swine fever (CSF), and foot-and-mouth disease (FMD) are highly contagious animal diseases of significant economic importance. Pigs infected with ASF and CSF viruses (ASFV and CSFV) develop clinical signs that may be indistinguishable from other diseases. Likewise, various causes of vesicular disease can mimic clinical signs caused by the FMD virus (FMDV). Early detection is critical to limiting the impact and spread of these disease outbreaks, and the ability to perform herd-level surveillance for all 3 diseases rapidly and cost effectively using a single diagnostic sample and test is highly desirable. This study assessed the feasibility of simultaneous ASFV, CSFV, and FMDV detection by multiplex reverse transcription real-time polymerase chain reaction (mRT-qPCR) in swine oral fluids collected through the use of chewing ropes. Animal groups were experimentally infected independently with each virus, observed for clinical signs, and oral fluids collected and tested throughout the course of infection. All animal groups chewed on the ropes readily before and after onset of clinical signs and before onset of lameness or serious clinical signs. ASFV was detected as early as 3 days postinoculation (dpi), 2-3 days before onset of clinical disease; CSFV was detected at 5 dpi, coincident with onset of clinical disease; and FMDV was detected as early as 1 dpi, 1 day before the onset of clinical disease. Equivalent results were observed in 4 independent studies and demonstrate the feasibility of oral fluids and mRT-qPCR for surveillance of ASF, CSF, and FMD in swine populations. © 2015 The Author(s).

  2. 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.

  3. Properties of the reverse transcription reaction in mRNA quantification

    DEFF Research Database (Denmark)

    Ståhlberg, Anders; Håkansson, Joakim; Xian, Xiaojie

    2004-01-01

    BACKGROUND: In most measurements of gene expression, mRNA is first reverse-transcribed into cDNA. We studied the reverse transcription reaction and its consequences for quantitative measurements of gene expression. METHODS: We used SYBR green I-based quantitative real-time PCR (QPCR) to measure...... the properties of reverse transcription reaction for the beta-tubulin, glyceraldehyde-3-phosphate dehydrogenase, Glut2, CaV1D, and insulin II genes, using random hexamers, oligo(dT), and gene-specific reverse transcription primers. RESULTS: Experimental variation in reverse transcription-QPCR (RT......-QPCR) was mainly attributable to the reverse transcription step. Reverse transcription efficiency depended on priming strategy, and the dependence was different for the five genes studied. Reverse transcription yields also depended on total RNA concentration. CONCLUSIONS: RT-QPCR gene expression measurements...

  4. Semi-Nested Real-Time Reverse Transcription Polymerase Chain Reaction Methods for the Successful Quantitation of Cytokeratin mRNA Expression Levels for the Subtyping of Non-Small-Cell Lung Carcinoma Using Paraffin-Embedded and Microdissected Lung Biopsy Specimens

    International Nuclear Information System (INIS)

    Nakanishi, Yoko; Shimizu, Tetsuo; Tsujino, Ichiro; Obana, Yukari; Seki, Toshimi; Fuchinoue, Fumi; Ohni, Sumie; Oinuma, Toshinori; Kusumi, Yoshiaki; Yamada, Tsutomu; Takahashi, Noriaki; Hashimoto, Shu; Nemoto, Norimichi

    2013-01-01

    In patients with inoperable advanced non-small cell lung carcinomas (NSCLCs), histological subtyping using small-mount biopsy specimens was often required to decide the indications for drug treatment. The aim of this study was to assess the utility of highly sensitive mRNA quantitation for the subtyping of advanced NSCLC using small formalin fixing and paraffin embedding (FFPE) biopsy samples. Cytokeratin (CK) 6, CK7, CK14, CK18, and thyroid transcription factor (TTF)-1 mRNA expression levels were measured using semi-nested real-time quantitative (snq) reverse-transcribed polymerase chain reaction (RT-PCR) in microdissected tumor cells collected from 52 lung biopsies. Our results using the present snqRT-PCR method showed an improvement in mRNA quantitation from small FFPE samples, and the mRNA expression level using snqRT-PCR was correlated with the immunohistochemical protein expression level. CK7, CK18, and TTF-1 mRNA were expressed at significantly higher levels (P<0.05) in adenocarcinoma (AD) than in squamous cell carcinoma (SQ), while CK6 and CK14 mRNA expression was significantly higher (P<0.05) in SQ than in AD. Each histology-specific CK, particularly CK18 in AD and CK6 in SQ, were shown to be correlated with a poor prognosis (P=0.02, 0.02, respectively). Our results demonstrated that a quantitative CK subtype mRNA analysis from lung biopsy samples can be useful for predicting the histology subtype and prognosis of advanced NSCLC

  5. Real-time systems

    OpenAIRE

    Badr, Salah M.; Bruztman, Donald P.; Nelson, Michael L.; Byrnes, Ronald Benton

    1992-01-01

    This paper presents an introduction to the basic issues involved in real-time systems. Both real-time operating sys and real-time programming languages are explored. Concurrent programming and process synchronization and communication are also discussed. The real-time requirements of the Naval Postgraduate School Autonomous Under Vehicle (AUV) are then examined. Autonomous underwater vehicle (AUV), hard real-time system, real-time operating system, real-time programming language, real-time sy...

  6. Single-copy insertion of transgenes in Caenorhabditis elegans

    DEFF Research Database (Denmark)

    Frøkjaer-Jensen, Christian; Davis, M Wayne; Hopkins, Christopher E

    2008-01-01

    developed a method that inserts a single copy of a transgene into a defined site. Mobilization of a Mos1 transposon generates a double-strand break in noncoding DNA. The break is repaired by copying DNA from an extrachromosomal template into the chromosomal site. Homozygous single-copy insertions can...... be obtained in less than 2 weeks by injecting approximately 20 worms. We have successfully inserted transgenes as long as 9 kb and verified that single copies are inserted at the targeted site. Single-copy transgenes are expressed at endogenous levels and can be expressed in the female and male germlines....

  7. Comparison of multiplex reverse transcription-PCR-enzyme ...

    African Journals Online (AJOL)

    Comparison of multiplex reverse transcription-PCR-enzyme hybridization assay with immunofluorescence techniques for the detection of four viral respiratory pathogens in pediatric community acquired pneumonia.

  8. Real Time Revisited

    Science.gov (United States)

    Allen, Phillip G.

    1985-12-01

    The call for abolishing photo reconnaissance in favor of real time is once more being heard. Ten years ago the same cries were being heard with the introduction of the Charge Coupled Device (CCD). The real time system problems that existed then and stopped real time proliferation have not been solved. The lack of an organized program by either DoD or industry has hampered any efforts to solve the problems, and as such, very little has happened in real time in the last ten years. Real time is not a replacement for photo, just as photo is not a replacement for infra-red or radar. Operational real time sensors can be designed only after their role has been defined and improvements made to the weak links in the system. Plodding ahead on a real time reconnaissance suite without benefit of evaluation of utility will allow this same paper to be used ten years from now.

  9. Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin

    Directory of Open Access Journals (Sweden)

    Kashanchi Fatah

    2006-01-01

    Full Text Available Abstract Background The early events of the HIV-1 life cycle include entry of the viral core into target cell, assembly of the reverse transcription complex (RTCs performing reverse transcription, its transformation into integration-competent complexes called pre-integration complexes (PICs, trafficking of complexes into the nucleus, and finally integration of the viral DNA into chromatin. Molecular details and temporal organization of these processes remain among the least investigated and most controversial problems in the biology of HIV. Results To quantitatively evaluate maturation and nuclear translocation of the HIV-1 RTCs, nucleoprotein complexes isolated from the nucleus (nRTC and cytoplasm (cRTC of HeLa cells infected with MLV Env-pseudotyped HIV-1 were analyzed by real-time PCR. While most complexes completed reverse transcription in the cytoplasm, some got into the nucleus before completing DNA synthesis. The HIV-specific RNA complexes could get into the nucleus when reverse transcription was blocked by reverse transcriptase inhibitor, although nuclear import of RNA complexes was less efficient than of DNA-containing RTCs. Analysis of the RTC nuclear import in synchronized cells infected in the G2/M phase of the cell cycle showed enrichment in the nuclei of RTCs containing incomplete HIV-1 DNA compared to non-synchronized cells, where RTCs with complete reverse transcripts prevailed. Immunoprecipitation assays identified viral proteins IN, Vpr, MA, and cellular Ini1 and PML associated with both cRTCs and nRTCs, whereas CA was detected only in cRTCs and RT was diminished in nRTCs. Cytoplasmic maturation of the complexes was associated with increased immunoreactivity with anti-Vpr and anti-IN antibodies, and decreased reactivity with antibodies to RT. Both cRTCs and nRTCs carried out endogenous reverse transcription reaction in vitro. In contrast to cRTCs, in vitro completion of reverse transcription in nRTCs did not increase their

  10. 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....

  11. 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

  12. Detection of enterovirus 71 gene from clinical specimens by reverse-transcription loop-mediated isothermal amplification.

    Science.gov (United States)

    Wang, D; Wang, X; Geng, Y; An, C

    2014-01-01

    The objective of this study was to develop a sensitive, specific and rapid approach to diagnose hand foot and mouth disease (HFMD) for an early treatment by using loop-mediated isothermal amplification (LAMP) technique. A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for detecting EV71 virus was developed, the specificity and sensitivity of RT-LAMP was tested, and the clinical specimens was assayed by the RT-LAMP comparing with conventional reverse-transcription polymerase chain reaction (RT-PCR) and real-time PCR. A total of 116 clinical specimens from the suspected HFMD individual were detected with the RT-LAMP. The detection rate for EV71 was 56.89% by RT-LAMP, 41.38% by real-time PCR and 34.48% by RT-PCR. The minimum detection limit of RT-LAMP was 0.01 PFU, both of RT-PCR and real-time PCR was 0.1PFU. Non-cross-reactive amplification with other enteroviruses was detected in the survey reports. The effectiveness of RT-LAMP is higher than RT-PCR and real-time PCR. The protocol is easy to operate and time saving. It was not an expensive instrument, which was needed; it is an applicable method for rapid diagnosis of the disease, especially in resource-poor countries or in developing countries.

  13. Detection of Enterovirus 71 gene from clinical specimens by reverse-transcription loop-mediated isothermal amplification

    Directory of Open Access Journals (Sweden)

    D Wang

    2014-01-01

    Full Text Available Purpose : The objective of this study was to develop a sensitive, specific and rapid approach to diagnose hand foot and mouth disease (HFMD for an early treatment by using loop-mediated isothermal amplification (LAMP technique. Materials and Methods : A reverse-transcription loop-mediated isothermal amplification (RT-LAMP for detecting EV71 virus was developed, the specificity and sensitivity of RT-LAMP was tested, and the clinical specimens was assayed by the RT-LAMP comparing with conventional reverse-transcription polymerase chain reaction (RT-PCR and real-time PCR. Results : A total of 116 clinical specimens from the suspected HFMD individual were detected with the RT-LAMP. The detection rate for EV71 was 56.89% by RT-LAMP, 41.38% by real-time PCR and 34.48% by RT-PCR. The minimum detection limit of RT-LAMP was 0.01 PFU, both of RT-PCR and real-time PCR was 0.1PFU. Non-cross-reactive amplification with other enteroviruses was detected in the survey reports. Conclusions : The effectiveness of RT-LAMP is higher than RT-PCR and real-time PCR. The protocol is easy to operate and time saving. It was not an expensive instrument, which was needed; it is an applicable method for rapid diagnosis of the disease, especially in resource-poor countries or in developing countries.

  14. 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.)

  15. 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

  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…

  17. Lineage-Specific Real-Time RT-PCR for Yellow Fever Virus Outbreak Surveillance, Brazil.

    Science.gov (United States)

    Fischer, Carlo; Torres, Maria C; Patel, Pranav; Moreira-Soto, Andres; Gould, Ernest A; Charrel, Rémi N; de Lamballerie, Xavier; Nogueira, Rita Maria Ribeiro; Sequeira, Patricia C; Rodrigues, Cintia D S; Kümmerer, Beate M; Drosten, Christian; Landt, Olfert; Bispo de Filippis, Ana Maria; Drexler, Jan Felix

    2017-11-01

    The current yellow fever outbreak in Brazil prompted widespread yellow fever virus (YFV) vaccination campaigns, imposing a responsibility to distinguish between vaccine- and wild-type YFV-associated disease. We developed novel multiplex real-time reverse transcription PCRs that differentiate between vaccine and American wild-type YFV. We validated these highly specific and sensitive assays in an outbreak setting.

  18. Lineage-Specific Real-Time RT-PCR for Yellow Fever Virus Outbreak Surveillance, Brazil

    OpenAIRE

    Fischer, Carlo; Torres, Maria C.; Patel, Pranav; Moreira-Soto, Andres; Gould, Ernest A.; Charrel, Rémi N.; de Lamballerie, Xavier; Nogueira, Rita Maria Ribeiro; Sequeira, Patricia C.; Rodrigues, Cintia D.S.; Kümmerer, Beate M.; Drosten, Christian; Landt, Olfert; Bispo de Filippis, Ana Maria; Drexler, Jan Felix

    2017-01-01

    The current yellow fever outbreak in Brazil prompted widespread yellow fever virus (YFV) vaccination campaigns, imposing a responsibility to distinguish between vaccine- and wild-type YFV-associated disease. We developed novel multiplex real-time reverse transcription PCRs that differentiate between vaccine and American wild-type YFV. We validated these highly specific and sensitive assays in an outbreak setting.

  19. Detection of three porcine vesicular viruses using multiplex real-time primer-probe energy transfer

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; Aguero, M.

    2006-01-01

    Rapid identification of the etiologic agent in infected animals is important for the control of an outbreak of vesicular disease in livestock. We have in the present study developed a multiplex real-time reverse transcription-PCR, based on primer-probe energy transfer (PriProET), for simultaneous...

  20. 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

  1. Requirements for DNA strand transfer during reverse transcription in mutant HIV-1 virions

    NARCIS (Netherlands)

    Berkhout, B.; van Wamel, J.; Klaver, B.

    1995-01-01

    Retroviruses convert their RNA genome into a DNA form by means of reverse transcription. According to the current model of reverse transcription, two strand transfer reactions are needed to synthesize a full-length DNA genome. Because reverse transcription is initiated close to the 5' end of the RNA

  2. Real time falling leaves

    OpenAIRE

    Vázquez Alcocer, Pere Pau; Balsa, Marcos

    2007-01-01

    There is a growing interest in simulating natural phenomena in computer graphics applications. Animating natural scenes in real time is one of the most challenging problems due to the inherent complexity of their structure, formed by millions of geometric entities, and the interactions that happen within. An example of natural scenario that is needed for games or simulation programs are forests. Forests are difficult to render because the huge amount of geometric entities and the large amount...

  3. Real Time Strategy Language

    OpenAIRE

    Hayes, Roy; Beling, Peter; Scherer, William

    2014-01-01

    Real Time Strategy (RTS) games provide complex domain to test the latest artificial intelligence (AI) research. In much of the literature, AI systems have been limited to playing one game. Although, this specialization has resulted in stronger AI gaming systems it does not address the key concerns of AI researcher. AI researchers seek the development of AI agents that can autonomously interpret learn, and apply new knowledge. To achieve human level performance, current AI systems rely on game...

  4. 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.

  5. Single-copy entanglement in critical quantum spin chains

    International Nuclear Information System (INIS)

    Eisert, J.; Cramer, M.

    2005-01-01

    We consider the single-copy entanglement as a quantity to assess quantum correlations in the ground state in quantum many-body systems. We show for a large class of models that already on the level of single specimens of spin chains, criticality is accompanied with the possibility of distilling a maximally entangled state of arbitrary dimension from a sufficiently large block deterministically, with local operations and classical communication. These analytical results--which refine previous results on the divergence of block entropy as the rate at which maximally entangled pairs can be distilled from many identically prepared chains--are made quantitative for general isotropic translationally invariant spin chains that can be mapped onto a quasifree fermionic system, and for the anisotropic XY model. For the XX model, we provide the asymptotic scaling of ∼(1/6)log 2 (L), and contrast it with the block entropy

  6. Real-time specifications

    DEFF Research Database (Denmark)

    David, A.; Larsen, K.G.; Legay, A.

    2015-01-01

    A specification theory combines notions of specifications and implementations with a satisfaction relation, a refinement relation, and a set of operators supporting stepwise design. We develop a specification framework for real-time systems using Timed I/O Automata as the specification formalism......, with the semantics expressed in terms of Timed I/O Transition Systems. We provide constructs for refinement, consistency checking, logical and structural composition, and quotient of specifications-all indispensable ingredients of a compositional design methodology. The theory is implemented in the new tool Ecdar...

  7. 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

  8. Real time Faraday spectrometer

    Science.gov (United States)

    Smith, Jr., Tommy E.; Struve, Kenneth W.; Colella, Nicholas J.

    1991-01-01

    This invention 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.

  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. APOBEC3G inhibits elongation of HIV-1 reverse transcripts.

    Directory of Open Access Journals (Sweden)

    Kate N Bishop

    2008-12-01

    Full Text Available APOBEC3G (A3G is a host cytidine deaminase that, in the absence of Vif, restricts HIV-1 replication and reduces the amount of viral DNA that accumulates in cells. Initial studies determined that A3G induces extensive mutation of nascent HIV-1 cDNA during reverse transcription. It has been proposed that this triggers the degradation of the viral DNA, but there is now mounting evidence that this mechanism may not be correct. Here, we use a natural endogenous reverse transcriptase assay to show that, in cell-free virus particles, A3G is able to inhibit HIV-1 cDNA accumulation not only in the absence of hypermutation but also without the apparent need for any target cell factors. We find that although reverse transcription initiates in the presence of A3G, elongation of the cDNA product is impeded. These data support the model that A3G reduces HIV-1 cDNA levels by inhibiting synthesis rather than by inducing degradation.

  11. Real time spectrum analysis

    International Nuclear Information System (INIS)

    Blunden, A.; O'Prey, D.G.; Tait, W.H.

    1983-01-01

    A method is described for the separation of a composite pulse-height spectrum into its unresolved component parts, which belong to a set of measured library spectra. The method allows real-time estimation giving running estimates during acquisition of the spectrum, minimises computation space, especially for a number of parallel calculations, estimates in advance the rms errors, and produces a significance measure for the hypothesis that the composite contains only the library spectra. Least squares curve-fitting, and other methods, can be compared, with the formalism developed, allowing analytical comparison of the effect of detector energy resolution and detection efficiency. A rational basis for the choice between the various methods of spectrum analysis follows from the theory, minimising rms estimation errors. The method described is applicable for very low numbers of counts and poor resolution. (orig.)

  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 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.

  14. 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...

  15. Normalization of Reverse Transcription Quantitative PCR Data During Ageing in Distinct Cerebral Structures.

    Science.gov (United States)

    Bruckert, G; Vivien, D; Docagne, F; Roussel, B D

    2016-04-01

    Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) has become a routine method in many laboratories. Normalization of data from experimental conditions is critical for data processing and is usually achieved by the use of a single reference gene. Nevertheless, as pointed by the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines, several reference genes should be used for reliable normalization. Ageing is a physiological process that results in a decline of many expressed genes. Reliable normalization of RT-qPCR data becomes crucial when studying ageing. Here, we propose a RT-qPCR study from four mouse brain regions (cortex, hippocampus, striatum and cerebellum) at different ages (from 8 weeks to 22 months) in which we studied the expression of nine commonly used reference genes. With the use of two different algorithms, we found that all brain structures need at least two genes for a good normalization step. We propose specific pairs of gene for efficient data normalization in the four brain regions studied. These results underline the importance of reliable reference genes for specific brain regions in ageing.

  16. Multiplex, Quantitative, Reverse Transcription PCR Detection of Influenza Viruses Using Droplet Microfluidic Technology

    Directory of Open Access Journals (Sweden)

    Ravi Prakash

    2014-12-01

    Full Text Available Quantitative, reverse transcription, polymerase chain reaction (qRT-PCR is facilitated by leveraging droplet microfluidic (DMF system, which due to its precision dispensing and sample handling capabilities at microliter and lower volumes has emerged as a popular method for miniaturization of the PCR platform. This work substantially improves and extends the functional capabilities of our previously demonstrated single qRT-PCR micro-chip, which utilized a combination of electrostatic and electrowetting droplet actuation. In the reported work we illustrate a spatially multiplexed micro-device that is capable of conducting up to eight parallel, real-time PCR reactions per usage, with adjustable control on the PCR thermal cycling parameters (both process time and temperature set-points. This micro-device has been utilized to detect and quantify the presence of two clinically relevant respiratory viruses, Influenza A and Influenza B, in human samples (nasopharyngeal swabs, throat swabs. The device performed accurate detection and quantification of the two respiratory viruses, over several orders of RNA copy counts, in unknown (blind panels of extracted patient samples with acceptably high PCR efficiency (>94%. The multi-stage qRT-PCR assays on eight panel patient samples were accomplished within 35–40 min, with a detection limit for the target Influenza virus RNAs estimated to be less than 10 RNA copies per reaction.

  17. Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Wolff, Anders; Madsen, Mogens

    2012-01-01

    Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods fordetection and quantification of this pathogen are required. In this study, we have developed a reverse transcription quantitative real-time PCR(RT-qPCR) for detection a...

  18. Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.

    Directory of Open Access Journals (Sweden)

    David Warrilow

    Full Text Available Recent work suggests a role for multiple host factors in facilitating HIV-1 reverse transcription. Previously, we identified a cellular activity which increases the efficiency of HIV-1 reverse transcription in vitro. Here, we describe aspects of the activity which shed light on its function. The cellular factor did not affect synthesis of strong-stop DNA but did improve downstream DNA synthesis. The stimulatory activity was isolated by gel filtration in a single fraction of the exclusion volume. Velocity-gradient purified HIV-1, which was free of detectable RNase activity, showed poor reverse transcription efficiency but was strongly stimulated by partially purified cell proteins. Hence, the cell factor(s did not inactivate an RNase activity that might degrade the viral genomic RNA and block completion of reverse transcription. Instead, the cell factor(s enhanced first strand transfer and synthesis of late reverse transcription suggesting it stabilized the reverse transcription complex. The factor did not affect lysis of HIV-1 by Triton X-100 in the endogenous reverse transcription (ERT system, and ERT reactions with HIV-1 containing capsid mutations, which varied the biochemical stability of viral core structures and impeded reverse transcription in cells, showed no difference in the ability to be stimulated by the cell factor(s suggesting a lack of involvement of the capsid in the in vitro assay. In addition, reverse transcription products were found to be resistant to exogenous DNase I activity when the active fraction was present in the ERT assay. These results indicate that the cell factor(s may improve reverse transcription by facilitating DNA strand transfer and DNA synthesis. It also had a protective function for the reverse transcription products, but it is unclear if this is related to improved DNA synthesis.

  19. Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

    Directory of Open Access Journals (Sweden)

    Joojin Jeong

    2015-09-01

    Full Text Available The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

  20. Validated reverse transcription droplet digital PCR serves as a higher order method for absolute quantification of Potato virus Y strains.

    Science.gov (United States)

    Mehle, Nataša; Dobnik, David; Ravnikar, Maja; Pompe Novak, Maruša

    2018-05-03

    RNA viruses have a great potential for high genetic variability and rapid evolution that is generated by mutation and recombination under selection pressure. This is also the case of Potato virus Y (PVY), which comprises a high diversity of different recombinant and non-recombinant strains. Consequently, it is hard to develop reverse transcription real-time quantitative PCR (RT-qPCR) with the same amplification efficiencies for all PVY strains which would enable their equilibrate quantification; this is specially needed in mixed infections and other studies of pathogenesis. To achieve this, we initially transferred the PVY universal RT-qPCR assay to a reverse transcription droplet digital PCR (RT-ddPCR) format. RT-ddPCR is an absolute quantification method, where a calibration curve is not needed, and it is less prone to inhibitors. The RT-ddPCR developed and validated in this study achieved a dynamic range of quantification over five orders of magnitude, and in terms of its sensitivity, it was comparable to, or even better than, RT-qPCR. RT-ddPCR showed lower measurement variability. We have shown that RT-ddPCR can be used as a reference tool for the evaluation of different RT-qPCR assays. In addition, it can be used for quantification of RNA based on in-house reference materials that can then be used as calibrators in diagnostic laboratories.

  1. Design and optimization of reverse-transcription quantitative PCR experiments.

    Science.gov (United States)

    Tichopad, Ales; Kitchen, Rob; Riedmaier, Irmgard; Becker, Christiane; Ståhlberg, Anders; Kubista, Mikael

    2009-10-01

    Quantitative PCR (qPCR) is a valuable technique for accurately and reliably profiling and quantifying gene expression. Typically, samples obtained from the organism of study have to be processed via several preparative steps before qPCR. We estimated the errors of sample withdrawal and extraction, reverse transcription (RT), and qPCR that are introduced into measurements of mRNA concentrations. We performed hierarchically arranged experiments with 3 animals, 3 samples, 3 RT reactions, and 3 qPCRs and quantified the expression of several genes in solid tissue, blood, cell culture, and single cells. A nested ANOVA design was used to model the experiments, and relative and absolute errors were calculated with this model for each processing level in the hierarchical design. We found that intersubject differences became easily confounded by sample heterogeneity for single cells and solid tissue. In cell cultures and blood, the noise from the RT and qPCR steps contributed substantially to the overall error because the sampling noise was less pronounced. We recommend the use of sample replicates preferentially to any other replicates when working with solid tissue, cell cultures, and single cells, and we recommend the use of RT replicates when working with blood. We show how an optimal sampling plan can be calculated for a limited budget. .

  2. HIV-1 reverse transcription initiation: a potential target for novel antivirals?

    NARCIS (Netherlands)

    Abbink, Truus E. M.; Berkhout, Ben

    2008-01-01

    Reverse transcription is an essential step in the retroviral life cycle, as it converts the genomic RNA into DNA. In this review, we describe recent developments concerning the initiation step of this complex, multi-step reaction. During initiation of reverse transcription, a cellular tRNA primer is

  3. Improved detection limit in rapid detection of human enterovirus 71 and coxsackievirus A16 by a novel reverse transcription-isothermal multiple-self-matching-initiated amplification assay.

    Science.gov (United States)

    Ding, Xiong; Nie, Kai; Shi, Lei; Zhang, Yong; Guan, Li; Zhang, Dan; Qi, Shunxiang; Ma, Xuejun

    2014-06-01

    Rapid detection of human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) is important in the early phase of hand-foot-and-mouth disease (HFMD). In this study, we developed and evaluated a novel reverse transcription-isothermal multiple-self-matching-initiated amplification (RT-IMSA) assay for the rapid detection of EV71 and CVA16 by use of reverse transcriptase, together with a strand displacement DNA polymerase. Real-time RT-IMSA assays using a turbidimeter and visual RT-IMSA assays to detect EV71 and CVA16 were established and completed in 1 h, and the reported corresponding real-time reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assays targeting the same regions of the VP1 gene were adopted as parallel tests. Through testing VP1 RNAs transcribed in vitro, the real-time RT-IMSA assays exhibited better linearity of quantification, with R(2) values of 0.952 (for EV71) and 0.967 (for CVA16), than the real-time RT-LAMP assays, which had R(2) values of 0.803 (for EV71) and 0.904 (for CVA16). Additionally, the detection limits of the real-time RT-IMSA assays (approximately 937 for EV71 and 67 for CVA16 copies/reaction) were higher than those of real-time RT-LAMP assays (approximately 3,266 for EV71 and 430 for CVA16 copies/reaction), and similar results were observed in the visual RT-IMSA assays. The new approaches also possess high specificities for the corresponding targets, with no cross-reactivity observed. In clinical assessment, compared to commercial reverse transcription-quantitative PCR (qRT-PCR) kits, the diagnostic sensitivities of the real-time RT-IMSA assays (96.4% for EV71 and 94.6% for CVA16) were higher than those of the real-time RT-LAMP assays (91.1% for EV71 and 90.8% for CVA16). The visual RT-IMSA assays also exhibited the same results. In conclusion, this proof-of-concept study suggests that the novel RT-IMSA assay is superior to the RT-LAMP assay in terms of detection limit and has the potential to rapidly detect EV71

  4. Dependable Real-Time Systems

    Science.gov (United States)

    1991-09-30

    0196 or 413 545-0720 PI E-mail Address: krithi@nirvan.cs.umass.edu, stankovic(ocs.umass.edu Grant or Contract Title: Dependable Real - Time Systems Grant...Dependable Real - Time Systems " Grant or Contract Number: N00014-85-k-0398 L " Reporting Period: 1 Oct 87 - 30 Sep 91 , 2. Summary of Accomplishments ’ 2.1 Our...in developing a sound approach to scheduling tasks in complex real - time systems , (2) developed a real-time operating system kernel, a preliminary

  5. Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

    DEFF Research Database (Denmark)

    Dukes, J.P.; King, D.P.; Alexandersen, Søren

    2006-01-01

    Speed is paramount in the diagnosis of foot-and-mouth disease (FMD) and simplicity is required if a test is to be deployed in the field. The development of a one-step, reverse transcription loop-mediated amplification (RT-LAMP) assay enables FMD virus (FMDV) to be detected in under an hour...... in a single tube without thermal cycling. A fragment of the 3D RNA polymerase gene of the virus is amplified at 65 degrees C in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase. Compared with real-time RT-PCR, RT-LAMP was consistently faster, and ten copies of FMDV...... vesicular diseases and from that of genetically related picornaviruses. Diagnostic sensitivity was validated by the amplification of reference FMDV strains and archival material from field cases of FMD. In comparison with the performance of the established diagnostic TaqMan (R) assay, RT-LAMP appears...

  6. Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction.

    Science.gov (United States)

    Wilkes, Rebecca P; Tsai, Yun-Long; Lee, Pei-Yu; Lee, Fu-Chun; Chang, Hsiao-Fen Grace; Wang, Hwa-Tang Thomas

    2014-09-09

    Canine distemper virus (CDV) has been associated with outbreaks of canine infectious respiratory disease in shelters and boarding kennel environments. POCKITTM Nucleic Acid Analyzer is a field-deployable device capable of generating automatically interpreted insulated isothermal polymerase chain reaction (iiPCR) results from extracted nucleic acid within one hour. In this study, reverse transcription iiPCR (RT-iiPCR) was developed to facilitate point-of-need diagnosis of CDV infection. Analytical sensitivity (limit of detection 95%) of the established CDV RT-iiPCR was about 11 copies of in vitro transcribed RNA per reaction. CDV RT-iiPCR generated positive signals from CDV, but not Bordetella bronchiseptica, canine parvovirus, canine herpesvirus, canine adenovirus 2, canine influenza virus (subtype H3N8), canine parainfluenza virus, and canine respiratory coronavirus. To evaluate accuracy of the established reaction in canine distemper clinical diagnosis, 110 specimens from dogs, raccoons, and foxes suspected with CDV infection were tested simultaneously by CDV RT-iiPCR and real-time RT-PCR. CDV RT-iiPCR demonstrated excellent sensitivity (100%) and specificity (100%), compared to real-time RT-PCR. The results indicated an excellent correlation between RT-iiPCR and a reference real time RT-PCR method. Working in a lyophilized format, the established method has great potential to be used for point-of-care diagnosis of canine distemper in animals, especially in resource-limited facilities.

  7. The reverse transcription inhibitor abacavir shows anticancer activity in prostate cancer cell lines.

    Directory of Open Access Journals (Sweden)

    Francesca Carlini

    Full Text Available BACKGROUND: Transposable Elements (TEs comprise nearly 45% of the entire genome and are part of sophisticated regulatory network systems that control developmental processes in normal and pathological conditions. The retroviral/retrotransposon gene machinery consists mainly of Long Interspersed Nuclear Elements (LINEs-1 and Human Endogenous Retroviruses (HERVs that code for their own endogenous reverse transcriptase (RT. Interestingly, RT is typically expressed at high levels in cancer cells. Recent studies report that RT inhibition by non-nucleoside reverse transcriptase inhibitors (NNRTIs induces growth arrest and cell differentiation in vitro and antagonizes growth of human tumors in animal model. In the present study we analyze the anticancer activity of Abacavir (ABC, a nucleoside reverse transcription inhibitor (NRTI, on PC3 and LNCaP prostate cancer cell lines. PRINCIPAL FINDINGS: ABC significantly reduces cell growth, migration and invasion processes, considerably slows S phase progression, induces senescence and cell death in prostate cancer cells. Consistent with these observations, microarray analysis on PC3 cells shows that ABC induces specific and dose-dependent changes in gene expression, involving multiple cellular pathways. Notably, by quantitative Real-Time PCR we found that LINE-1 ORF1 and ORF2 mRNA levels were significantly up-regulated by ABC treatment. CONCLUSIONS: Our results demonstrate the potential of ABC as anticancer agent able to induce antiproliferative activity and trigger senescence in prostate cancer cells. Noteworthy, we show that ABC elicits up-regulation of LINE-1 expression, suggesting the involvement of these elements in the observed cellular modifications.

  8. 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...

  9. Prototyping real-time systems

    OpenAIRE

    Clynch, Gary

    1994-01-01

    The traditional software development paradigm, the waterfall life cycle model, is defective when used for developing real-time systems. This thesis puts forward an executable prototyping approach for the development of real-time systems. A prototyping system is proposed which uses ESML (Extended Systems Modelling Language) as a prototype specification language. The prototyping system advocates the translation of non-executable ESML specifications into executable LOOPN (Language of Object ...

  10. 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.

  11. Reference genes for reverse transcription quantitative PCR in canine brain tissue

    NARCIS (Netherlands)

    Stassen, Quirine E M; Riemers, Frank M; Reijmerink, Hannah; Leegwater, Peter A J; Penning, Louis C

    2015-01-01

    BACKGROUND: In the last decade canine models have been used extensively to study genetic causes of neurological disorders such as epilepsy and Alzheimer's disease and unravel their pathophysiological pathways. Reverse transcription quantitative polymerase chain reaction is a sensitive and

  12. Towards Real-Time Argumentation

    Directory of Open Access Journals (Sweden)

    Vicente JULIÁN

    2016-07-01

    Full Text Available In this paper, we deal with the problem of real-time coordination with the more general approach of reaching real-time agreements in MAS. Concretely, this work proposes a real-time argumentation framework in an attempt to provide agents with the ability of engaging in argumentative dialogues and come with a solution for their underlying agreement process within a bounded period of time. The framework has been implemented and evaluated in the domain of a customer support application. Concretely, we consider a society of agents that act on behalf of a group of technicians that must solve problems in a Technology Management Centre (TMC within a bounded time. This centre controls every process implicated in the provision of technological and customer support services to private or public organisations by means of a call centre. The contract signed between the TCM and the customer establishes penalties if the specified time is exceeded.

  13. 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

  14. 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...

  15. 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.

  16. Designing Real Time Assistive Technologies

    DEFF Research Database (Denmark)

    Sonne, Tobias; Obel, Carsten; Grønbæk, Kaj

    2015-01-01

    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...

  17. 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

  18. ISTTOK real-time architecture

    International Nuclear Information System (INIS)

    Carvalho, Ivo S.; 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.

    2014-01-01

    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 ® Q8200 4-core processor running at 2.33 GHz located in the ATCA crate. Two boards (inside the ATCA crate) with 32 analog

  19. 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

  20. 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

  1. 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.

  2. [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.

  3. Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Dufva, I.H.; Dufva, Hans Martin

    2006-01-01

    oligonucleotides (pentadecamers) consistently, yielded at least 2 fold as much cDNA as did random hexamers using either-poly(A) RNA or an amplified version of messenger RNA (aRNA) as a template. The cDNA generated using pentadecamers did not differ in size distribution or the amount of incorporated label compared...... with cDNA generated with random hexamers. The increased efficiency of priming using random pentadecamers resulted in reverse transcription of > 80% of the template aRNA, while random hexamers induced reverse transcription of only 40% of the template aRNA. This suggests a better coverage...... that random pentadecamers can replace random hexamers in reverse transcription reactions on both poly(A) RNA and amplified RNA, resulting in higher cDNA yields and quality....

  4. The specificity and flexibility of l1 reverse transcription priming at imperfect T-tracts.

    Directory of Open Access Journals (Sweden)

    Clément Monot

    2013-05-01

    Full Text Available L1 retrotransposons have a prominent role in reshaping mammalian genomes. To replicate, the L1 ribonucleoprotein particle (RNP first uses its endonuclease (EN to nick the genomic DNA. The newly generated DNA end is subsequently used as a primer to initiate reverse transcription within the L1 RNA poly(A tail, a process known as target-primed reverse transcription (TPRT. Prior studies demonstrated that most L1 insertions occur into sequences related to the L1 EN consensus sequence (degenerate 5'-TTTT/A-3' sites and frequently preceded by imperfect T-tracts. However, it is currently unclear whether--and to which degree--the liberated 3'-hydroxyl extremity on the genomic DNA needs to be accessible and complementary to the poly(A tail of the L1 RNA for efficient priming of reverse transcription. Here, we employed a direct assay for the initiation of L1 reverse transcription to define the molecular rules that guide this process. First, efficient priming is detected with as few as 4 matching nucleotides at the primer 3' end. Second, L1 RNP can tolerate terminal mismatches if they are compensated within the 10 last bases of the primer by an increased number of matching nucleotides. All terminal mismatches are not equally detrimental to DNA extension, a C being extended at higher levels than an A or a G. Third, efficient priming in the context of duplex DNA requires a 3' overhang. This suggests the possible existence of additional DNA processing steps, which generate a single-stranded 3' end to allow L1 reverse transcription. Based on these data we propose that the specificity of L1 reverse transcription initiation contributes, together with the specificity of the initial EN cleavage, to the distribution of new L1 insertions within the human genome.

  5. The specificity and flexibility of l1 reverse transcription priming at imperfect T-tracts.

    Science.gov (United States)

    Monot, Clément; Kuciak, Monika; Viollet, Sébastien; Mir, Ashfaq Ali; Gabus, Caroline; Darlix, Jean-Luc; Cristofari, Gaël

    2013-05-01

    L1 retrotransposons have a prominent role in reshaping mammalian genomes. To replicate, the L1 ribonucleoprotein particle (RNP) first uses its endonuclease (EN) to nick the genomic DNA. The newly generated DNA end is subsequently used as a primer to initiate reverse transcription within the L1 RNA poly(A) tail, a process known as target-primed reverse transcription (TPRT). Prior studies demonstrated that most L1 insertions occur into sequences related to the L1 EN consensus sequence (degenerate 5'-TTTT/A-3' sites) and frequently preceded by imperfect T-tracts. However, it is currently unclear whether--and to which degree--the liberated 3'-hydroxyl extremity on the genomic DNA needs to be accessible and complementary to the poly(A) tail of the L1 RNA for efficient priming of reverse transcription. Here, we employed a direct assay for the initiation of L1 reverse transcription to define the molecular rules that guide this process. First, efficient priming is detected with as few as 4 matching nucleotides at the primer 3' end. Second, L1 RNP can tolerate terminal mismatches if they are compensated within the 10 last bases of the primer by an increased number of matching nucleotides. All terminal mismatches are not equally detrimental to DNA extension, a C being extended at higher levels than an A or a G. Third, efficient priming in the context of duplex DNA requires a 3' overhang. This suggests the possible existence of additional DNA processing steps, which generate a single-stranded 3' end to allow L1 reverse transcription. Based on these data we propose that the specificity of L1 reverse transcription initiation contributes, together with the specificity of the initial EN cleavage, to the distribution of new L1 insertions within the human genome.

  6. The real time rolling shutter

    OpenAIRE

    Monaghan, David; O'Connor, Noel E.; Cleary, Anne; Connolly, Denis

    2015-01-01

    From an early age children are often told either, you are creative you should do art but stay away from science and maths. Or that you are mathematical you should do science but you're not that creative. Compounding this there also exist some traditional barriers of artistic rhetoric that say, "don't touch, don't think and don't be creative, we've already done that for you, you can just look...". The Real Time Rolling Shutter is part of a collaborative Art/Science partnership whose core tenet...

  7. Rapid Identification of Dengue Virus by Reverse Transcription-Polymerase Chain Reaction Using Field-Deployable Instrumentation

    National Research Council Canada - National Science Library

    McAvin, James C; Escamilla, Elizabeth M; Blow, James A; Turell, Micahel J; Quintana, Miguel; Bowles, David E; Swaby, James A; Barnes, William J; Huff, William B; Lahman, Kenton L

    2005-01-01

    ...) reverse transcription-polymerase chain reaction assays were developed for screening and seroype identification of infected mosquito vectors and human sera using a field-deployable, fluorometric thermocycler...

  8. Reliable gene expression analysis by reverse transcription-quantitative PCR: reporting and minimizing the uncertainty in data accuracy.

    Science.gov (United States)

    Remans, Tony; Keunen, Els; Bex, Geert Jan; Smeets, Karen; Vangronsveld, Jaco; Cuypers, Ann

    2014-10-01

    Reverse transcription-quantitative PCR (RT-qPCR) has been widely adopted to measure differences in mRNA levels; however, biological and technical variation strongly affects the accuracy of the reported differences. RT-qPCR specialists have warned that, unless researchers minimize this variability, they may report inaccurate differences and draw incorrect biological conclusions. The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines describe procedures for conducting and reporting RT-qPCR experiments. The MIQE guidelines enable others to judge the reliability of reported results; however, a recent literature survey found low adherence to these guidelines. Additionally, even experiments that use appropriate procedures remain subject to individual variation that statistical methods cannot correct. For example, since ideal reference genes do not exist, the widely used method of normalizing RT-qPCR data to reference genes generates background noise that affects the accuracy of measured changes in mRNA levels. However, current RT-qPCR data reporting styles ignore this source of variation. In this commentary, we direct researchers to appropriate procedures, outline a method to present the remaining uncertainty in data accuracy, and propose an intuitive way to select reference genes to minimize uncertainty. Reporting the uncertainty in data accuracy also serves for quality assessment, enabling researchers and peer reviewers to confidently evaluate the reliability of gene expression data. © 2014 American Society of Plant Biologists. All rights reserved.

  9. Candidate gene biodosimeters of mice and human exposure to ionizing radiation by quantitative reverse transcription polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    Hamed Rezaeejam

    2015-01-01

    Full Text Available Understanding of cellular responses to ionizing radiation (IR is essential for the development of predictive markers useful for assessing human exposure. Biological markers of exposure to IR in human populations are of great interest for assessing normal tissue injury in radiation oncology and for biodosimetry in nuclear incidents and accidental radiation exposures. Traditional radiation exposure biomarkers based on cytogenetic assays (biodosimetry, are time-consuming and do not provide results fast enough and requires highly trained personnel for scoring. Hence, the development of rapid biodosimetry methods is one of the highest priorities. Exposure of cells to IR activates multiple signal transduction pathways, which result in complex alterations in gene-expression. Real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR has become the benchmark for the detection and quantification of RNA targets and is being utilized increasingly in monitoring the specific genes with more accurately and sensitively. This review evaluates the RT-qPCR as a biodosimetry method and we investigated the papers from 2000 up to now, which identified the genes-expression related the DNA repair, cell cycle checkpoint, and apoptosis induced by ionization radiation in peripheral blood and determined as biodosimeters. In conclusion, it could be say that RT-qPCR technique for determining the specific genes as biodosimeters could be a fully quantitative reliable and sensitive method. Furthermore, the results of the current review will help the researchers to recognize the most expressed genes induced by ionization radiation.

  10. 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

  11. Structural features in the HIV-1 repeat region facilitate strand transfer during reverse transcription

    NARCIS (Netherlands)

    Berkhout, B.; Vastenhouw, N. L.; Klasens, B. I.; Huthoff, H.

    2001-01-01

    Two obligatory DNA strand transfers take place during reverse transcription of a retroviral RNA genome. The first strand transfer is facilitated by terminal repeat (R) elements in the viral genome. This strand-transfer reaction depends on base pairing between the cDNA of the 5'R and the 3'R. There

  12. Autonomous Real Time Requirements Tracing

    Science.gov (United States)

    Plattsmier, George; Stetson, Howard

    2014-01-01

    One of the more challenging aspects of software development is the ability to verify and validate the functional software requirements dictated by the Software Requirements Specification (SRS) and the Software Detail Design (SDD). Insuring the software has achieved the intended requirements is the responsibility of the Software Quality team and the Software Test team. The utilization of Timeliner-TLX(sup TM) Auto- Procedures for relocating ground operations positions to ISS automated on-board operations has begun the transition that would be required for manned deep space missions with minimal crew requirements. This transition also moves the auto-procedures from the procedure realm into the flight software arena and as such the operational requirements and testing will be more structured and rigorous. The autoprocedures would be required to meet NASA software standards as specified in the Software Safety Standard (NASASTD- 8719), the Software Engineering Requirements (NPR 7150), the Software Assurance Standard (NASA-STD-8739) and also the Human Rating Requirements (NPR-8705). The Autonomous Fluid Transfer System (AFTS) test-bed utilizes the Timeliner-TLX(sup TM) Language for development of autonomous command and control software. The Timeliner-TLX(sup TM) system has the unique feature of providing the current line of the statement in execution during real-time execution of the software. The feature of execution line number internal reporting unlocks the capability of monitoring the execution autonomously by use of a companion Timeliner-TLX(sup TM) sequence as the line number reporting is embedded inside the Timeliner-TLX(sup TM) execution engine. This negates I/O processing of this type data as the line number status of executing sequences is built-in as a function reference. This paper will outline the design and capabilities of the AFTS Autonomous Requirements Tracker, which traces and logs SRS requirements as they are being met during real-time execution of the

  13. Inter-laboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction methods used for detection of infectious pancreatic necrosis virus in Chile

    Directory of Open Access Journals (Sweden)

    David Tapia

    2017-07-01

    Conclusions: Overall, the ring trial showed high values of sensitivity and specificity, with some problems of repeatability and inter-laboratory variability. This last issue needs to be addressed in order to allow harmonized diagnostic of IPNV within the country. We recommend the use of the NRL methods as validated and reliable qRT-PCR protocols for the detection of IPNV.

  14. Evaluation of reference genes for reverse transcription quantitative real-time PCR (RT-qPCR) studies in Silene vulgaris considering the method of cDNA preparation

    Czech Academy of Sciences Publication Activity Database

    Koloušková, Pavla; Stone, James D.; Štorchová, Helena

    2017-01-01

    Roč. 12, č. 8 (2017), č. článku e0183470. E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) LH15075; GA ČR(CZ) GA16-09220S Institutional support: RVO:61389030 Keywords : CYTOPLASMIC MALE -STERILITY * SUITABLE REFERENCE GENES * INTERNAL CONTROL GENES Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 2.806, year: 2016

  15. Mobile real time radiography system

    Energy Technology Data Exchange (ETDEWEB)

    Vigil, J.; Taggart, D.; Betts, S. [Los Alamos National Lab., NM (United States)] [and others

    1997-11-01

    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 {approximately}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.

  16. 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

  17. 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.

  18. 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

  19. 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...

  20. Hard Real-Time Networking on Firewire

    NARCIS (Netherlands)

    Zhang, Yuchen; Orlic, Bojan; Visser, Peter; Broenink, Jan

    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

  1. Modular specification of real-time systems

    DEFF Research Database (Denmark)

    Inal, Recep

    1994-01-01

    Duration Calculus, a real-time interval logic, has been embedded in the Z specification language to provide a notation for real-time systems that combines the modularisation and abstraction facilities of Z with a logic suitable for reasoning about real-time properties. In this article the notation...

  2. Students Collecting Real time Data

    Science.gov (United States)

    Miller, P.

    2006-05-01

    Students Collecting Real-Time Data The Hawaiian Islands Humpback Whale National Marine Sanctuary has created opportunities for middle and high school students to become Student Researchers and to be involved in real-time marine data collection. It is important that we expose students to different fields of science and encourage them to enter scientific fields of study. The Humpback Whale Sanctuary has an education visitor center in Kihei, Maui. Located right on the beach, the site has become a living classroom facility. There is a traditional Hawaiian fishpond fronting the property. The fishpond wall is being restored, using traditional methods. The site has the incredible opportunity of incorporating Hawaiian cultural practices with scientific studies. The Sanctuary offers opportunities for students to get involved in monitoring and data collection studies. Invasive Seaweed Study: Students are collecting data on invasive seaweed for the University of Hawaii. They pull a large net through the shallow waters. Seaweed is sorted, identified and weighed. The invasive seaweeds are removed. The data is recorded and sent to UH. Remote controlled monitoring boats: The sanctuary has 6 boogie board sized remote controlled boats used to monitor reefs. Boats have a camera with lights on the underside. The boats have water quality monitoring devices and GPS units. The video from the underwater camera is transmitted via a wireless transmission. Students are able to monitor the fish, limu and invertebrate populations on the reef and collect water quality data via television monitors or computers. The boat can also pull a small plankton tow net. Data is being compiled into data bases. Artificial Reef Modules: The Sanctuary has a scientific permit from the state to build and deploy artificial reef modules. High school students are designing and building modules. These are deployed out in the Fishpond fronting the Sanctuary site and students are monitoring them on a weekly basis

  3. Generation of single-copy transgenic mouse embryos directly from ES cells by tetraploid embryo complementation

    Directory of Open Access Journals (Sweden)

    Zhao Roong

    2001-12-01

    Full Text Available Abstract Background Transgenic mice have been used extensively to analyze gene function. Unfortunately, traditional transgenic procedures have only limited use in analyzing alleles that cause lethality because lines of founder mice cannot be established. This is frustrating given that such alleles often reveal crucial aspects of gene function. For this reason techniques that facilitate the generation of embryos expressing such alleles would be of enormous benefit. Although the transient generation of transgenic embryos has allowed limited analysis of lethal alleles, it is expensive, time consuming and technically challenging. Moreover a fundamental limitation with this approach is that each embryo generated is unique and transgene expression is highly variable due to the integration of different transgene copy numbers at random genomic sites. Results Here we describe an alternative method that allows the generation of clonal mouse embryos harboring a single-copy transgene at a defined genomic location. This was facilitated through the production of Hprt negative embryonic stem cells that allow the derivation of embryos by tetraploid embryo complementation. We show that targeting transgenes to the hprt locus in these ES cells by homologous recombination can be efficiently selected by growth in HAT medium. Moreover, embryos derived solely from targeted ES cells containing a single copy LacZ transgene under the control of the α-myosin heavy chain promoter exhibited the expected cardiac specific expression pattern. Conclusion Our results demonstrate that tetraploid embryo complementation by F3 hprt negative ES cells facilitates the generation of transgenic mouse embryos containing a single copy gene at a defined genomic locus. This approach is simple, extremely efficient and bypasses any requirement to generate chimeric mice. Moreover embryos generated by this procedure are clonal in that they are all derived from a single ES cell lines. This

  4. Identification of valid reference genes for gene expression studies of human stomach cancer by reverse transcription-qPCR

    Directory of Open Access Journals (Sweden)

    Lee Yeon-Su

    2010-05-01

    Full Text Available Abstract Background Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR is a powerful method for the analysis of gene expression. Target gene expression levels are usually normalized to a consistently expressed reference gene also known as internal standard, in the same sample. However, much effort has not been expended thus far in the search for reference genes suitable for the study of stomach cancer using RT-qPCR, although selection of optimal reference genes is critical for interpretation of results. Methods We assessed the suitability of six possible reference genes, beta-actin (ACTB, glyceraldehydes-3-phosphate dehydrogenase (GAPDH, hypoxanthine phosphoribosyl transferase 1 (HPRT1, beta-2-microglobulin (B2M, ribosomal subunit L29 (RPL29 and 18S ribosomal RNA (18S rRNA in 20 normal and tumor stomach tissue pairs of stomach cancer patients and 6 stomach cancer cell lines, by RT-qPCR. Employing expression stability analyses using NormFinder and geNorm algorithms we determined the order of performance of these reference genes and their variation values. Results This RT-qPCR study showed that there are statistically significant (p Conclusion This study validated RPL29 and RPL29-B2M as the best single reference genes and combination, for RT-qPCR analysis of 'all stomach tissues', and B2M and B2M-GAPDH as the best single reference gene and combination, for 'stomach cancer cell lines'. Use of these validated reference genes should provide more exact interpretation of differential gene expressions at transcription level in stomach cancer.

  5. Microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse-transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens.

    Science.gov (United States)

    Jia, Ruan; Chengjun, Sun; Heng, Chen; Chen, Zhou; Yuanqian, Li; Yongxin, Li

    2015-07-01

    Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. VERSE - Virtual Equivalent Real-time Simulation

    Science.gov (United States)

    Zheng, Yang; Martin, Bryan J.; Villaume, Nathaniel

    2005-01-01

    Distributed real-time simulations provide important timing validation and hardware in the- loop results for the spacecraft flight software development cycle. Occasionally, the need for higher fidelity modeling and more comprehensive debugging capabilities - combined with a limited amount of computational resources - calls for a non real-time simulation environment that mimics the real-time environment. By creating a non real-time environment that accommodates simulations and flight software designed for a multi-CPU real-time system, we can save development time, cut mission costs, and reduce the likelihood of errors. This paper presents such a solution: Virtual Equivalent Real-time Simulation Environment (VERSE). VERSE turns the real-time operating system RTAI (Real-time Application Interface) into an event driven simulator that runs in virtual real time. Designed to keep the original RTAI architecture as intact as possible, and therefore inheriting RTAI's many capabilities, VERSE was implemented with remarkably little change to the RTAI source code. This small footprint together with use of the same API allows users to easily run the same application in both real-time and virtual time environments. VERSE has been used to build a workstation testbed for NASA's Space Interferometry Mission (SIM PlanetQuest) instrument flight software. With its flexible simulation controls and inexpensive setup and replication costs, VERSE will become an invaluable tool in future mission development.

  7. Space Weather and Real-Time Monitoring

    Directory of Open Access Journals (Sweden)

    S Watari

    2009-04-01

    Full Text Available Recent advance of information and communications technology enables to collect a large amount of ground-based and space-based observation data in real-time. The real-time data realize nowcast of space weather. This paper reports a history of space weather by the International Space Environment Service (ISES in association with the International Geophysical Year (IGY and importance of real-time monitoring in space weather.

  8. Archtecture of distributed real-time systems

    OpenAIRE

    Wing Leung, Cheuk

    2013-01-01

    CRAFTERS (Constraint and Application Driven Framework for Tailoring Embedded Real-time System) project aims to address the problem of uncertainty and heterogeneity in a distributed system by providing seamless, portable connectivity and middleware. This thesis contributes to the project by investigating the techniques that can be used in a distributed real-time embedded system. The conclusion is that, there is a list of specifications to be meet in order to provide a transparent and real-time...

  9. Research Directions in Real-Time Systems.

    Science.gov (United States)

    1996-09-01

    This report summarizes a survey of published research in real time systems . Material is presented that provides an overview of the topic, focusing on...communications protocols and scheduling techniques. It is noted that real - time systems deserve special attention separate from other areas because of...formal tools for design and analysis of real - time systems . The early work on applications as well as notable theoretical advances are summarized

  10. A Real-Time Systems Symposium Preprint.

    Science.gov (United States)

    1983-09-01

    Real - Time Systems Symposium Preprint Interim Tech...estimate of the occurence of the error. Unclassii ledSECUqITY CLASSIF’ICA T" NO MI*IA If’ inDI /’rrd erter for~~ble. ’Corrputnqg A REAL - TIME SYSTEMS SYMPOSIUM...ABSTRACT This technical report contains a preprint of a paper accepted for presentation at the REAL - TIME SYSTEMS SYMPOSIUM, Arlington,

  11. Essays in real-time forecasting

    OpenAIRE

    Liebermann, Joelle

    2012-01-01

    This thesis contains three essays in the field of real-time econometrics, and more particularlyforecasting.The issue of using data as available in real-time to forecasters, policymakers or financialmarkets is an important one which has only recently been taken on board in the empiricalliterature. Data available and used in real-time are preliminary and differ from ex-postrevised data, and given that data revisions may be quite substantial, the use of latestavailable instead of real-time can s...

  12. 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.

  13. 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...

  14. 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...

  15. Sequence-specific inhibition of duck hepatitis B virus reverse transcription by peptide nucleic acids (PNA)

    DEFF Research Database (Denmark)

    Robaczewska, Magdalena; Narayan, Ramamurthy; Seigneres, Beatrice

    2005-01-01

    BACKGROUND/AIMS: Peptide nucleic acids (PNAs) appear as promising new antisense agents, that have not yet been examined as hepatitis B virus (HBV) inhibitors. Our aim was to study the ability of PNAs targeting the duck HBV (DHBV) encapsidation signal epsilon to inhibit reverse transcription (RT...... in primary duck hepatocytes (PDH). RESULTS: Both PNAs reproducibly inhibited DHBV RT in a dose-dependent manner with IC(50) of 10nM, whereas up to 600-fold higher concentration of S-ODNs was required for similar inhibition. The PNA targeting the bulge and upper stem of epsilon appeared as more efficient RT...

  16. 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

  17. Towards exascale real-time RFI mitigation

    NARCIS (Netherlands)

    van Nieuwpoort, R.V.

    2016-01-01

    We describe the design and implementation of an extremely scalable real-time RFI mitigation method, based on the offline AOFlagger. All algorithms scale linearly in the number of samples. We describe how we implemented the flagger in the LOFAR real-time pipeline, on both CPUs and GPUs. Additionally,

  18. 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

  19. De toekomst van Real Time Intelligence

    NARCIS (Netherlands)

    Broek, J. van den; Berg, C.H. van den

    2013-01-01

    Al direct vanaf de start van de Nationale Politie is gewerkt aan het opzetten van tien real-time intelligence centra in Nederland. Van daaruit worden 24 uur per dag en zeven dagen in de week agenten op straat actief ondersteund met real-time informatie bij de melding waar ze op af gaan. In de visie

  20. Heterogeneous Embedded Real-Time Systems Environment

    Science.gov (United States)

    2003-12-01

    AFRL-IF-RS-TR-2003-290 Final Technical Report December 2003 HETEROGENEOUS EMBEDDED REAL - TIME SYSTEMS ENVIRONMENT Integrated...HETEROGENEOUS EMBEDDED REAL - TIME SYSTEMS ENVIRONMENT 6. AUTHOR(S) Cosmo Castellano and James Graham 5. FUNDING NUMBERS C - F30602-97-C-0259

  1. 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...

  2. 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.

  3. 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.

  4. 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

  5. MARTe: A Multiplatform Real-Time Framework

    Science.gov (United States)

    Neto, André C.; Sartori, Filippo; Piccolo, Fabio; Vitelli, Riccardo; De Tommasi, Gianmaria; Zabeo, Luca; Barbalace, Antonio; Fernandes, Horacio; Valcarcel, Daniel F.; Batista, Antonio J. N.

    2010-04-01

    Development of real-time applications is usually associated with nonportable code targeted at specific real-time operating systems. The boundary between hardware drivers, system services, and user code is commonly not well defined, making the development in the target host significantly difficult. The Multithreaded Application Real-Time executor (MARTe) is a framework built over a multiplatform library that allows the execution of the same code in different operating systems. The framework provides the high-level interfaces with hardware, external configuration programs, and user interfaces, assuring at the same time hard real-time performances. End-users of the framework are required to define and implement algorithms inside a well-defined block of software, named Generic Application Module (GAM), that is executed by the real-time scheduler. Each GAM is reconfigurable with a set of predefined configuration meta-parameters and interchanges information using a set of data pipes that are provided as inputs and required as output. Using these connections, different GAMs can be chained either in series or parallel. GAMs can be developed and debugged in a non-real-time system and, only once the robustness of the code and correctness of the algorithm are verified, deployed to the real-time system. The software also supplies a large set of utilities that greatly ease the interaction and debugging of a running system. Among the most useful are a highly efficient real-time logger, HTTP introspection of real-time objects, and HTTP remote configuration. MARTe is currently being used to successfully drive the plasma vertical stabilization controller on the largest magnetic confinement fusion device in the world, with a control loop cycle of 50 ?s and a jitter under 1 ?s. In this particular project, MARTe is used with the Real-Time Application Interface (RTAI)/Linux operating system exploiting the new ?86 multicore processors technology.

  6. Detection of ROS1 Gene Rearrangement in Lung Adenocarcinoma: Comparison of IHC, FISH and Real-Time RT-PCR

    OpenAIRE

    Shan, Ling; Lian, Fang; Guo, Lei; Qiu, Tian; Ling, Yun; Ying, Jianming; Lin, Dongmei

    2015-01-01

    Aims To compare fluorescence in situ hybridization (FISH), immunohistochemistry (IHC) and quantitative real-time reverse transcription-PCR (qRT-PCR) assays for detection of ROS1 fusion in a large number of ROS1-positive lung adenocatcinoma (ADC) patients. Methods Using IHC analysis, sixty lung ADCs including 16 cases with ROS1 protein expression and 44 cases without ROS1 expression were selected for this study. The ROS1 fusion status was examined by FISH and qRT-PCR assay. Results Among 60 ca...

  7. Mapping of single-copy genes by TSA-FISH in the codling moth, Cydia pomonella.

    Science.gov (United States)

    Carabajal Paladino, Leonela Z; Nguyen, Petr; Síchová, Jindra; Marec, František

    2014-01-01

    We work on the development of transgenic sexing strains in the codling moth, Cydia pomonella (Tortricidae), which would enable to produce male-only progeny for the population control of this pest using sterile insect technique (SIT). To facilitate this research, we have developed a number of cytogenetic and molecular tools, including a physical map of the codling moth Z chromosome using BAC-FISH (fluorescence in situ hybridization with bacterial artificial chromosome probes). However, chromosomal localization of unique, single-copy sequences such as a transgene cassette by conventional FISH remains challenging. In this study, we adapted a FISH protocol with tyramide signal amplification (TSA-FISH) for detection of single-copy genes in Lepidoptera. We tested the protocol with probes prepared from partial sequences of Z-linked genes in the codling moth. Using a modified TSA-FISH protocol we successfully mapped a partial sequence of the Acetylcholinesterase 1 (Ace-1) gene to the Z chromosome and confirmed thus its Z-linkage. A subsequent combination of BAC-FISH with BAC probes containing anticipated neighbouring Z-linked genes and TSA-FISH with the Ace-1 probe allowed the integration of Ace-1 in the physical map of the codling moth Z chromosome. We also developed a two-colour TSA-FISH protocol which enabled us simultaneous localization of two Z-linked genes, Ace-1 and Notch, to the expected regions of the Z chromosome. We showed that TSA-FISH represents a reliable technique for physical mapping of genes on chromosomes of moths and butterflies. Our results suggest that this technique can be combined with BAC-FISH and in the future used for physical localization of transgene cassettes on chromosomes of transgenic lines in the codling moth or other lepidopteran species. Furthermore, the developed protocol for two-colour TSA-FISH might become a powerful tool for synteny mapping in non-model organisms.

  8. Research of real-time communication software

    Science.gov (United States)

    Li, Maotang; Guo, Jingbo; Liu, Yuzhong; Li, Jiahong

    2003-11-01

    Real-time communication has been playing an increasingly important role in our work, life and ocean monitor. With the rapid progress of computer and communication technique as well as the miniaturization of communication system, it is needed to develop the adaptable and reliable real-time communication software in the ocean monitor system. This paper involves the real-time communication software research based on the point-to-point satellite intercommunication system. The object-oriented design method is adopted, which can transmit and receive video data and audio data as well as engineering data by satellite channel. In the real-time communication software, some software modules are developed, which can realize the point-to-point satellite intercommunication in the ocean monitor system. There are three advantages for the real-time communication software. One is that the real-time communication software increases the reliability of the point-to-point satellite intercommunication system working. Second is that some optional parameters are intercalated, which greatly increases the flexibility of the system working. Third is that some hardware is substituted by the real-time communication software, which not only decrease the expense of the system and promotes the miniaturization of communication system, but also aggrandizes the agility of the system.

  9. Making real-time reactive systems reliable

    Science.gov (United States)

    Marzullo, Keith; Wood, Mark

    1990-01-01

    A reactive system is characterized by a control program that interacts with an environment (or controlled program). The control program monitors the environment and reacts to significant events by sending commands to the environment. This structure is quite general. Not only are most embedded real time systems reactive systems, but so are monitoring and debugging systems and distributed application management systems. Since reactive systems are usually long running and may control physical equipment, fault tolerance is vital. The research tries to understand the principal issues of fault tolerance in real time reactive systems and to build tools that allow a programmer to design reliable, real time reactive systems. In order to make real time reactive systems reliable, several issues must be addressed: (1) How can a control program be built to tolerate failures of sensors and actuators. To achieve this, a methodology was developed for transforming a control program that references physical value into one that tolerates sensors that can fail and can return inaccurate values; (2) How can the real time reactive system be built to tolerate failures of the control program. Towards this goal, whether the techniques presented can be extended to real time reactive systems is investigated; and (3) How can the environment be specified in a way that is useful for writing a control program. Towards this goal, whether a system with real time constraints can be expressed as an equivalent system without such constraints is also investigated.

  10. Real time sensor for therapeutic radiation delivery

    International Nuclear Information System (INIS)

    Bliss, M.; Craig, R.A.; Reeder, P.L.

    1998-01-01

    The invention is a real time sensor for therapeutic radiation. A probe is placed in or near the patient that senses in real time the dose at the location of the probe. The strength of the dose is determined by either an insertion or an exit probe. The location is determined by a series of vertical and horizontal sensing elements that gives the operator a real time read out dose location relative to placement of the patient. The increased accuracy prevents serious tissue damage to the patient by preventing overdose or delivery of a dose to a wrong location within the body. 14 figs

  11. Real time detecting system for turning force

    Energy Technology Data Exchange (ETDEWEB)

    Xiaobin, Yue [China Academy of Engineering Physics, Mianyang (China). Inst. of Machinery Manufacturing Technology

    2001-07-01

    How to get the real-time value of forces dropped on the tool in the course of processing by piezoelectric sensors is introduced. First, the analog signals of the cutting force were achieved by these sensors, amplified and transferred into digital signals by A/D transferring card. Then real-time software reads the information, put it into its own coordinate, drew the curve of forces, displayed it on the screen by the real time and saved it for the technicians to analyze the situation of the tool. So the cutting parameter can be optimized to improve surface quality of the pieces.

  12. 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.

  13. Coordinating Transit Transfers in Real Time

    Science.gov (United States)

    2016-05-06

    Transfers are a major source of travel time variability for transit passengers. Coordinating transfers between transit routes in real time can reduce passenger waiting times and travel time variability, but these benefits need to be contrasted with t...

  14. Real-Time Penetrating Particle Analyzer (PAN)

    Science.gov (United States)

    Wu, X.; Ambrosi, G.; Bertucci, B.

    2018-02-01

    The PAN can measure penetrating particles with great precision to study energetic particles, solar activities, and the origin and propagation of cosmic rays. The real-time monitoring of penetrating particles is crucial for deep space human travel.

  15. Frequent dual initiation of reverse transcription in murine leukemia virus-based vectors containing two primer-binding sites

    International Nuclear Information System (INIS)

    Voronin, Yegor A.; Pathak, Vinay K.

    2003-01-01

    Retroviruses package two copies of viral RNA into each virion. Although each RNA contains a primer-binding site for initiation of DNA synthesis, it is unknown whether reverse transcription is initiated on both RNAs. To determine whether a single virion is capable of initiating reverse transcription more than once, we constructed a murine leukemia virus-based vector containing a second primer-binding site (PBS) derived from spleen necrosis virus and inserted the green fluorescent protein gene (GFP) between the two PBSs. Initiation of reverse transcription at either PBS results in a provirus that expresses GFP. However, initiation at both PBSs can result in the deletion of GFP, which can be detected by flow cytometry and Southern blotting analysis. Approximately 22-29% of the proviruses formed deleted the GFP in a single replication cycle, indicating the minimum proportion of virions that initiated reverse transcription on both PBSs. These results show that a significant proportion of MLV-based vectors containing two PBSs have the capacity to initiate reverse transcription more than once

  16. 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 language...... 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....

  17. 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)

  18. Identification of valid reference genes for gene expression studies of human stomach cancer by reverse transcription-qPCR

    International Nuclear Information System (INIS)

    Rho, Hyun-Wook; Lee, Byoung-Chan; Choi, Eun-Seok; Choi, Il-Ju; Lee, Yeon-Su; Goh, Sung-Ho

    2010-01-01

    Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful method for the analysis of gene expression. Target gene expression levels are usually normalized to a consistently expressed reference gene also known as internal standard, in the same sample. However, much effort has not been expended thus far in the search for reference genes suitable for the study of stomach cancer using RT-qPCR, although selection of optimal reference genes is critical for interpretation of results. We assessed the suitability of six possible reference genes, beta-actin (ACTB), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyl transferase 1 (HPRT1), beta-2-microglobulin (B2M), ribosomal subunit L29 (RPL29) and 18S ribosomal RNA (18S rRNA) in 20 normal and tumor stomach tissue pairs of stomach cancer patients and 6 stomach cancer cell lines, by RT-qPCR. Employing expression stability analyses using NormFinder and geNorm algorithms we determined the order of performance of these reference genes and their variation values. This RT-qPCR study showed that there are statistically significant (p < 0.05) differences in the expression levels of HPRT1 and 18S rRNA in 'normal-' versus 'tumor stomach tissues'. The stability analyses by geNorm suggest B2M-GAPDH, as best reference gene combination for 'stomach cancer cell lines'; RPL29-HPRT1, for 'all stomach tissues'; and ACTB-18S rRNA, for 'all stomach cell lines and tissues'. NormFinder also identified B2M as the best reference gene for 'stomach cancer cell lines', RPL29-B2M for 'all stomach tissues', and 18S rRNA-ACTB for 'all stomach cell lines and tissues'. The comparisons of normalized expression of the target gene, GPNMB, showed different interpretation of target gene expression depend on best single reference gene or combination. This study validated RPL29 and RPL29-B2M as the best single reference

  19. Benefits of real-time gas management

    International Nuclear Information System (INIS)

    Nolty, R.; Dolezalek, D. Jr.

    1994-01-01

    In today's competitive gas gathering, processing, storage and transportation business environment, the requirements to do business are continually changing. These changes arise from government regulations such as the amendments to the Clean Air Act concerning the environment and FERC Order 636 concerning business practices. Other changes are due to advances in technology such as electronic flow measurement (EFM) and real-time communications capabilities within the gas industry. Gas gathering, processing, storage and transportation companies must be flexible in adapting to these changes to remain competitive. These dynamic requirements can be met with an open, real-time gas management computer information system. Such a system provides flexible services with a variety of software applications. Allocations, nominations management and gas dispatching are examples of applications that are provided on a real-time basis. By providing real-time services, the gas management system enables operations personnel to make timely adjustments within the current accounting period. Benefits realized from implementing a real-time gas management system include reduced unaccountable gas, reduced imbalance penalties, reduced regulatory violations, improved facility operations and better service to customers. These benefits give a company the competitive edge. This article discusses the applications provided, the benefits from implementing a real-time gas management system, and the definition of such a system

  20. Typing of Poultry Influenza Virus (H5 and H7 by Reverse Transcription- Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Cesare Bonacina

    2010-01-01

    Full Text Available The ability of the influenza Orthomixovirus to undergo to continually antigenically changes that can affect its pathogenicity and its diffusion, explains the growing seriousness of this disease and the recent epizoozies in various parts of the world. There have been 15 HA and 9 NA type A sub-types of the influenza virus identified all of which are present in birds. Until now the very virulent avian influenza viruses identified were all included to the H5 and H7 sub-types. We here show that is possible to identify the H5 and H7 sub-types with reverse transcription-polymerase chain reaction (RT-PCR by using a set of specific primers for each HA sub-type. The RT-PCR is a quick and sensitive method of identifying the HA sub-types of the influenza virus directly from homogenised organs.

  1. Application of anti-listerial bacteriocins: monitoring enterocin expression by multiplex relative reverse transcription-PCR.

    Science.gov (United States)

    Williams, D Ross; Chanos, Panagiotis

    2012-12-01

    Listeriosis is a deadly food-borne disease, and its incidence may be limited through the biotechnological exploitation of a number of anti-listerial biocontrol agents. The most widely used of these agents are bacteriocins and the Class II enterocins are characterized by their activity against Listeria. Enterocins are primarily produced by enterococci, particularly Enterococcus faecium and many strains have been described, often encoding multiple bacteriocins. The use of these strains in food will require that they are free of virulence functions and that they exhibit a high level expression of anti-listerial enterocins in fermentation conditions. Multiplex relative RT (reverse transcription)-PCR is a technique that is useful in the discovery of advantageous expression characteristics among enterocin-producing strains. It allows the levels of individual enterocin gene expression to be monitored and determination of how expression is altered under different growth conditions.

  2. Real-time PCR in virology.

    Science.gov (United States)

    Mackay, Ian M; Arden, Katherine E; Nitsche, Andreas

    2002-03-15

    The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of PCR product during real-time PCR. These are the DNA binding fluorophores, the 5' endonuclease, adjacent linear and hairpin oligoprobes and the self-fluorescing amplicons, which are described in detail. We also discuss factors that have restricted the development of multiplex real-time PCR as well as the role of real-time PCR in quantitating nucleic acids. Both amplification hardware and the fluorogenic detection chemistries have evolved rapidly as the understanding of real-time PCR has developed and this review aims to update the scientist on the current state of the art. We describe the background, advantages and limitations of real-time PCR and we review the literature as it applies to virus detection in the routine and research laboratory in order to focus on one of the many areas in which the application of real-time PCR has provided significant methodological benefits and improved patient outcomes. However, the technology discussed has been applied to other areas of microbiology as well as studies of gene expression and genetic disease.

  3. 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.

  4. Structure of the gene for human butyrylcholinesterase. Evidence for a single copy

    International Nuclear Information System (INIS)

    Arpagaus, M.; Kott, M.; Vatsis, K.P.; Bartels, C.F.; La Du, B.N.; Lockridge, O.

    1990-01-01

    The authors have isolated five genomic clones for human butyrylcholinesterase (BChE), using cDNA probes encoding the catalytic subunit of the hydrophilic tetramer. The BChE gene is at least 73 kb long and contains for exons. Exon 1 contains untranslated sequences and two potential translation initiation sites at codons -69 and -47. Exon 2 (1525 bp) contains 83% of the coding sequence for the mature protein, including the N-terminal and the active-site serine, and a third possible translation initiation site (likely functional), at codon -28. Exon 3 is 167 nucleotides long. Exon 4 (604 bp) codes for the C-terminus of the protein and the 3' untranslated region where two polyadenylation signals were identified. Intron 1 is 6.5 km long, and the minimal sizes of introns 2 and 3 are estimated to be 32 km each. Southern blot analysis of total human genomic DNA is in complete agreement with the gene structure established by restriction endonuclease mapping of the genomic clones: this strongly suggests that the BChE gene is present in a single copy

  5. Phylogeny of the cycads based on multiple single copy nuclear genes: congruence of concatenation and species tree inference methods

    Science.gov (United States)

    Despite a recent new classification, a stable tree of life for the cycads has been elusive, particularly regarding resolution of Bowenia, Stangeria and Dioon. In this study we apply five single copy nuclear genes (SCNGs) to the phylogeny of the order Cycadales. We specifically aim to evaluate seve...

  6. Testing of real-time-software

    International Nuclear Information System (INIS)

    Friesland, G.; Ovenhausen, H.

    1975-05-01

    The situation in the area of testing real-time-software is unsatisfactory. During the first phase of the project PROMOTE (prozessorientiertes Modul- und Gesamttestsystem) an analysis of the momentary situation took place, results of which are summarized in the following study about some user interviews and an analysis of relevant literature. 22 users (industry, software-houses, hardware-manufacturers, and institutes) have been interviewed. Discussions were held about reliability of real-time software with special interest to error avoidance, testing, and debugging. Main aims of the analysis of the literature were elaboration of standard terms, comparison of existing test methods and -systems, and the definition of boundaries to related areas. During the further steps of this project some means and techniques will be worked out to systematically test real-time software. (orig.) [de

  7. 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)

  8. 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

  9. Axial Tomography from Digitized Real Time Radiography

    Science.gov (United States)

    Zolnay, A. S.; McDonald, W. M.; Doupont, P. A.; McKinney, R. L.; Lee, M. M.

    1985-01-18

    Axial tomography from digitized real time radiographs provides a useful tool for industrial radiography and tomography. The components of this system are: x-ray source, image intensifier, video camera, video line extractor and digitizer, data storage and reconstruction computers. With this system it is possible to view a two dimensional x-ray image in real time at each angle of rotation and select the tomography plane of interest by choosing which video line to digitize. The digitization of a video line requires less than a second making data acquisition relatively short. Further improvements on this system are planned and initial results are reported.

  10. Replacing OSE with Real Time capable Linux

    OpenAIRE

    Boman, Simon; Rutgersson, Olof

    2009-01-01

    For many years OSE has been a common used operating system, with real time extensions enhancements, in embed-ded systems. But in the last decades, Linux has grown and became a competitor to common operating systems and, in recent years, even as an operating system with real time extensions. With this in mind, ÅF was interested in replacing the quite expensive OSE with some distribution of the open source based Linux on a PowerPC MPC8360. Therefore, our purpose with thesis is to implement Linu...

  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. 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

  13. Real-time ISEE data system

    Science.gov (United States)

    Tsurutani, B. T.; Baker, D. N.

    1979-01-01

    A real-time ISEE data system directed toward predicting geomagnetic substorms and storms is discussed. Such a system may allow up to 60+ minutes advance warning of magnetospheric substorms and up to 30 minute warnings of geomagnetic storms (and other disturbances) induced by high-speed streams and solar flares. The proposed system utilizes existing capabilities of several agencies (NASA, NOAA, USAF), and thereby minimizes costs. This same concept may be applicable to data from other spacecraft, and other NASA centers; thus, each individual experimenter can receive quick-look data in real time at his or her base institution.

  14. Mobile waste inspection real time radiography system

    International Nuclear Information System (INIS)

    Vigil, J.; Taggart, D.; Betts, S.; Rael, C.; Martinez, F.; Mendez, J.

    1995-01-01

    The 450-KeV Mobile Real Time Radiography System was designed and purchased to inspect containers of radioactive waste produced at Los Alamos National Laboratory (LANL). The Mobile Real Time Radiography System has the capability of inspecting waste containers of various sizes from 5-gal. buckets to standard waste boxes (SWB, dimensions 54.5 in. x 71 in. x 37 in.). The fact that this unit is mobile makes it an attractive alternative to the costly road closures associated with moving waste from the waste generator to storage or disposal facilities

  15. 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.

  16. Automated real-time software development

    Science.gov (United States)

    Jones, Denise R.; Walker, Carrie K.; Turkovich, John J.

    1993-01-01

    A Computer-Aided Software Engineering (CASE) system has been developed at the Charles Stark Draper Laboratory (CSDL) under the direction of the NASA Langley Research Center. The CSDL CASE tool provides an automated method of generating source code and hard copy documentation from functional application engineering specifications. The goal is to significantly reduce the cost of developing and maintaining real-time scientific and engineering software while increasing system reliability. This paper describes CSDL CASE and discusses demonstrations that used the tool to automatically generate real-time application code.

  17. Rapid diagnostic detection of plum pox virus in Prunus plants by isothermal AmplifyRP(®) using reverse transcription-recombinase polymerase amplification.

    Science.gov (United States)

    Zhang, Shulu; Ravelonandro, Michel; Russell, Paul; McOwen, Nathan; Briard, Pascal; Bohannon, Seven; Vrient, Albert

    2014-10-01

    Plum pox virus (PPV) causes the most destructive viral disease known as plum pox or Sharka disease in stone fruit trees. As an important regulated pathogen, detection of PPV is thus of critical importance to quarantine and eradication of the spreading disease. In this study, the innovative development of two AmplifyRP(®) tests is reported for a rapid isothermal detection of PPV using reverse transcription-recombinase polymerase amplification. In an AmplifyRP(®) test, all specific recombination and amplification reactions occur at a constant temperature without thermal cycling and the test results are either recorded in real-time with a portable fluorescence reader or displayed using a lateral flow strip contained inside an amplicon detection chamber. The major improvement of this assay is that the entire test from sample preparation to result can be completed in as little as 20min and can be performed easily both in laboratories and in the field. The results from this study demonstrated the ability of the AmplifyRP(®) technique to detect all nine PPV strains (An, C, CR, D, EA, M, Rec, T, or W). Among the economic benefits to pathogen surveys is the higher sensitivity of the AmplifyRP(®) to detect PPV when compared to the conventional ELISA and ImmunoStrip(®) assays. This is the first report describing the use of such an innovative technique to detect rapidly plant viruses affecting perennial crops. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Selection and evaluation of potential reference genes for gene expression analysis in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae using reverse-transcription quantitative PCR.

    Directory of Open Access Journals (Sweden)

    Miao Yuan

    Full Text Available The brown planthopper (BPH, Nilaparvata lugens (Hemiptera, Delphacidae, is one of the most important rice pests. Abundant genetic studies on BPH have been conducted using reverse-transcription quantitative real-time PCR (qRT-PCR. Using qRT-PCR, the expression levels of target genes are calculated on the basis of endogenous controls. These genes need to be appropriately selected by experimentally assessing whether they are stably expressed under different conditions. However, such studies on potential reference genes in N. lugens are lacking. In this paper, we presented a systematic exploration of eight candidate reference genes in N. lugens, namely, actin 1 (ACT, muscle actin (MACT, ribosomal protein S11 (RPS11, ribosomal protein S15e (RPS15, alpha 2-tubulin (TUB, elongation factor 1 delta (EF, 18S ribosomal RNA (18S, and arginine kinase (AK and used four alternative methods (BestKeeper, geNorm, NormFinder, and the delta Ct method to evaluate the suitability of these genes as endogenous controls. We examined their expression levels among different experimental factors (developmental stage, body part, geographic population, temperature variation, pesticide exposure, diet change, and starvation following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments guidelines. Based on the results of RefFinder, which integrates four currently available major software programs to compare and rank the tested candidate reference genes, RPS15, RPS11, and TUB were found to be the most suitable reference genes in different developmental stages, body parts, and geographic populations, respectively. RPS15 was the most suitable gene under different temperature and diet conditions, while RPS11 was the most suitable gene under different pesticide exposure and starvation conditions. This work sheds light on establishing a standardized qRT-PCR procedure in N. lugens, and serves as a starting point for screening for reference genes for

  19. Single-copy nuclear genes resolve the phylogeny of the holometabolous insects

    Directory of Open Access Journals (Sweden)

    Bertone Matthew A

    2009-06-01

    Full Text Available Abstract Background Evolutionary relationships among the 11 extant orders of insects that undergo complete metamorphosis, called Holometabola, remain either unresolved or contentious, but are extremely important as a context for accurate comparative biology of insect model organisms. The most phylogenetically enigmatic holometabolan insects are Strepsiptera or twisted wing parasites, whose evolutionary relationship to any other insect order is unconfirmed. They have been controversially proposed as the closest relatives of the flies, based on rDNA, and a possible homeotic transformation in the common ancestor of both groups that would make the reduced forewings of Strepsiptera homologous to the reduced hindwings of Diptera. Here we present evidence from nucleotide sequences of six single-copy nuclear protein coding genes used to reconstruct phylogenetic relationships and estimate evolutionary divergence times for all holometabolan orders. Results Our results strongly support Hymenoptera as the earliest branching holometabolan lineage, the monophyly of the extant orders, including the fleas, and traditionally recognized groupings of Neuropteroidea and Mecopterida. Most significantly, we find strong support for a close relationship between Coleoptera (beetles and Strepsiptera, a previously proposed, but analytically controversial relationship. Exploratory analyses reveal that this relationship cannot be explained by long-branch attraction or other systematic biases. Bayesian divergence times analysis, with reference to specific fossil constraints, places the origin of Holometabola in the Carboniferous (355 Ma, a date significantly older than previous paleontological and morphological phylogenetic reconstructions. The origin and diversification of most extant insect orders began in the Triassic, but flourished in the Jurassic, with multiple adaptive radiations producing the astounding diversity of insect species for which these groups are so well

  20. 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

  1. 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.

  2. 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.

  3. 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...

  4. 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...

  5. 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...

  6. 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…

  7. 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...

  8. 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

  9. 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.

  10. Model Checking Real-Time Systems

    DEFF Research Database (Denmark)

    Bouyer, Patricia; Fahrenberg, Uli; Larsen, Kim Guldstrand

    2018-01-01

    This chapter surveys timed automata as a formalism for model checking real-time systems. We begin with introducing the model, as an extension of finite-state automata with real-valued variables for measuring time. We then present the main model-checking results in this framework, and give a hint...

  11. Real-time systems design and analysis

    CERN Document Server

    Laplante, Phillip A

    2004-01-01

    "Real-Time Systems Design and Analysis, Third Edition is essential for students and practicing software engineers who want improved designs, faster computation, and ultimate cost savings. Chapters discuss hardware considerations and software requirements, software systems design, the software production process, performance estimation and optimization, and engineering considerations."--Jacket.

  12. Testing Real-Time Systems Using UPPAAL

    DEFF Research Database (Denmark)

    Hessel, Anders; Larsen, Kim Guldstrand; Mikucionis, Marius

    2008-01-01

    This chapter presents principles and techniques for model-based black-box conformance testing of real-time systems using the Uppaal model-checking tool-suite. The basis for testing is given as a network of concurrent timed automata specified by the test engineer. Relativized input...

  13. Temporal logics and real time expert systems

    NARCIS (Netherlands)

    Blom, J.A.

    1996-01-01

    This paper introduces temporal logics. Due to the eternal compromise between expressive adequacy and reasoning efficiency that must decided upon in any application, full (first order logic or modal logic based) temporal logics are frequently not suitable. This is especially true in real time expert

  14. Ray Tracing for Real-time Games

    NARCIS (Netherlands)

    Bikker, J.

    2012-01-01

    This thesis describes efficient rendering algorithms based on ray tracing, and the application of these algorithms to real-time games. Compared to rasterizationbased approaches, rendering based on ray tracing allows elegant and correct simulation of important global effects, such as shadows,

  15. 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

  16. 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...

  17. Composing Synchronisation and Real-Time Constraints

    NARCIS (Netherlands)

    Bergmans, Lodewijk; Aksit, Mehmet

    There have been a number of publications illustrating the successes of object-oriented techniques in creating highly reusable software systems. Several concurrent languages have been proposed for specifying reusable synchronization specifications. Recently, a number of real-time object-oriented

  18. In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells

    DEFF Research Database (Denmark)

    Lange, Marianne; Tolker-Nielsen, Tim; Molin, Søren

    2000-01-01

    An in situ reverse transcription-PCR protocol for detecting specific mRNA in Methanosarcina mazei S-6 is described. This method allowed us to detect heat shock-induced increases in the intracellular levels of the transcript of the universal stress gene dnaK. The cell walls of paraformaldehyde...

  19. Concepts of real time and semi-real time material control

    International Nuclear Information System (INIS)

    Lovett, J.E.

    1975-01-01

    After a brief consideration of the traditional material balance accounting on an MBA basis, this paper explores the basic concepts of real time and semi-real time material control, together with some of the major problems to be solved. Three types of short-term material control are discussed: storage, batch processing, and continuous processing. (DLC)

  20. Reverse transcription and polymerase chain reaction: principles and applications in dentistry.

    Science.gov (United States)

    Santos, Carlos Ferreira Dos; Sakai, Vivien Thiemy; Machado, Maria Aparecida de Andrade Moreira; Schippers, Daniela Nicole; Greene, Andrew Seth

    2004-03-01

    Various molecular biology techniques have become available in the last few years. One of the most revolutionary of these techniques regarding nucleic acid analysis is the polymerase chain reaction (PCR), which was first described in 1985. This method relies on the exponential amplification of specific DNA fragments, resulting in millions of copies that can serve as templates for different kinds of analyses. PCR can be preceded by a reverse transcription (RT) reaction in order to produce cDNA from RNA (RT-PCR). RT-PCR provides the possibility to assess gene transcription in cells or tissues. PCR and RT-PCR techniques have been instrumental in dental research, and show potential to be used for diagnosis as well as for treatment and prevention of many diseases (dental caries, periodontal disease, endodontic infections and oral cancer). Compared to other traditional methodologies, PCR and RT-PCR show many advantages including high specificity, sensitivity, and speed. Since PCR and RT-PCR are relatively new techniques and are not available to most students and professionals involved with dentistry, the aim of this work is to present the details of these techniques as well as dental literature reports in which they were used.

  1. Identification of a methylated oligoribonucleotide as a potent inhibitor of HIV-1 reverse transcription complex.

    Science.gov (United States)

    Grigorov, Boyan; Bocquin, Anne; Gabus, Caroline; Avilov, Sergey; Mély, Yves; Agopian, Audrey; Divita, Gilles; Gottikh, Marina; Witvrouw, Myriam; Darlix, Jean-Luc

    2011-07-01

    Upon HIV-1 infection of a target cell, the viral reverse transcriptase (RT) copies the genomic RNA to synthesize the viral DNA. The genomic RNA is within the incoming HIV-1 core where it is coated by molecules of nucleocapsid (NC) protein that chaperones the reverse transcription process. Indeed, the RT chaperoning properties of NC extend from the initiation of cDNA synthesis to completion of the viral DNA. New and effective drugs against HIV-1 continue to be required, which prompted us to search for compounds aimed at inhibiting NC protein. Here, we report that the NC chaperoning activity is extensively inhibited in vitro by small methylated oligoribonucleotides (mODN). These mODNs were delivered intracellularly using a cell-penetrating-peptide and found to impede HIV-1 replication in primary human cells at nanomolar concentrations. Extensive analysis showed that viral cDNA synthesis was severely impaired by mODNs. Partially resistant viruses with mutations in NC and RT emerged after months of passaging in cell culture. A HIV-1 molecular clone (NL4.3) bearing these mutations was found to replicate at high concentrations of mODN, albeit with a reduced fitness. Small, methylated ODNs such as mODN-11 appear to be a new type of highly potent inhibitor of HIV-1.

  2. Detection of canine cytokine gene expression by reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Pinelli, E; van der Kaaij, S Y; Slappendel, R; Fragio, C; Ruitenberg, E J; Bernadina, W; Rutten, V P

    1999-08-02

    Further characterization of the canine immune system will greatly benefit from the availability of tools to detect canine cytokines. Our interest concerns the study on the role of cytokines in canine visceral leishmaniasis. For this purpose, we have designed specific primers using previously published sequences for the detection of canine IL-2, IFN-gamma and IL10 mRNA by reverse transcription-polymerase chain reaction (RT-PCR). For IL-4, we have cloned and sequenced this cytokine gene, and developed canine-specific primers. To control for sample-to-sample variation in the quantity of mRNA and variation in the RT and PCR reactions, the mRNA levels of glyceraldehyde-3-phosphate dehydrogenase (G3PDH), a housekeeping gene, were determined in parallel. Primers to amplify G3PDH were designed from consensus sequences obtained from the Genbank database. The mRNA levels of the cytokines mentioned here were detected from ConA-stimulated peripheral mononuclear cells derived from Leishmania-infected dogs. A different pattern of cytokine production among infected animals was found.

  3. Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers

    International Nuclear Information System (INIS)

    Ryu, Jin Sook; Moon, Dae Hyuk; Cheon, Jun Hong; Chung, Yoon Young; Park, Hung Dong; Chung, Young Hwa; Lee, Young Sang

    1994-01-01

    This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patients serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was detected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows, 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79% ) of the same group with conventional method 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possibility of false positivity and false negativity.

  4. Detection of Tilapia Lake Virus in Clinical Samples by Culturing and Nested Reverse Transcription-PCR.

    Science.gov (United States)

    Kembou Tsofack, Japhette Esther; Zamostiano, Rachel; Watted, Salsabeel; Berkowitz, Asaf; Rosenbluth, Ezra; Mishra, Nischay; Briese, Thomas; Lipkin, W Ian; Kabuusu, Richard M; Ferguson, Hugh; Del Pozo, Jorge; Eldar, Avi; Bacharach, Eran

    2017-03-01

    Tilapia are an important group of farmed fish that serve as a significant protein source worldwide. In recent years, substantial mortality of wild tilapia has been observed in the Sea of Galilee and in commercial ponds in Israel and Ecuador. We have identified the etiological agent of these mass die-offs as a novel orthomyxo-like virus and named it tilapia lake virus (TiLV). Here, we provide the conditions for efficient isolation, culturing, and quantification of the virus, including the use of susceptible fish cell lines. Moreover, we describe a sensitive nested reverse transcription-PCR (RT-PCR) assay allowing the rapid detection of TiLV in fish organs. This assay revealed, for the first time to our knowledge, the presence of TiLV in diseased Colombian tilapia, indicating a wider distribution of this emerging pathogen and stressing the risk that TiLV poses for the global tilapia industry. Overall, the described procedures should provide the tilapia aquaculture industry with important tools for the detection and containment of this pathogen. Copyright © 2017 American Society for Microbiology.

  5. Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Jin Sook; Moon, Dae Hyuk; Cheon, Jun Hong; Chung, Yoon Young; Park, Hung Dong; Chung, Young Hwa; Lee, Young Sang [Asan Medical Center, University of Ulsan, Seoul (Korea, Republic of)

    1994-07-15

    This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patients serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was detected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows, 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79% ) of the same group with conventional method 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possibility of false positivity and false negativity.

  6. Detection of Tilapia Lake Virus in Clinical Samples by Culturing and Nested Reverse Transcription-PCR

    Science.gov (United States)

    Kembou Tsofack, Japhette Esther; Zamostiano, Rachel; Watted, Salsabeel; Berkowitz, Asaf; Rosenbluth, Ezra; Mishra, Nischay; Briese, Thomas; Lipkin, W. Ian; Kabuusu, Richard M.; Ferguson, Hugh; del Pozo, Jorge

    2016-01-01

    ABSTRACT Tilapia are an important group of farmed fish that serve as a significant protein source worldwide. In recent years, substantial mortality of wild tilapia has been observed in the Sea of Galilee and in commercial ponds in Israel and Ecuador. We have identified the etiological agent of these mass die-offs as a novel orthomyxo-like virus and named it tilapia lake virus (TiLV). Here, we provide the conditions for efficient isolation, culturing, and quantification of the virus, including the use of susceptible fish cell lines. Moreover, we describe a sensitive nested reverse transcription-PCR (RT-PCR) assay allowing the rapid detection of TiLV in fish organs. This assay revealed, for the first time to our knowledge, the presence of TiLV in diseased Colombian tilapia, indicating a wider distribution of this emerging pathogen and stressing the risk that TiLV poses for the global tilapia industry. Overall, the described procedures should provide the tilapia aquaculture industry with important tools for the detection and containment of this pathogen. PMID:27974544

  7. 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.

  8. Real-time ISEE data system

    International Nuclear Information System (INIS)

    Tsurutani, B.T.; Baker, D.N.

    1979-01-01

    Prediction of geomagnetic substorms and storms would be of great scientific and commercial interest. A real-time ISEE data system directed toward this purpose is discussed in detail. Such a system may allow up to 60+ minutes advance warning of magnetospheric substorms and up to 30 minute warnings of geomagnetic storms (and other disturbances) induced by high-speed streams and solar flares. The proposed system utilizes existing capabilities of several agencies (NASA, NOAA, USAF), and thereby minimizes costs. This same concept may be applicable to data from other spacecraft, and other NASA centers; thus, each individual experimenter can receive quick-look data in real time at his or her base institution. 6 figures, 1 table

  9. Real time animation of space plasma phenomena

    International Nuclear Information System (INIS)

    Jordan, K.F.; Greenstadt, E.W.

    1987-01-01

    In pursuit of real time animation of computer simulated space plasma phenomena, the code was rewritten for the Massively Parallel Processor (MPP). The program creates a dynamic representation of the global bowshock which is based on actual spacecraft data and designed for three dimensional graphic output. This output consists of time slice sequences which make up the frames of the animation. With the MPP, 16384, 512 or 4 frames can be calculated simultaneously depending upon which characteristic is being computed. The run time was greatly reduced which promotes the rapid sequence of images and makes real time animation a foreseeable goal. The addition of more complex phenomenology in the constructed computer images is now possible and work proceeds to generate these images

  10. Real Time Radiation Monitoring Using Nanotechnology

    Science.gov (United States)

    Li, Jing (Inventor); Hanratty, James J. (Inventor); Wilkins, Richard T. (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.

  11. 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.

  12. 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.

  13. Real time processor for array speckle interferometry

    International Nuclear Information System (INIS)

    Chin, G.; Florez, J.; Borelli, R.; Fong, W.; Miko, J.; Trujillo, C.

    1989-01-01

    With the construction of several new large aperture telescopes and the development of large format array detectors in the near IR, the ability to obtain diffraction limited seeing via IR array speckle interferometry offers a powerful tool. We are constructing a real-time processor to acquire image frames, perform array flat-fielding, execute a 64 x 64 element 2D complex FFT, and to average the power spectrum all within the 25 msec coherence time for speckles at near IR wavelength. The processor is a compact unit controlled by a PC with real time display and data storage capability. It provides the ability to optimize observations and obtain results on the telescope rather than waiting several weeks before the data can be analyzed and viewed with off-line methods

  14. Real time processor for array speckle interferometry

    Science.gov (United States)

    Chin, Gordon; Florez, Jose; Borelli, Renan; Fong, Wai; Miko, Joseph; Trujillo, Carlos

    1989-02-01

    The authors are constructing a real-time processor to acquire image frames, perform array flat-fielding, execute a 64 x 64 element two-dimensional complex FFT (fast Fourier transform) and average the power spectrum, all within the 25 ms coherence time for speckles at near-IR (infrared) wavelength. The processor will be a compact unit controlled by a PC with real-time display and data storage capability. This will provide the ability to optimize observations and obtain results on the telescope rather than waiting several weeks before the data can be analyzed and viewed with offline methods. The image acquisition and processing, design criteria, and processor architecture are described.

  15. Real Time Structured Light and Applications

    DEFF Research Database (Denmark)

    Wilm, Jakob

    Structured light scanning is a versatile method for 3D shape acquisition. While much faster than most competing measurement techniques, most high-end structured light scans still take in the order of seconds to complete. Low-cost sensors such as Microsoft Kinect and time of flight cameras have made......, 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...

  16. 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)

  17. Real-time imaging of quantum entanglement.

    Science.gov (United States)

    Fickler, Robert; Krenn, Mario; Lapkiewicz, Radek; Ramelow, Sven; Zeilinger, Anton

    2013-01-01

    Quantum Entanglement is widely regarded as one of the most prominent features of quantum mechanics and quantum information science. Although, photonic entanglement is routinely studied in many experiments nowadays, its signature has been out of the grasp for real-time imaging. Here we show that modern technology, namely triggered intensified charge coupled device (ICCD) cameras are fast and sensitive enough to image in real-time the effect of the measurement of one photon on its entangled partner. To quantitatively verify the non-classicality of the measurements we determine the detected photon number and error margin from the registered intensity image within a certain region. Additionally, the use of the ICCD camera allows us to demonstrate the high flexibility of the setup in creating any desired spatial-mode entanglement, which suggests as well that visual imaging in quantum optics not only provides a better intuitive understanding of entanglement but will improve applications of quantum science.

  18. 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)

  19. Real time simulator for material testing reactor

    Energy Technology Data Exchange (ETDEWEB)

    Takemoto, Noriyuki; Imaizumi, Tomomi; Izumo, Hironobu; Hori, Naohiko; Suzuki, Masahide [Japan Atomic Energy Agency, Oarai Research and Development Center, Oarai, Ibaraki (Japan); Ishitsuka, Tatsuo; Tamura, Kazuo [ITOCHU Techno-Solutions Corp., Tokyo (Japan)

    2012-03-15

    Japan Atomic Energy Agency (JAEA) is now developing a real time simulator for a material testing reactor based on Japan Materials Testing Reactor (JMTR). The simulator treats reactor core system, primary and secondary cooling system, electricity system and irradiation facility systems. Possible simulations are normal reactor operation, unusual transient operation and accidental operation. The developed simulator also contains tool to revise/add facility in it for the future development. (author)

  20. Probabilistic real-time contingency ranking method

    International Nuclear Information System (INIS)

    Mijuskovic, N.A.; Stojnic, D.

    2000-01-01

    This paper describes a real-time contingency method based on a probabilistic index-expected energy not supplied. This way it is possible to take into account the stochastic nature of the electric power system equipment outages. This approach enables more comprehensive ranking of contingencies and it is possible to form reliability cost values that can form the basis for hourly spot price calculations. The electric power system of Serbia is used as an example for the method proposed. (author)

  1. Advanced real time radioscopy and computed tomography

    International Nuclear Information System (INIS)

    Sauerwein, Ch.; Nuding, W.; Grimm, R.; Wiacker, H.

    1996-01-01

    The paper describes three x-ray inspection systems. One radioscopic system is designed for the inspection of castings. The next integrates a radioscopic and a tomographic mode. The radioscopy has a high resolution camera and real time image processor. Radiation sources are a 450 kV industrial and a 200 kV microfocus tube. The third system is a tomographic system with 30 scintillation detectors for the inspection of nuclear waste containers. (author)

  2. Real time computer controlled weld skate

    Science.gov (United States)

    Wall, W. A., Jr.

    1977-01-01

    A real time, adaptive control, automatic welding system was developed. This system utilizes the general case geometrical relationships between a weldment and a weld skate to precisely maintain constant weld speed and torch angle along a contoured workplace. The system is compatible with the gas tungsten arc weld process or can be adapted to other weld processes. Heli-arc cutting and machine tool routing operations are possible applications.

  3. 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...

  4. Telepositional portable real time radiation monitoring system

    International Nuclear Information System (INIS)

    Talpalariu, Jeni; Matei, Corina; Popescu, Oana

    2010-01-01

    Technology development for complex portable networks is on going to meet the area dosimetry challenge, improving the basic design using new telepositional GPS satellite methods and GSM terrestrial civil radio transmission networks. The system and devices proposed overcome the limitations of fixed and portable dosimeters, providing wireless real time radiations data and geospatial information's means, using many portable dosimeter stations and a mobile dosimeter computerised central console. (authors)

  5. Real time simulator for material testing reactor

    International Nuclear Information System (INIS)

    Takemoto, Noriyuki; Imaizumi, Tomomi; Izumo, Hironobu; Hori, Naohiko; Suzuki, Masahide; Ishitsuka, Tatsuo; Tamura, Kazuo

    2012-01-01

    Japan Atomic Energy Agency (JAEA) is now developing a real time simulator for a material testing reactor based on Japan Materials Testing Reactor (JMTR). The simulator treats reactor core system, primary and secondary cooling system, electricity system and irradiation facility systems. Possible simulations are normal reactor operation, unusual transient operation and accidental operation. The developed simulator also contains tool to revise/add facility in it for the future development. (author)

  6. Boundary Correct Real-Time Soft Shadows

    DEFF Research Database (Denmark)

    Jacobsen, Bjarke; Christensen, Niels Jørgen; Larsen, Bent Dalgaard

    2004-01-01

    This paper describes a method to determine correct shadow boundaries from an area light source using umbra and penumbra volumes. The light source is approximated by a circular disk as this gives a fast way to extrude the volumes. The method also gives a crude estimate of the visibility of the are...... for implementation on most programmable hardware. Though some crude approximations are used in the visibility function, the method can be used to produce soft shadows with correct boundaries in real time....

  7. Real-time Astrometry Using Phase Congruency

    Science.gov (United States)

    Lambert, A.; Polo, M.; Tang, Y.

    Phase congruency is a computer vision technique that proves to perform well for determining the tracks of optical objects (Flewelling, AMOS 2014). We report on a real-time implementation of this using an FPGA and CMOS Image Sensor, with on-sky data. The lightweight instrument can provide tracking update signals to the mount of the telescope, as well as determine abnormal objects in the scene.

  8. Real-time multiple image manipulations

    International Nuclear Information System (INIS)

    Arenson, J.S.; Shalev, S.; Legris, J.; Goertzen, Y.

    1984-01-01

    There are many situations in which it is desired to manipulate two or more images under real-time operator control. The authors have investigated a number of such cases in order to determine their value and applicability in clinical medicine and laboratory research. Several examples are presented in detail. The DICOM-8 video image computer system was used due to its capability of storing two 512 x 512 x 8 bit images and operating on them, and/or an incoming video frame, with any of a number of real time operations including addition, subtraction, inversion, averaging, logical AND, NAND, OR, NOR, NOT, XOR and XNOR, as well as combinations of these. Some applications involve manipulations of or among the stored images. In others, a stored image is used as a mask or template for positioning or adjusting a second image to be grabbed via a video camera. The accuracy of radiotherapy treatment is verified by comparing port films with the original radiographic planning film, which is previously digitized and stored. Moving the port film on the light box while viewing the real-time subtraction image allows for adjustments of zoom, translation and rotation, together with contrast and edge enhancement

  9. Real time ultrasonography in obstructive jaundice

    International Nuclear Information System (INIS)

    Cho, Kyung Sik; Kim, Ho Kyun; Sung, Nak Kwan; Kim, Soon Yong

    1982-01-01

    Ultrasonography is a predominantly accurate, relatively simple unique diagnostic method of obstructive jaundice. The ultrasonographic findings of obstructive jaundice are dilated intra- and extrahepatic duct with intraluminal hyper reflective echo or mass in and/ or around the bile duct. The superiority of high resolution real time ultrasonography for the diagnosis of obstructive jaundice is bases on the easy detectability of extra- and intrahepatic bile ducts by its multiple sectional images in a short time, the flexibility of probe and small crystal size. Author evaluated real time sonographic findings 46 obstructive jaundice patients confirmed by surgery or radiographical examinations. The results were: 1. Diameter of extrahepatic duct in obstructive jaundice were varied from normal to 4.0 Cm, mostly 8 to 10 mm in diameter (26%). Degree of dilatation of biliary duct appeared more prominent in cancer patients than other causes of obstruction. 2. The site of obstruction was detected in 85% (39/46) and its common site was common bile duct in 63% (29/46). 3. The diagnostic accuracy of choledocholithiasis and cancer was 82% (22/27) and 44% (4/9), respectively. Diagnostic accuracy of the real time ultrasonography in obstructive jaundice was over all 75% (34/46)

  10. Real-time video quality monitoring

    Science.gov (United States)

    Liu, Tao; Narvekar, Niranjan; Wang, Beibei; Ding, Ran; Zou, Dekun; Cash, Glenn; Bhagavathy, Sitaram; Bloom, Jeffrey

    2011-12-01

    The ITU-T Recommendation G.1070 is a standardized opinion model for video telephony applications that uses video bitrate, frame rate, and packet-loss rate to measure the video quality. However, this model was original designed as an offline quality planning tool. It cannot be directly used for quality monitoring since the above three input parameters are not readily available within a network or at the decoder. And there is a great room for the performance improvement of this quality metric. In this article, we present a real-time video quality monitoring solution based on this Recommendation. We first propose a scheme to efficiently estimate the three parameters from video bitstreams, so that it can be used as a real-time video quality monitoring tool. Furthermore, an enhanced algorithm based on the G.1070 model that provides more accurate quality prediction is proposed. Finally, to use this metric in real-world applications, we present an example emerging application of real-time quality measurement to the management of transmitted videos, especially those delivered to mobile devices.

  11. Real-time earthquake data feasible

    Science.gov (United States)

    Bush, Susan

    Scientists agree that early warning devices and monitoring of both Hurricane Hugo and the Mt. Pinatubo volcanic eruption saved thousands of lives. What would it take to develop this sort of early warning and monitoring system for earthquake activity?Not all that much, claims a panel assigned to study the feasibility, costs, and technology needed to establish a real-time earthquake monitoring (RTEM) system. The panel, drafted by the National Academy of Science's Committee on Seismology, has presented its findings in Real-Time Earthquake Monitoring. The recently released report states that “present technology is entirely capable of recording and processing data so as to provide real-time information, enabling people to mitigate somewhat the earthquake disaster.” RTEM systems would consist of two parts—an early warning system that would give a few seconds warning before severe shaking, and immediate postquake information within minutes of the quake that would give actual measurements of the magnitude. At this time, however, this type of warning system has not been addressed at the national level for the United States and is not included in the National Earthquake Hazard Reduction Program, according to the report.

  12. The Raptor Real-Time Processing Architecture

    Science.gov (United States)

    Galassi, M.; Starr, D.; Wozniak, P.; Brozdin, K.

    The primary goal of Raptor is ambitious: to identify interesting optical transients from very wide field of view telescopes in real time, and then to quickly point the higher resolution Raptor ``fovea'' cameras and spectrometer to the location of the optical transient. The most interesting of Raptor's many applications is the real-time search for orphan optical counterparts of Gamma Ray Bursts. The sequence of steps (data acquisition, basic calibration, source extraction, astrometry, relative photometry, the smarts of transient identification and elimination of false positives, telescope pointing feedback, etc.) is implemented with a ``component'' approach. All basic elements of the pipeline functionality have been written from scratch or adapted (as in the case of SExtractor for source extraction) to form a consistent modern API operating on memory resident images and source lists. The result is a pipeline which meets our real-time requirements and which can easily operate as a monolithic or distributed processing system. Finally, the Raptor architecture is entirely based on free software (sometimes referred to as ``open source'' software). In this paper we also discuss the interplay between various free software technologies in this type of astronomical problem.

  13. Tuning Linux to meet real time requirements

    Science.gov (United States)

    Herbel, Richard S.; Le, Dang N.

    2007-04-01

    There is a desire to use Linux in military systems. Customers are requesting contractors to use open source to the maximal possible extent in contracts. Linux is probably the best operating system of choice to meet this need. It is widely used. It is free. It is royalty free, and, best of all, it is completely open source. However, there is a problem. Linux was not originally built to be a real time operating system. There are many places where interrupts can and will be blocked for an indeterminate amount of time. There have been several attempts to bridge this gap. One of them is from RTLinux, which attempts to build a microkernel underneath Linux. The microkernel will handle all interrupts and then pass it up to the Linux operating system. This does insure good interrupt latency; however, it is not free [1]. Another is RTAI, which provides a similar typed interface; however, the PowerPC platform, which is used widely in real time embedded community, was stated as "recovering" [2]. Thus this is not suited for military usage. This paper provides a method for tuning a standard Linux kernel so it can meet the real time requirement of an embedded system.

  14. Real-time RT-PCR, a necessary tool to support the diagnosis and surveillance of rotavirus in Mexico.

    Science.gov (United States)

    De La Cruz Hernández, Sergio Isaac; Anaya Molina, Yazmin; Gómez Santiago, Fabián; Terán Vega, Heidi Lizbeth; Monroy Leyva, Elda; Méndez Pérez, Héctor; García Lozano, Herlinda

    2018-04-01

    Rotavirus produces diarrhea in children under 5 years old. Most of those conventional methods such as polyacrylamide gel electrophoresis (PAGE) and reverse transcription-polymerase chain reaction (RT-PCR) have been used for rotavirus detection. However, these techniques need a multi-step process to get the results. In comparison with conventional methods, the real-time RT-PCR is a highly sensitive method, which allows getting the results in only one day. In this study a real-time RT-PCR assay was tested using a panel of 440 samples from patients with acute gastroenteritis, and characterized by PAGE and RT-PCR. The results show that the real-time RT-PCR detected rotavirus from 73% of rotavirus-negative samples analyzed by PAGE and RT-PCR; thus, the percentage of rotavirus-positive samples increased to 81%. The results indicate that this real-time RT-PCR should be part of a routine analysis, and as a support of the diagnosis of rotavirus in Mexico. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. A Simple Reverse Transcription-Polymerase Chain Reaction for Dengue Type 2 Virus Identification

    Directory of Open Access Journals (Sweden)

    Luiz Tadeu M Figueiredo

    1997-05-01

    Full Text Available We show here a simplified reverse transcription-polymerase chain reaction (RT-PCR for identification of dengue type 2 virus. Three dengue type 2 virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD, as a negative control, were used in this study. C6/36 cells were infected with the virus, and tissue culture fluids were collected after 7 days of infection period. The RT-PCR, a combination of RT and PCR done after a single addition of reagents in a single reaction vessel was carried out following a digestion of virus with 1% Nonidet P-40. The 50ml assay reaction mixture included 50 pmol of a dengue type 2 specific primer pair amplifying a 210 base pair sequence of the envelope protein gene, 0.1 mM of the four deoxynucleoside triphosphates, 7.5U of reverse transcriptase, and 1U of thermostable Taq DNA polymerase. The reagent mixture was incubated for 15 min at 37oC for RT followed by a variable amount of cycles of two-step PCR amplification (92oC for 60 sec, 53oC for 60 sec with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized with UV light after gel incubation in ethidium bromide solution. DNA bands were observed after 25 and 30 cycles of PCR. Virus amount as low as 102.8 TCID50/ml was detected by RT-PCR. Specific DNA amplification was observed with the three dengue type 2 strains. This assay has advantages compared to other RT-PCRs: it avoids laborious extraction of virus RNA; the combination of RT and PCR reduces assay time, facilitates the performance and reduces risk of contamination; the two-step PCR cycle produces a clear DNA amplification, saves assay time and simplifies the technique

  16. Reverse Transcription Errors and RNA-DNA Differences at Short Tandem Repeats.

    Science.gov (United States)

    Fungtammasan, Arkarachai; Tomaszkiewicz, Marta; Campos-Sánchez, Rebeca; Eckert, Kristin A; DeGiorgio, Michael; Makova, Kateryna D

    2016-10-01

    Transcript variation has important implications for organismal function in health and disease. Most transcriptome studies focus on assessing variation in gene expression levels and isoform representation. Variation at the level of transcript sequence is caused by RNA editing and transcription errors, and leads to nongenetically encoded transcript variants, or RNA-DNA differences (RDDs). Such variation has been understudied, in part because its detection is obscured by reverse transcription (RT) and sequencing errors. It has only been evaluated for intertranscript base substitution differences. Here, we investigated transcript sequence variation for short tandem repeats (STRs). We developed the first maximum-likelihood estimator (MLE) to infer RT error and RDD rates, taking next generation sequencing error rates into account. Using the MLE, we empirically evaluated RT error and RDD rates for STRs in a large-scale DNA and RNA replicated sequencing experiment conducted in a primate species. The RT error rates increased exponentially with STR length and were biased toward expansions. The RDD rates were approximately 1 order of magnitude lower than the RT error rates. The RT error rates estimated with the MLE from a primate data set were concordant with those estimated with an independent method, barcoded RNA sequencing, from a Caenorhabditis elegans data set. Our results have important implications for medical genomics, as STR allelic variation is associated with >40 diseases. STR nonallelic transcript variation can also contribute to disease phenotype. The MLE and empirical rates presented here can be used to evaluate the probability of disease-associated transcripts arising due to RDD. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  17. Comparison of multiplex RT-PCR and real-time HybProbe assay for serotyping of dengue virus using reference strains and clinical samples from India

    Directory of Open Access Journals (Sweden)

    Anita Chakravarti

    2016-01-01

    Full Text Available Background: Dengue virus serotyping is crucial from clinical management and epidemiological point of view. Aims: To compare efficacy of two molecular detection and typing methods, namely, multiplex reverse transcription polymerase chain reaction (RT-PCR and real-time Hybprobe assay using a panel of known dilution of four reference Dengue virus strains and a panel of sera collected from clinically suspected dengue patients. Settings: This study was conducted at a tertiary-care teaching hospital in Delhi, India. Materials and Methods: Dengue serotype specific virus strains were used as prototypes for serotyping assays. Viral load was quantified by quantitative real time reverse transcription polymerase chain reaction (qRT-PCR. Acute phase serum samples were collected from 79 patients with clinically suspected Dengue fever on their first day of presentation during September-October 2012. Viral RNA from serum and cell culture supernatant was extracted. Reverse transcription was carried out. Quantitative detection of DENV RNA from reference strain culture supernatants and each of the 79 patient samples by real-time PCR was performed using light cycler Taqman master mix kit. Serotyping was done by multiplex RT-PCR assay and Hybprobe assay. Results: The multiplex RT-PCR assay, though found to be 100% specific, couldn't serotype either patient or reference strains with viral load less than 1000 RNA copies/ml. The Hybprobe assay was found to have 100% specificity and had a lower limit of serotype detection of merely 3.54 RNA copies/ml. Conclusions: HybProbe assay has an important role especially in situations where serotyping is to be performed in clinical samples with low viral load.

  18. Rapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification.

    Science.gov (United States)

    Zong, Xiaojuan; Wang, Wenwen; Wei, Hairong; Wang, Jiawei; Chen, Xin; Xu, Li; Zhu, Dongzi; Tan, Yue; Liu, Qingzhong

    2014-11-01

    Prunus necrotic ringspot virus (PNRSV) has seriously reduced the yield of Prunus species worldwide. In this study, a highly efficient and specific two-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect PNRSV. Total RNA was extracted from sweet cherry leaf samples using a commercial kit based on a magnetic nanoparticle technique. Transcripts were used as the templates for the assay. The results of this assay can be detected using agarose gel electrophoresis or by assessing in-tube fluorescence after adding SYBR Green I. The assay is highly specific for PNRSV, and it is more sensitive than reverse-transcription polymerase chain reaction (RT-PCR). Restriction enzyme digestion verified further the reliability of this RT-LAMP assay. To our knowledge, this is the first report of the application of RT-LAMP to PNRSV detection in Prunus species. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Exploring Earthquakes in Real-Time

    Science.gov (United States)

    Bravo, T. K.; Kafka, A. L.; Coleman, B.; Taber, J. J.

    2013-12-01

    Earthquakes capture the attention of students and inspire them to explore the Earth. Adding the ability to view and explore recordings of significant and newsworthy earthquakes in real-time makes the subject even more compelling. To address this opportunity, the Incorporated Research Institutions for Seismology (IRIS), in collaboration with Moravian College, developed ';jAmaSeis', a cross-platform application that enables students to access real-time earthquake waveform data. Students can watch as the seismic waves are recorded on their computer, and can be among the first to analyze the data from an earthquake. jAmaSeis facilitates student centered investigations of seismological concepts using either a low-cost educational seismograph or streamed data from other educational seismographs or from any seismic station that sends data to the IRIS Data Management System. After an earthquake, students can analyze the seismograms to determine characteristics of earthquakes such as time of occurrence, distance from the epicenter to the station, magnitude, and location. The software has been designed to provide graphical clues to guide students in the analysis and assist in their interpretations. Since jAmaSeis can simultaneously record up to three stations from anywhere on the planet, there are numerous opportunities for student driven investigations. For example, students can explore differences in the seismograms from different distances from an earthquake and compare waveforms from different azimuthal directions. Students can simultaneously monitor seismicity at a tectonic plate boundary and in the middle of the plate regardless of their school location. This can help students discover for themselves the ideas underlying seismic wave propagation, regional earthquake hazards, magnitude-frequency relationships, and the details of plate tectonics. The real-time nature of the data keeps the investigations dynamic, and offers students countless opportunities to explore.

  20. Simultaneous real-time data collection methods

    Science.gov (United States)

    Klincsek, Thomas

    1992-01-01

    This paper describes the development of electronic test equipment which executes, supervises, and reports on various tests. This validation process uses computers to analyze test results and report conclusions. The test equipment consists of an electronics component and the data collection and reporting unit. The PC software, display screens, and real-time data-base are described. Pass-fail procedures and data replay are discussed. The OS2 operating system and Presentation Manager user interface system were used to create a highly interactive automated system. The system outputs are hardcopy printouts and MS DOS format files which may be used as input for other PC programs.

  1. Temporal logics and real time expert systems.

    Science.gov (United States)

    Blom, J A

    1996-10-01

    This paper introduces temporal logics. Due to the eternal compromise between expressive adequacy and reasoning efficiency that must decided upon in any application, full (first order logic or modal logic based) temporal logics are frequently not suitable. This is especially true in real time expert systems, where a fixed (and usually small) response time must be guaranteed. One such expert system, Fagan's VM, is reviewed, and a delineation is given of how to formally describe and reason with time in medical protocols. It is shown that Petri net theory is a useful tool to check the correctness of formalised protocols.

  2. CUDA-based real time surgery simulation.

    Science.gov (United States)

    Liu, Youquan; De, Suvranu

    2008-01-01

    In this paper we present a general software platform that enables real time surgery simulation on the newly available compute unified device architecture (CUDA)from NVIDIA. CUDA-enabled GPUs harness the power of 128 processors which allow data parallel computations. Compared to the previous GPGPU, it is significantly more flexible with a C language interface. We report implementation of both collision detection and consequent deformation computation algorithms. Our test results indicate that the CUDA enables a twenty times speedup for collision detection and about fifteen times speedup for deformation computation on an Intel Core 2 Quad 2.66 GHz machine with GeForce 8800 GTX.

  3. Real-time PCR in virology

    OpenAIRE

    Mackay, Ian M.; Arden, Katherine E.; Nitsche, Andreas

    2002-01-01

    The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of P...

  4. Systems Analyze Water Quality in Real Time

    Science.gov (United States)

    2010-01-01

    A water analyzer developed under Small Business Innovation Research (SBIR) contracts with Kennedy Space Center now monitors treatment processes at water and wastewater facilities around the world. Originally designed to provide real-time detection of nutrient levels in hydroponic solutions for growing plants in space, the ChemScan analyzer, produced by ASA Analytics Inc., of Waukesha, Wisconsin, utilizes spectrometry and chemometric algorithms to automatically analyze multiple parameters in the water treatment process with little need for maintenance, calibration, or operator intervention. The company has experienced a compound annual growth rate of 40 percent over its 15-year history as a direct result of the technology's success.

  5. Real-Time Thevenin Impedance Computation

    DEFF Research Database (Denmark)

    Sommer, Stefan Horst; Jóhannsson, Hjörtur

    2013-01-01

    operating state, and strict time constraints are difficult to adhere to as the complexity of the grid increases. Several suggested approaches for real-time stability assessment require Thevenin impedances to be determined for the observed system conditions. By combining matrix factorization, graph reduction......, and parallelization, we develop an algorithm for computing Thevenin impedances an order of magnitude faster than previous approaches. We test the factor-and-solve algorithm with data from several power grids of varying complexity, and we show how the algorithm allows realtime stability assessment of complex power...

  6. Real-time modeling of heat distributions

    Science.gov (United States)

    Hamann, Hendrik F.; Li, Hongfei; Yarlanki, Srinivas

    2018-01-02

    Techniques for real-time modeling temperature distributions based on streaming sensor data are provided. In one aspect, a method for creating a three-dimensional temperature distribution model for a room having a floor and a ceiling is provided. The method includes the following steps. A ceiling temperature distribution in the room is determined. A floor temperature distribution in the room is determined. An interpolation between the ceiling temperature distribution and the floor temperature distribution is used to obtain the three-dimensional temperature distribution model for the room.

  7. Linear Regression Based Real-Time Filtering

    Directory of Open Access Journals (Sweden)

    Misel Batmend

    2013-01-01

    Full Text Available This paper introduces real time filtering method based on linear least squares fitted line. Method can be used in case that a filtered signal is linear. This constraint narrows a band of potential applications. Advantage over Kalman filter is that it is computationally less expensive. The paper further deals with application of introduced method on filtering data used to evaluate a position of engraved material with respect to engraving machine. The filter was implemented to the CNC engraving machine control system. Experiments showing its performance are included.

  8. A real-time Global Warming Index.

    Science.gov (United States)

    Haustein, K; Allen, M R; Forster, P M; Otto, F E L; Mitchell, D M; Matthews, H D; Frame, D J

    2017-11-13

    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 human-induced warming is compatible with a range of other more sophisticated methods to estimate the human contribution to observed global temperature change.

  9. General purpose computers in real time

    International Nuclear Information System (INIS)

    Biel, J.R.

    1989-01-01

    I see three main trends in the use of general purpose computers in real time. The first is more processing power. The second is the use of higher speed interconnects between computers (allowing more data to be delivered to the processors). The third is the use of larger programs running in the computers. Although there is still work that needs to be done, I believe that all indications are that the online need for general purpose computers should be available for the SCC and LHC machines. 2 figs

  10. Detection of Brazilian hantavirus by reverse transcription polymerase chain reaction amplification of N gene in patients with hantavirus cardiopulmonary syndrome

    OpenAIRE

    Marcos Lázaro Moreli; Ricardo Luiz Moro de Sousa; Luiz Tadeu Moraes Figueiredo

    2004-01-01

    We report a nested reverse transcription-polymerase chain reaction (RT-PCR) assay for hantavirus using primers selected to match high homology regions of hantavirus genomes detected from the whole blood of hantavirus cardiopulmonary syndrome (HCPS) patients from Brazil, also including the N gene nucleotide sequence of Araraquara virus. Hantavirus genomes were detected in eight out of nine blood samples from the HCPS patients by RT-PCR (88.9% positivity) and in all 9 blood samples (100% positi...

  11. Detection of Enterovirus 71 gene from clinical specimens by reverse-transcription loop-mediated isothermal amplification

    OpenAIRE

    D Wang; X Wang; Y Geng; C An

    2014-01-01

    Purpose : The objective of this study was to develop a sensitive, specific and rapid approach to diagnose hand foot and mouth disease (HFMD) for an early treatment by using loop-mediated isothermal amplification (LAMP) technique. Materials and Methods : A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for detecting EV71 virus was developed, the specificity and sensitivity of RT-LAMP was tested, and the clinical specimens was assayed by the RT-LAMP comparing with conven...

  12. Selection of suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using quantitative real-time polymerase chain reaction

    DEFF Research Database (Denmark)

    Zornhagen, K. W.; Kristensen, A. T.; Hansen, Anders Elias

    2015-01-01

    Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a sensitive technique for quantifying gene expression. Stably expressed reference genes are necessary for normalization of RT-qPCR data. Only a few articles have been published on reference genes in canine tumours....... The objective of this study was to demonstrate how to identify suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using RT-qPCR. Primer pairs for 17 potential reference genes were designed and tested in archival tumour biopsies from six dogs. The geNorm algorithm...

  13. Development of mRNA-based body fluid identification using reverse transcription loop-mediated isothermal amplification.

    Science.gov (United States)

    Satoh, Tetsuya; Kouroki, Seiya; Ogawa, Keita; Tanaka, Yorika; Matsumura, Kazutoshi; Iwase, Susumu

    2018-04-25

    Identifying body fluids from forensic samples can provide valuable evidence for criminal investigations. Messenger RNA (mRNA)-based body fluid identification was recently developed, and highly sensitive parallel identification using reverse transcription polymerase chain reaction (RT-PCR) has been described. In this study, we developed reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a simple, rapid assay for identifying three common forensic body fluids, namely blood, semen, and saliva, and evaluated its specificity and sensitivity. Hemoglobin beta (HBB), transglutaminase 4 (TGM4), and statherin (STATH) were selected as marker genes for blood, semen, and saliva, respectively. RT-LAMP could be performed in a single step including both reverse transcription and DNA amplification under an isothermal condition within 60 min, and detection could be conveniently performed via visual fluorescence. Marker-specific amplification was performed in each assay, and no cross-reaction was observed among five representative forensically relevant body fluids. The detection limits of the assays were 0.3 nL, 30 nL, and 0.3 μL for blood, semen, and saliva, respectively, and their sensitivities were comparable with those of RT-PCR. Furthermore, RT-LAMP assays were applicable to forensic casework samples. It is considered that RT-LAMP is useful for body fluid identification.

  14. Software Design Methods for Real-Time Systems

    Science.gov (United States)

    1989-12-01

    This module describes the concepts and methods used in the software design of real time systems . It outlines the characteristics of real time systems , describes...the role of software design in real time system development, surveys and compares some software design methods for real - time systems , and

  15. Real-time inextensible surgical thread simulation.

    Science.gov (United States)

    Xu, Lang; Liu, Qian

    2018-03-27

    This paper discusses a real-time simulation method of inextensible surgical thread based on the Cosserat rod theory using position-based dynamics (PBD). The method realizes stable twining and knotting of surgical thread while including inextensibility, bending, twisting and coupling effects. The Cosserat rod theory is used to model the nonlinear elastic behavior of surgical thread. The surgical thread model is solved with PBD to achieve a real-time, extremely stable simulation. Due to the one-dimensional linear structure of surgical thread, the direct solution of the distance constraint based on tridiagonal matrix algorithm is used to enhance stretching resistance in every constraint projection iteration. In addition, continuous collision detection and collision response guarantee a large time step and high performance. Furthermore, friction is integrated into the constraint projection process to stabilize the twining of multiple threads and complex contact situations. Through comparisons with existing methods, the surgical thread maintains constant length under large deformation after applying the direct distance constraint in our method. The twining and knotting of multiple threads correspond to stable solutions to contact and friction forces. A surgical suture scene is also modeled to demonstrate the practicality and simplicity of our method. Our method achieves stable and fast simulation of inextensible surgical thread. Benefiting from the unified particle framework, the rigid body, elastic rod, and soft body can be simultaneously simulated. The method is appropriate for applications in virtual surgery that require multiple dynamic bodies.

  16. Real-time applications of neural nets

    International Nuclear Information System (INIS)

    Spencer, J.E.

    1989-05-01

    Producing, accelerating and colliding very high power, low emittance beams for long periods is a formidable problem in real-time control. As energy has grown exponentially in time so has the complexity of the machines and their control systems. Similar growth rates have occurred in many areas, e.g., improved integrated circuits have been paid for with comparable increases in complexity. However, in this case, reliability, capability and cost have improved due to reduced size, high production and increased integration which allow various kinds of feedback. In contrast, most large complex systems (LCS) are perceived to lack such possibilities because only one copy is made. Neural nets, as a metaphor for LCS, suggest ways to circumvent such limitations. It is argued that they are logically equivalent to multi-loop feedback/forward control of faulty systems. While complimentary to AI, they mesh nicely with characteristics desired for real-time systems. Such issues are considered, examples given and possibilities discussed. 21 refs., 6 figs

  17. A real-time radiation mapping system

    International Nuclear Information System (INIS)

    Scoggins, W.A.; VanEtten, D.M.

    1988-01-01

    A prototype of a real-time radiation mapping system, Ranger, was developed to respond to an accident involving the release of plutonium for the Department of Energy's Accident Response Group. In 1987 Ranger demonstrated that it can provide an efficient method of monitoring large areas of land for radioactive contamination. With the experience gained from the operation of the prototype, the external computer and software are being upgraded in order to obtain a fully operational system. The new system uses the prototype's commercially available line-of-sight microwave system for determining position and the same radiation detection instruments. The data obtained from the radiation detection instrument(s) are linked back to the external computer along with the relative position of the measurement through the ranging system. The data are displayed on a gridded map as colored circles and permanently stored in real-time. The different colors represent different contamination levels. Contours can be drawn using the permanently stored data. 4 figs

  18. Real time water chemistry monitoring and diagnostics

    International Nuclear Information System (INIS)

    Gaudreau, T.M.; Choi, S.S.

    2002-01-01

    EPRI has produced a real time water chemistry monitoring and diagnostic system. This system is called SMART ChemWorks and is based on the EPRI ChemWorks codes. System models, chemistry parameter relationships and diagnostic approaches from these codes are integrated with real time data collection, an intelligence engine and Internet technologies to allow for automated analysis of system chemistry. Significant data management capabilities are also included which allow the user to evaluate data and create automated reporting. Additional features have been added to the system in recent years including tracking and evaluation of primary chemistry as well as the calculation and tracking of primary to secondary leakage in PWRs. This system performs virtual sensing, identifies normal and upset conditions, and evaluates the consistency of on-line monitor and grab sample readings. The system also makes use of virtual fingerprinting to identify the cause of any chemistry upsets. This technology employs plant-specific data and models to determine the chemical state of the steam cycle. (authors)

  19. Interfacing real-time information with OILMAP

    International Nuclear Information System (INIS)

    Howlett, E.; Jayko, K.; Spaulding, M.

    1993-01-01

    OILMAP is a state-of-the-art, microcomputer-based oil spill response system applicable to oil spill contingency planning and real-time response for any location in the world. OILMAP has a graphic user interface and was designed in a modular framework so that different spill models could be incorporated into the system, as well as a suite of sophisticated data management tools, without increasing the complexity of the user interface. The basic OILMAP configuration contains a surface trajectory model intended for rapid, first-order estimates of spill movement. A variety of additional models are available within the OILMAP shell to address issues such as weathering, cleanup activities, and probabilities of oiling. A simplified geographic information system (GIS) allows display and manipulation of point, line, and area data geographically referenced to the spill domain. The GIS can import raster data so that images collected by satellite and aerial photography may be displayed. Several new capabilities have been implemented for OILMAP that allow real-time data to be integrated. These features include linking with the OILTRACKER free-floating buoys via a global positioning system, linking of hydrodynamic data from the Ocean Data and Information Network, the Harvard ocean forecasting system, and SeaSonde radar, and the capability of importing spill observations from any remotely sensed data. A further link between OILMAP's GIS and spill models has been developed which allows model predictions to be corrected to observed oil locations while the model runs. 13 refs., 6 figs

  20. Real-time applications of neural nets

    Energy Technology Data Exchange (ETDEWEB)

    Spencer, J.E.

    1989-05-01

    Producing, accelerating and colliding very high power, low emittance beams for long periods is a formidable problem in real-time control. As energy has grown exponentially in time so has the complexity of the machines and their control systems. Similar growth rates have occurred in many areas, e.g., improved integrated circuits have been paid for with comparable increases in complexity. However, in this case, reliability, capability and cost have improved due to reduced size, high production and increased integration which allow various kinds of feedback. In contrast, most large complex systems (LCS) are perceived to lack such possibilities because only one copy is made. Neural nets, as a metaphor for LCS, suggest ways to circumvent such limitations. It is argued that they are logically equivalent to multi-loop feedback/forward control of faulty systems. While complimentary to AI, they mesh nicely with characteristics desired for real-time systems. Such issues are considered, examples given and possibilities discussed. 21 refs., 6 figs.

  1. Real-time applications of neural nets

    International Nuclear Information System (INIS)

    Spencer, J.E.

    1989-01-01

    Producing, accelerating and colliding very high power, low emittance beams for long periods is a formidable problem in real-time control. As energy has grown exponentially in time so has the complexity of the machines and their control systems. Similar growth rates have occurred in many areas e.g. improved integrated circuits have been paid for with comparable increases in complexity. However, in this case, reliability, capability and cost have improved due to reduced size, high production and increased integration which allow various kinds of feedback. In contrast, most large complex systems (LCS) are perceived to lack such possibilities because only one copy is made. Neural nets, as a metaphor for LCS, suggest ways to circumvent such limitations. It is argued that they are logically equivalent to multi-loop feedback/forward control of faulty systems. While complimentary to AI, they mesh nicely with characteristics desired for real-time systems. In this paper, such issues are considered, examples given and possibilities discussed

  2. Detection of mRNA by reverse transcription PCR as an indicator of viability in Phytophthora ramorum

    Science.gov (United States)

    Antonio Chimento; Santa Olga Cacciola; Matteo Garbelotto

    2008-01-01

    Real-Time PCR technologies offer increasing opportunities to detect and study phytopathogenic fungi. They combine the sensitivity of conventional PCR with the generation of a specific fluorescent signal providing both real-time analysis of the reaction kinetics and quantification of specific DNA targets. Before the development of Real-Time PCR and...

  3. Memory controllers for real-time embedded systems predictable and composable real-time systems

    CERN Document Server

    Akesson, Benny

    2012-01-01

      Verification of real-time requirements in systems-on-chip becomes more complex as more applications are integrated. Predictable and composable systems can manage the increasing complexity using formal verification and simulation.  This book explains the concepts of predictability and composability and shows how to apply them to the design and analysis of a memory controller, which is a key component in any real-time system. This book is generally intended for readers interested in Systems-on-Chips with real-time applications.   It is especially well-suited for readers looking to use SDRAM memories in systems with hard or firm real-time requirements. There is a strong focus on real-time concepts, such as predictability and composability, as well as a brief discussion about memory controller architectures for high-performance computing. Readers will learn step-by-step how to go from an unpredictable SDRAM memory, offering highly variable bandwidth and latency, to a predictable and composable shared memory...

  4. Evaluation of Reference Genes for Reverse Transcription Quantitative PCR Studies of Physiological Responses in the Ghost Moth, Thitarodes armoricanus (Lepidoptera, Hepialidae.

    Directory of Open Access Journals (Sweden)

    Guiqing Liu

    Full Text Available Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR is the sensitive method to quantify the expression levels of target genes on the basis of endogenous control. An appropriate reference gene set for normalization is essential for reliable results. The ghost moth, Thitarodes armoricanus, a host species of a medicinal fungus, Ophiocordyceps sinensis, is an economically important member of the Lepidoptera. Recent studies have focused on the mechanism of adaptation of this species to its high-altitude environment and host immune response to O. sinensis infection and RT-qPCR is commonly used in these studies to decipher the genetic basis of physiological functions. However, a thorough assessment of candidate reference genes in the genus Thitarodes is lacking. Here, the expression levels of eight candidate reference genes (ACT, EF, EIF4A, GAPDH, G6PDH, RPL13A, TUB and 18S in T. armoricanus at different developmental stages and in different body parts of the seventh instar larvae were analyzed, along with larvae kept under low temperatures, larvae exposed to two fungal infections and larvae fed different diets. Three established software programs-Bestkeeper, geNorm and NormFinder-were employed to calculate variation among the treatments. The results revealed that the best-suited reference genes differed across the treatments, with EF, EIF4A and GAPDH found to be the best suited for the different developmental stages and larvae body parts; EF, EIF4A and RPL13A found to be the best suited for low-temperature challenge; and EF, EIF4A and TUB found to be the best suited for the fungal infections and dietary treatments. This study thus further contributes to the establishment of an accurate method for normalizing RT-qPCR results for T. armoricanus and serves as a reference for gene expression studies of related insect species.

  5. A field based detection method for Rose rosette virus using isothermal probe-based Reverse transcription-recombinase polymerase amplification assay.

    Science.gov (United States)

    Babu, Binoy; Washburn, Brian K; Ertek, Tülin Sarigül; Miller, Steven H; Riddle, Charles B; Knox, Gary W; Ochoa-Corona, Francisco M; Olson, Jennifer; Katırcıoğlu, Yakup Zekai; Paret, Mathews L

    2017-09-01

    Rose rosette disease, caused by Rose rosette virus (RRV; genus Emaravirus) is a major threat to the rose industry in the U.S. The only strategy currently available for disease management is early detection and eradication of the infected plants, thereby limiting its potential spread. Current RT-PCR based diagnostic methods for RRV are time consuming and are inconsistent in detecting the virus from symptomatic plants. Real-time RT-qPCR assay is highly sensitive for detection of RRV, but it is expensive and requires well-equipped laboratories. Both the RT-PCR and RT-qPCR cannot be used in a field-based testing for RRV. Hence a novel probe based, isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA) assay, using primer/probe designed based on the nucleocapsid gene of the RRV has been developed. The assay is highly specific and did not give a positive reaction to other viruses infecting roses belonging to both inclusive and exclusive genus. Dilution assays using the in vitro transcript showed that the primer/probe set is highly sensitive, with a detection limit of 1 fg/μl. In addition, a rapid technique for the extraction of viral RNA (rose varieties, collected from different states in the U.S. The entire process, including the extraction can be completed in 25min, with less sophisticated equipments. The developed assay can be used with high efficiency in large scale field testing for rapid detection of RRV in commercial nurseries and landscapes. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Reverse-transcriptional gene expression of anammox and ammonia-oxidizing archaea and bacteria in soybean and rice paddy soils of Northeast China.

    Science.gov (United States)

    Wang, Jing; Dong, Hailiang; Wang, Weidong; Gu, Ji-Dong

    2014-03-01

    The relative gene expression of hydrazine oxidoreductase encoding gene (hzo) for anaerobic ammonium oxidizing bacteria (anammox) and ammonia monooxygenase encoding gene (amoA) for both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in Sanjiang Plain soybean and rice paddy soils of Northeast China was investigated by using real-time reverse-transcriptional quantitative PCR. Metabolically active populations of anammox, AOA, and AOB in rice paddy soils were evident by the presence and successful quantification of hzo mRNA and amoA mRNA genes. The expression ratio of amoA gene for both AOA and AOB varied between soybean soils and different rice paddy soils while the expression of hzo gene for anammox was detectable only in rice paddy soils by showing a diverse relative expression ratio in each soil sample. Gene expression of both archaeal and bacterial amoA genes in rice paddy soils differed among the three sampling depths, but that of hzo was not. Both archaeal and bacterial amoA genes showed an increase trend of expression level with continuation of rice paddy cultivation, but the low expression ratio of hzo gene indicated a relatively small contribution of anammox in overall removal of inorganic nitrogen through N2 even under anoxic and high nitrogen input in agriculture. Bacterial amoA gene from two soybean fields and three rice paddy fields were also analyzed for community composition by denaturing gradient gel electrophoresis fingerprint. Community shift was observed between soybean and paddy fields and within each of them. The consistent occurrence of three bands 5, 6, and 7 in all samples showed their high adaptability for both arid cultivation and continuous rice paddy cultivation. Our data suggest that AOA and AOB are playing a more important role in nitrogen transformation in agricultural soils in oxic or anoxic environment and anammox bacteria may also contribute but in a less extent to N transformation in these agricultural soils

  7. Development of a multiplex amplification refractory mutation system reverse transcription polymerase chain reaction assay for the differential diagnosis of Feline leukemia virus vaccine and wild strains.

    Science.gov (United States)

    Ho, Chia-Fang; Chan, Kun-Wei; Yang, Wei-Cheng; Chiang, Yu-Chung; Chung, Yang-Tsung; Kuo, James; Wang, Chi-Young

    2014-07-01

    A multiplex amplification refractory mutation system reverse transcription polymerase chain reaction (ARMS RT-PCR) was developed for the differential diagnosis of Feline leukemia virus (FeLV) vaccine and wild-type strains based on a point mutation between the vaccine strain (S) and the wild-type strain (T) located in the p27 gene. This system was further upgraded to obtain a real-time ARMS RT-PCR (ARMS qRT-PCR) with a high-resolution melt analysis (HRMA) platform. The genotyping of various strains of FeLV was determined by comparing the HRMA curves with the defined wild-type FeLV (strain TW1), and the results were expressed as a percentage confidence. The detection limits of ARMS RT-PCR and ARMS qRT-PCR combined with HRMA were 100 and 1 copies of transcribed FeLV RNA per 0.5 ml of sample, respectively. No false-positive results were obtained with 6 unrelated pathogens and 1 feline cell line. Twelve FeLV Taiwan strains were correctly identified using ARMS qRT-PCR combined with HRMA. The genotypes of the strains matched the defined FeLV wild-type strain genotype with at least 91.17% confidence. A higher degree of sequence polymorphism was found throughout the p27 gene compared with the long terminal repeat region. In conclusion, the current study describes the phylogenetic relationship of the FeLV Taiwan strains and demonstrates that the developed ARMS RT-PCR assay is able to be used to detect the replication of a vaccine strain that has not been properly inactivated, thus acting as a safety check for the quality of FeLV vaccines.

  8. Field-Deployable Reverse Transcription-Insulated Isothermal PCR (RT-iiPCR) Assay for Rapid and Sensitive Detection of Foot-and-Mouth Disease Virus.

    Science.gov (United States)

    Ambagala, A; Fisher, M; Goolia, M; Nfon, C; Furukawa-Stoffer, T; Ortega Polo, R; Lung, O

    2017-10-01

    Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals, which can decimate the livestock industry and economy of countries previously free of this disease. Rapid detection of foot-and-mouth disease virus (FMDV) is critical to containing an FMD outbreak. Availability of a rapid, highly sensitive and specific, yet simple and field-deployable assay would support local decision-making during an FMDV outbreak. Here we report validation of a novel reverse transcription-insulated isothermal PCR (RT-iiPCR) assay that can be performed on a commercially available, compact and portable POCKIT ™ analyser that automatically analyses data and displays '+' or '-' results. The FMDV RT-iiPCR assay targets the 3D region of the FMDV genome and was capable of detecting 9 copies of in vitro-transcribed RNA standard with 95% confidence. It accurately identified 63 FMDV strains belonging to all seven serotypes and showed no cross-reactivity with viruses causing similar clinical diseases in cloven-hoofed animals. The assay was able to identify FMDV RNA in multiple sample types including oral, nasal and lesion swabs, epithelial tissue suspensions, vesicular and oral fluid samples, even before the appearance of clinical signs. Clinical sensitivity of the assay was comparable or slightly higher than the laboratory-based real-time RT-PCR assay in use. The assay was able to detect FMDV RNA in vesicular fluid samples without nucleic acid extraction. For RNA extraction from more complex sample types, a commercially available taco ™ mini transportable magnetic bead-based, automated extraction system was used. This assay provides a potentially useful field-deployable diagnostic tool for rapid detection of FMDV in an outbreak in FMD-free countries or for routine diagnostics in endemic countries with less structured laboratory systems. © 2016 Her Majesty the Queen in Right of Canada.

  9. Reverse transcription loop-mediated isothermal amplification assays for rapid identification of eastern and western strains of bluetongue virus in India.

    Science.gov (United States)

    Maan, S; Maan, N S; Batra, K; Kumar, A; Gupta, A; Rao, Panduranga P; Hemadri, Divakar; Reddy, Yella Narasimha; Guimera, M; Belaganahalli, M N; Mertens, P P C

    2016-08-01

    Bluetongue virus (BTV) infects all ruminants, including cattle, goats and camelids, causing bluetongue disease (BT) that is often severe in naïve deer and sheep. Reverse-transcription-loop-mediated-isothermal-amplification (RT-LAMP) assays were developed to detect eastern or western topotype of BTV strains circulating in India. Each assay uses four primers recognizing six distinct sequences of BTV genome-segment 1 (Seg-1). The eastern (e)RT-LAMP and western (w)RT-LAMP assay detected BTV RNA in all positive isolates that were tested (n=52, including Indian BTV-1, -2, -3, -5, -9, -10, -16, -21 -23, and -24 strains) with high specificity and efficiency. The analytical sensitivity of the RT-LAMP assays is comparable to real-time RT-PCR, but higher than conventional RT-PCR. The accelerated eRT-LAMP and wRT-LAMP assays generated detectable levels of amplified DNA, down to 0.216 fg of BTV RNA template or 108 fg of BTV RNA template within 60-90min respectively. The assays gave negative results with RNA from foot-and-mouth-disease virus (FMDV), peste des petits ruminants virus (PPRV), or DNA from Capripox viruses and Orf virus (n=10), all of which can cause clinical signs similar to BT. Both RT-LAMP assays did not show any cross-reaction among themselves. The assays are rapid, easy to perform, could be adapted as a 'penside' test making them suitable for 'front-line' diagnosis, helping to identify and contain field outbreaks of BTV. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. A novel method of multiple nucleic acid detection: Real-time RT-PCR coupled with probe-melting curve analysis.

    Science.gov (United States)

    Han, Yang; Hou, Shao-Yang; Ji, Shang-Zhi; Cheng, Juan; Zhang, Meng-Yue; He, Li-Juan; Ye, Xiang-Zhong; Li, Yi-Min; Zhang, Yi-Xuan

    2017-11-15

    A novel method, real-time reverse transcription PCR (real-time RT-PCR) coupled with probe-melting curve analysis, has been established to detect two kinds of samples within one fluorescence channel. Besides a conventional TaqMan probe, this method employs another specially designed melting-probe with a 5' terminus modification which meets the same label with the same fluorescent group. By using an asymmetric PCR method, the melting-probe is able to detect an extra sample in the melting stage effectively while it almost has little influence on the amplification detection. Thus, this method allows the availability of united employment of both amplification stage and melting stage for detecting samples in one reaction. The further demonstration by simultaneous detection of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) in one channel as a model system is presented in this essay. The sensitivity of detection by real-time RT-PCR coupled with probe-melting analysis was proved to be equal to that detected by conventional real-time RT-PCR. Because real-time RT-PCR coupled with probe-melting analysis can double the detection throughputs within one fluorescence channel, it is expected to be a good solution for the problem of low-throughput in current real-time PCR. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

    Directory of Open Access Journals (Sweden)

    I. Karthika Lakshmi

    2018-04-01

    Full Text Available Aim: The present study was designed to standardize real-time polymerase chain reaction (PCR for detecting the bluetongue virus from blood samples of sheep collected during outbreaks of bluetongue disease in the year 2014 in Andhra Pradesh and Telangana states of India. Materials and Methods: A 10-fold serial dilution of Plasmid PUC59 with bluetongue virus (BTV NS3 insert was used to plot the standard curve. BHK-21 and KC cells were used for in vitro propagation of virus BTV-9 at a TCID50/ml of 105 ml and RNA was isolated by the Trizol method. Both reverse transcription -PCR and real-time PCR using TaqMan probe were carried out with RNA extracted from virus-spiked culture medium and blood to compare the sensitivity by means of finding out the limit of detection (LoD. The results were verified by inoculating the detected and undetected dilutions onto cell cultures with further cytological (cytopathic effect and molecular confirmation (by BTV-NS1 group-specific PCR. The standardized technique was then applied to field samples (blood for detecting BTV. Results: The slope of the standard curve obtained was -3.23, and the efficiency was 103%. The LoD with RT-PCR was 8.269Ex103 number of copies of plasmid, whereas it was 13 with real-time PCR for plasmid dilutions. Similarly, LoD was determined for virus-spiked culture medium, and blood with both the types of PCR and the values were 103 TCID 50/ml and 104 TCID 50/ml with RT-PCR and 10° TCID 50/ml and 102 TCID 50/ml with real-time PCR, respectively. The standardized technique was applied to blood samples collected from BTV suspected animals; 10 among 20 samples were found positive with Cq values ranging from 27 to 39. The Cq value exhibiting samples were further processed in cell cultures and were confirmed to be BT positive. Likewise, Cq undetected samples on processing in cell cultures turned out to be BTV negative. Conclusion: Real-time PCR was found to be a very sensitive as well as reliable method

  12. Optimization of Critical Hairpin Features Allows miRNA-based Gene Knockdown Upon Single-copy Transduction

    Directory of Open Access Journals (Sweden)

    Renier Myburgh

    2014-01-01

    Full Text Available Gene knockdown using micro RNA (miRNA-based vector constructs is likely to become a prominent gene therapy approach. It was the aim of this study to improve the efficiency of gene knockdown through optimizing the structure of miRNA mimics. Knockdown of two target genes was analyzed: CCR5 and green fluorescent protein. We describe here a novel and optimized miRNA mimic design called mirGE comprising a lower stem length of 13 base pairs (bp, positioning of the targeting strand on the 5′ side of the miRNA, together with nucleotide mismatches in upper stem positions 1 and 12 placed on the passenger strand. Our mirGE proved superior to miR-30 in four aspects: yield of targeting strand incorporation into RNA-induced silencing complex (RISC; incorporation into RISC of correct targeting strand; precision of cleavage by Drosha; and ratio of targeting strand over passenger strand. A triple mirGE hairpin cassette targeting CCR5 was constructed. It allowed CCR5 knockdown with an efficiency of over 90% upon single-copy transduction. Importantly, single-copy expression of this construct rendered transduced target cells, including primary human macrophages, resistant to infection with a CCR5-tropic strain of HIV. Our results provide new insights for a better knockdown efficiency of constructs containing miRNA. Our results also provide the proof-of-principle that cells can be rendered HIV resistant through single-copy vector transduction, rendering this approach more compatible with clinical applications.

  13. Exploiting a Reference Genome in Terms of Duplications: The Network of Paralogs and Single Copy Genes in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Mara Sangiovanni

    2013-12-01

    Full Text Available Arabidopsis thaliana became the model organism for plant studies because of its small diploid genome, rapid lifecycle and short adult size. Its genome was the first among plants to be sequenced, becoming the reference in plant genomics. However, the Arabidopsis genome is characterized by an inherently complex organization, since it has undergone ancient whole genome duplications, followed by gene reduction, diploidization events and extended rearrangements, which relocated and split up the retained portions. These events, together with probable chromosome reductions, dramatically increased the genome complexity, limiting its role as a reference. The identification of paralogs and single copy genes within a highly duplicated genome is a prerequisite to understand its organization and evolution and to improve its exploitation in comparative genomics. This is still controversial, even in the widely studied Arabidopsis genome. This is also due to the lack of a reference bioinformatics pipeline that could exhaustively identify paralogs and singleton genes. We describe here a complete computational strategy to detect both duplicated and single copy genes in a genome, discussing all the methodological issues that may strongly affect the results, their quality and their reliability. This approach was used to analyze the organization of Arabidopsis nuclear protein coding genes, and besides classifying computationally defined paralogs into networks and single copy genes into different classes, it unraveled further intriguing aspects concerning the genome annotation and the gene relationships in this reference plant species. Since our results may be useful for comparative genomics and genome functional analyses, we organized a dedicated web interface to make them accessible to the scientific community.

  14. Robust synthesis for real-time systems

    DEFF Research Database (Denmark)

    Larsen, Kim Guldstrand; Legay, Axel; Traonouez, Luois-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...... of introducing small perturbations into formal models. We address this problem of robust implementations in timed specification theories. We first consider a fixed perturbation and study the robustness of timed specifications with respect to the operators of the theory. To this end we synthesize robust...... specification to an implementation, we need to reason about the possibility to effectively implement the theoretical specifications on physical systems, despite their limited precision. In the literature, this implementation problem has been linked to the robustness problem that analyzes the consequences...

  15. Real time speech formant analyzer and display

    Science.gov (United States)

    Holland, George E.; Struve, Walter S.; Homer, John F.

    1987-01-01

    A speech analyzer for interpretation of sound includes a sound input which converts the sound into a signal representing the sound. The signal is passed through a plurality of frequency pass filters to derive a plurality of frequency formants. These formants are converted to voltage signals by frequency-to-voltage converters and then are prepared for visual display in continuous real time. Parameters from the inputted sound are also derived and displayed. The display may then be interpreted by the user. The preferred embodiment includes a microprocessor which is interfaced with a television set for displaying of the sound formants. The microprocessor software enables the sound analyzer to present a variety of display modes for interpretive and therapeutic used by the user.

  16. CONSIDERATIONS ON REAL TIME DATA WAREHOUSING (RTDW

    Directory of Open Access Journals (Sweden)

    Marius Bogdan DINU

    2014-05-01

    Full Text Available The RTDW concept originated in the early 2000s. By that time, computing power had increased to a level that was allowing extraction of data collections for reporting purposes. Such collections were used almost in real time and at speeds nearly comparable to what an operation system was capable to deliver. The main idea will be to eliminate some of the components of the classic extraction process which is basically the most costly factor less time - consuming. We anticipate that the following factors will be decisive: elimination of batch-type processes [1], data compression techniques, data capture techniques, ability to keep in cache a large volume of data, parallel processing, and data mining algorithms that can adapt to such applications.

  17. Real-Time Optical Antimicrobial Susceptibility Testing

    DEFF Research Database (Denmark)

    Fredborg, Marlene; Andersen, Klaus R; Jørgensen, Erik

    2013-01-01

    Rapid antibiotic susceptibility testing is in highly demand in health-care fields as antimicrobial resistant bacterial strains emerge and spread. Here we describe an optical screening system (oCelloScope), which based on time-lapse imaging of 96 bacteria-antibiotic combinations at a time......, introduces real-time detection of bacterial growth and antimicrobial susceptibility, with imaging material to support the automatically generated graphs. Automated antibiotic susceptibility tests of a monoculture showed statistically significant antibiotic effect within 6 minutes and within 30 minutes...... from multidrug-resistant pathogenic bacteria. The oCelloScope system can be employed for a broad range of applications within bacteriology and may present new vistas as a point-of-care instrument in both clinical and veterinarian settings....

  18. Operational and real-time Business Intelligence

    Directory of Open Access Journals (Sweden)

    Daniela Ioana SANDU

    2008-01-01

    Full Text Available A key component of a company’s IT framework is a business intelligence (BI system. BI enables business users to report on, analyze and optimize business operations to reduce costs and increase revenues. Organizations use BI for strategic and tactical decision making where the decision-making cycle may span a time period of several weeks (e.g., campaign management or months (e.g., improving customer satisfaction.Competitive pressures coming from a very dynamic business environment are forcing companies to react faster to changing business conditions and customer requirements. As a result, there is now a need to use BI to help drive and optimize business operations on a daily basis, and, in some cases, even for intraday decision making. This type of BI is usually called operational business intelligence and real-time business intelligence.

  19. Performance evaluation of real time radiographic systems

    International Nuclear Information System (INIS)

    Venkatraman, B.; Saravanan, S.; Jayakumar, T.; Kalyanasundaram, P.; Baldev Raj

    1996-01-01

    The Real Time Radiography (RTR) system can be studied completely by knowing the modulation transfer function (MTF) of the whole system. The MTF curve is a special form of contrast/detail-size diagram in which the image contrast is plotted against the spatial frequency of a test object measured in line-pairs per millimetre (lp/mm). MTF curves are widely used to measure the characteristics of optical equipment, particularly for assessing the contribution of individual items in a complex imaging transfer system. Codes of practice indicate that the image intensifier systems should be checked periodically to assess its performance through the use of MTF curves and step wedges for contrast ratio. Authors, instead, suggest the use of performance curves which are simple to obtain and can be easily interpreted by radiographers. (author)

  20. Real time ray tracing based on shader

    Science.gov (United States)

    Gui, JiangHeng; Li, Min

    2017-07-01

    Ray tracing is a rendering algorithm for generating an image through tracing lights into an image plane, it can simulate complicate optical phenomenon like refraction, depth of field and motion blur. Compared with rasterization, ray tracing can achieve more realistic rendering result, however with greater computational cost, simple scene rendering can consume tons of time. With the GPU's performance improvement and the advent of programmable rendering pipeline, complicated algorithm can also be implemented directly on shader. So, this paper proposes a new method that implement ray tracing directly on fragment shader, mainly include: surface intersection, importance sampling and progressive rendering. With the help of GPU's powerful throughput capability, it can implement real time rendering of simple scene.

  1. Kalman Filtering with Real-Time Applications

    CERN Document Server

    Chui, Charles K

    2009-01-01

    Kalman Filtering with Real-Time Applications presents a thorough discussion of the mathematical theory and computational schemes of Kalman filtering. The filtering algorithms are derived via different approaches, including a direct method consisting of a series of elementary steps, and an indirect method based on innovation projection. Other topics include Kalman filtering for systems with correlated noise or colored noise, limiting Kalman filtering for time-invariant systems, extended Kalman filtering for nonlinear systems, interval Kalman filtering for uncertain systems, and wavelet Kalman filtering for multiresolution analysis of random signals. Most filtering algorithms are illustrated by using simplified radar tracking examples. The style of the book is informal, and the mathematics is elementary but rigorous. The text is self-contained, suitable for self-study, and accessible to all readers with a minimum knowledge of linear algebra, probability theory, and system engineering.

  2. Optimal, real-time control--colliders

    International Nuclear Information System (INIS)

    Spencer, J.E.

    1991-05-01

    With reasonable definitions, optimal control is possible for both classical and quantal systems with new approaches called PISC(Parallel) and NISC(Neural) from analogy with RISC (Reduced Instruction Set Computing). If control equals interaction, observation and comparison to some figure of merit with interaction via external fields, then optimization comes from varying these fields to give design or operating goals. Structural stability can then give us tolerance and design constraints. But simulations use simplified models, are not in real-time and assume fixed or stationary conditions, so optimal control goes far beyond convergence rates of algorithms. It is inseparable from design and this has many implications for colliders. 12 refs., 3 figs

  3. Real-time rockmass response from microseismics

    Energy Technology Data Exchange (ETDEWEB)

    Andrew King; Michael Lofgren; Matt van de Werken [CSIRO Exploration and Mining (Australia)

    2009-06-15

    The primary objective of this project was to develop a prototype real-time microseismic monitoring system for strata control management and forewarning of geotechnical hazards. Power and communications problems have been addressed by developing a wirelessly connected network of solar-powered acquisition nodes, one at the top of each instrumented borehole. The open-source 'earthworm' earthquake acquisition software, which can run on different hardware platforms and use different acquisition cards, was modified for use in a coal environment by developing special new arrival-picking and event-location procedures. The system was field-trialled at Moranbah North mine. The acquisition software performed well, as did wireless communications and solar power. There were issues with the acquisition hardware selected, including problems with timing synchronisation, which is essential for seismic event location. Although these were fixed during the test, different hardware is likely to be used in future installations.

  4. 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)

  5. Development of the real time monitor system

    Energy Technology Data Exchange (ETDEWEB)

    Kato, Katsumi [Research Organization for Information Science and Technology, Tokai, Ibaraki (Japan); Watanabe, Tadashi; Kaburaki, Hideo

    1996-10-01

    Large-scale simulation technique is studied at the Center for Promotion of Computational Science and Engineering (CCSE) for the computational science research in nuclear fields. Visualization and animation processing technique are studied and developed for efficient understanding of simulation results. The real time monitor system, in which on-going simulation results are transferred from a supercomputer or workstation to a graphic workstation and are visualized and recorded, is described in this report. This system is composed of the graphic workstation and the video equipment connected to the network. The control shell programs are the job-execution shell for simulations on supercomputers, the file-transfer shell for output files for visualization, and the shell for starting visualization tools. Special image processing technique and hardware are not necessary in this system and the standard visualization tool AVS and the UNIX commands are used, so that this system can be implemented and applied in various computer environments. (author)

  6. Embedded and real-time operating systems

    CERN Document Server

    Wang, K C

    2017-01-01

    This book covers the basic concepts and principles of operating systems, showing how to apply them to the design and implementation of complete operating systems for embedded and real-time systems. It includes all the foundational and background information on ARM architecture, ARM instructions and programming, toolchain for developing programs, virtual machines for software implementation and testing, program execution image, function call conventions, run-time stack usage and link C programs with assembly code. It describes the design and implementation of a complete OS for embedded systems in incremental steps, explaining the design principles and implementation techniques. For Symmetric Multiprocessing (SMP) embedded systems, the author examines the ARM MPcore processors, which include the SCU and GIC for interrupts routing and interprocessor communication and synchronization by Software Generated Interrupts (SGIs). Throughout the book, complete working sample systems demonstrate the design principles and...

  7. Real-time Human Activity Recognition

    Science.gov (United States)

    Albukhary, N.; Mustafah, Y. M.

    2017-11-01

    The traditional Closed-circuit Television (CCTV) system requires human to monitor the CCTV for 24/7 which is inefficient and costly. Therefore, there’s a need for a system which can recognize human activity effectively in real-time. This paper concentrates on recognizing simple activity such as walking, running, sitting, standing and landing by using image processing techniques. Firstly, object detection is done by using background subtraction to detect moving object. Then, object tracking and object classification are constructed so that different person can be differentiated by using feature detection. Geometrical attributes of tracked object, which are centroid and aspect ratio of identified tracked are manipulated so that simple activity can be detected.

  8. The RHIC real time data link system

    International Nuclear Information System (INIS)

    Hartmann, H.

    1997-01-01

    The RHIC Real Time Data Link (RTDL) System distributes to all locations around the RHIC ring machine parameters of general interest to accelerator systems and users. The system, along with supporting host interface, is centrally located. The RTDL System is comprised of two module types: the Encoder Module (V105) and the Input Module (V106). There is only one V105 module, but many (up to 128) Input Modules. Multiple buffered outputs are provided for use locally or for retransmission to other RHIC equipment locations. Machine parameters are generated from the V115 Waveform Generator Module (WFG) or from machine hardware and coupled directly through a fiber optic serial link to one of the V106 input channels

  9. REAL TIME DATA FOR REMEDIATION ACTIVITIES (11505)

    International Nuclear Information System (INIS)

    Brock, C.T.

    2011-01-01

    Health physicists from the CH2M HILL Plateau Remediation Company collaborated with Berkeley Nucleonics Corporation to modify the SAM 940 isotope identifier instrument to be used for nuclear waste remediation. These modifications coupled with existing capabilities of the SAM 940 have proven to be invaluable during remediation activities, reducing disposal costs by allowing swift remediation of targeted areas that have been identified as having isotopes of concern (IOC), and eliminating multiple visits to sites by declaring an excavation site clear of IOCs before demobilizing from the site. These advantages are enabled by accumulating spectral data for specific isotopes that is nearly 100 percent free of false positives, which are filtered out in 'real time.'

  10. A distributed real-time operating system

    International Nuclear Information System (INIS)

    Tuynman, F.; Hertzberger, L.O.

    1984-07-01

    A distributed real-time operating system, Fados, has been developed for an embedded multi-processor system. The operating system is based on a host target approach and provides for communication between arbitrary processes on host and target machine. The facilities offered are, apart from process communication, access to the file system on the host by programs on the target machine and monitoring and debugging of programs on the target machine from the host. The process communication has been designed in such a way that the possibilities are the same as those offered by the Ada programming language. The operating system is implemented on a MC 68000 based multiprocessor system in combination with a Unix host. (orig.)

  11. REAL TIME SPEED ESTIMATION FROM MONOCULAR VIDEO

    Directory of Open Access Journals (Sweden)

    M. S. Temiz

    2012-07-01

    Full Text Available In this paper, detailed studies have been performed for developing a real time system to be used for surveillance of the traffic flow by using monocular video cameras to find speeds of the vehicles for secure travelling are presented. We assume that the studied road segment is planar and straight, the camera is tilted downward a bridge and the length of one line segment in the image is known. In order to estimate the speed of a moving vehicle from a video camera, rectification of video images is performed to eliminate the perspective effects and then the interest region namely the ROI is determined for tracking the vehicles. Velocity vectors of a sufficient number of reference points are identified on the image of the vehicle from each video frame. For this purpose sufficient number of points from the vehicle is selected, and these points must be accurately tracked on at least two successive video frames. In the second step, by using the displacement vectors of the tracked points and passed time, the velocity vectors of those points are computed. Computed velocity vectors are defined in the video image coordinate system and displacement vectors are measured by the means of pixel units. Then the magnitudes of the computed vectors in the image space are transformed to the object space to find the absolute values of these magnitudes. The accuracy of the estimated speed is approximately ±1 – 2 km/h. In order to solve the real time speed estimation problem, the authors have written a software system in C++ programming language. This software system has been used for all of the computations and test applications.

  12. Real-time visualization of joint cavitation.

    Directory of Open Access Journals (Sweden)

    Gregory N Kawchuk

    Full Text Available Cracking sounds emitted from human synovial joints have been attributed historically to the sudden collapse of a cavitation bubble formed as articular surfaces are separated. Unfortunately, bubble collapse as the source of joint cracking is inconsistent with many physical phenomena that define the joint cracking phenomenon. Here we present direct evidence from real-time magnetic resonance imaging that the mechanism of joint cracking is related to cavity formation rather than bubble collapse. In this study, ten metacarpophalangeal joints were studied by inserting the finger of interest into a flexible tube tightened around a length of cable used to provide long-axis traction. Before and after traction, static 3D T1-weighted magnetic resonance images were acquired. During traction, rapid cine magnetic resonance images were obtained from the joint midline at a rate of 3.2 frames per second until the cracking event occurred. As traction forces increased, real-time cine magnetic resonance imaging demonstrated rapid cavity inception at the time of joint separation and sound production after which the resulting cavity remained visible. Our results offer direct experimental evidence that joint cracking is associated with cavity inception rather than collapse of a pre-existing bubble. These observations are consistent with tribonucleation, a known process where opposing surfaces resist separation until a critical point where they then separate rapidly creating sustained gas cavities. Observed previously in vitro, this is the first in-vivo macroscopic demonstration of tribonucleation and as such, provides a new theoretical framework to investigate health outcomes associated with joint cracking.

  13. Real Time Monitor of Grid job executions

    International Nuclear Information System (INIS)

    Colling, D J; Martyniak, J; McGough, A S; Krenek, A; Sitera, J; Mulac, M; Dvorak, F

    2010-01-01

    In this paper we describe the architecture and operation of the Real Time Monitor (RTM), developed by the Grid team in the HEP group at Imperial College London. This is arguably the most popular dissemination tool within the EGEE [1] Grid. Having been used, on many occasions including GridFest and LHC inauguration events held at CERN in October 2008. The RTM gathers information from EGEE sites hosting Logging and Bookkeeping (LB) services. Information is cached locally at a dedicated server at Imperial College London and made available for clients to use in near real time. The system consists of three main components: the RTM server, enquirer and an apache Web Server which is queried by clients. The RTM server queries the LB servers at fixed time intervals, collecting job related information and storing this in a local database. Job related data includes not only job state (i.e. Scheduled, Waiting, Running or Done) along with timing information but also other attributes such as Virtual Organization and Computing Element (CE) queue - if known. The job data stored in the RTM database is read by the enquirer every minute and converted to an XML format which is stored on a Web Server. This decouples the RTM server database from the client removing the bottleneck problem caused by many clients simultaneously accessing the database. This information can be visualized through either a 2D or 3D Java based client with live job data either being overlaid on to a 2 dimensional map of the world or rendered in 3 dimensions over a globe map using OpenGL.

  14. Development of a primer–probe energy transfer based real-time PCR for the detection of Swine influenza virus

    DEFF Research Database (Denmark)

    Kowalczyk, Andrzej; Markowska-Daniel, Iwona; Rasmussen, Thomas Bruun

    2013-01-01

    Swine influenza virus (SIV) causes a contagious and requiring official notification disease of pigs and humans. In this study, a real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on primer–probe energy transfer (PriProET) for the detection of SIV RNA was developed...... of the specific product amplification. The assay is specific for influenza virus with a sensitivity of detection limit of approximately 10 copies of RNA by PCR. Based on serial dilutions of SIV, the detection limit of the assay was approximately 0.003 TCID50/ml for H1N1 A/Swine/Poland/KPR9/2004 virus. The Pri...

  15. Internal control for real-time polymerase chain reaction based on MS2 bacteriophage for RNA viruses diagnostics.

    Science.gov (United States)

    Zambenedetti, Miriam Ribas; Pavoni, Daniela Parada; Dallabona, Andreia Cristine; Dominguez, Alejandro Correa; Poersch, Celina de Oliveira; Fragoso, Stenio Perdigão; Krieger, Marco Aurélio

    2017-05-01

    Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. In Brazil, it is mandatory the use of nucleic acid tests to detect hepatitis C virus (HCV), hepatitis B virus and human immunodeficiency virus in blood banks because of the immunological window. The use of an internal control (IC) is necessary to differentiate the true negative results from those consequent from a failure in some step of the nucleic acid test. The aim of this study was the construction of virus-modified particles, based on MS2 bacteriophage, to be used as IC for the diagnosis of RNA viruses. The MS2 genome was cloned into the pET47b(+) plasmid, generating pET47b(+)-MS2. MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. These particles were used as non-competitive IC in assays for RNA virus diagnostics. In addition, a competitive control for HCV diagnosis was developed by cloning a mutated HCV sequence into the MS2 replicase gene of pET47b(+)-MS2, which produces a non-propagating MS2 particle. The utility of MS2-like particles as IC was evaluated in a one-step format multiplex real-time RT-PCR for HCV detection. We demonstrated that both competitive and non-competitive IC could be successfully used to monitor the HCV amplification performance, including the extraction, reverse transcription, amplification and detection steps, without compromising the detection of samples with low target concentrations. In conclusion, MS2-like particles generated by this strategy proved to be useful IC for RNA virus diagnosis, with advantage that they are produced by a low cost protocol. An attractive feature of this system is that it allows the construction of a multicontrol by the insertion of sequences from more than one pathogen, increasing its applicability for diagnosing different RNA viruses.

  16. Approaching near real-time biosensing: microfluidic microsphere based biosensor for real-time analyte detection.

    Science.gov (United States)

    Cohen, Noa; Sabhachandani, Pooja; Golberg, Alexander; Konry, Tania

    2015-04-15

    In this study we describe a simple lab-on-a-chip (LOC) biosensor approach utilizing well mixed microfluidic device and a microsphere-based assay capable of performing near real-time diagnostics of clinically relevant analytes such cytokines and antibodies. We were able to overcome the adsorption kinetics reaction rate-limiting mechanism, which is diffusion-controlled in standard immunoassays, by introducing the microsphere-based assay into well-mixed yet simple microfluidic device with turbulent flow profiles in the reaction regions. The integrated microsphere-based LOC device performs dynamic detection of the analyte in minimal amount of biological specimen by continuously sampling micro-liter volumes of sample per minute to detect dynamic changes in target analyte concentration. Furthermore we developed a mathematical model for the well-mixed reaction to describe the near real time detection mechanism observed in the developed LOC method. To demonstrate the specificity and sensitivity of the developed real time monitoring LOC approach, we applied the device for clinically relevant analytes: Tumor Necrosis Factor (TNF)-α cytokine and its clinically used inhibitor, anti-TNF-α antibody. Based on the reported results herein, the developed LOC device provides continuous sensitive and specific near real-time monitoring method for analytes such as cytokines and antibodies, reduces reagent volumes by nearly three orders of magnitude as well as eliminates the washing steps required by standard immunoassays. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Application of XML in real-time data warehouse

    Science.gov (United States)

    Zhao, Yanhong; Wang, Beizhan; Liu, Lizhao; Ye, Su

    2009-07-01

    At present, XML is one of the most widely-used technologies of data-describing and data-exchanging, and the needs for real-time data make real-time data warehouse a popular area in the research of data warehouse. What effects can we have if we apply XML technology to the research of real-time data warehouse? XML technology solves many technologic problems which are impossible to be addressed in traditional real-time data warehouse, and realize the integration of OLAP (On-line Analytical Processing) and OLTP (Online transaction processing) environment. Then real-time data warehouse can truly be called "real time".

  18. Real Time Seismic Prediction while Drilling

    Science.gov (United States)

    Schilling, F. R.; Bohlen, T.; Edelmann, T.; Kassel, A.; Heim, A.; Gehring, M.; Lüth, S.; Giese, R.; Jaksch, K.; Rechlin, A.; Kopf, M.; Stahlmann, J.; Gattermann, J.; Bruns, B.

    2009-12-01

    Efficient and safe drilling is a prerequisite to enhance the mobility of people and goods, to improve the traffic as well as utility infrastructure of growing megacities, and to ensure the growing energy demand while building geothermal and in hydroelectric power plants. Construction within the underground is often building within the unknown. An enhanced risk potential for people and the underground building may arise if drilling enters fracture zones, karsts, brittle rocks, mixed solid and soft rocks, caves, or anthropogenic obstacles. Knowing about the material behavior ahead of the drilling allows reducing the risk during drilling and construction operation. In drilling operations direct observations from boreholes can be complemented with geophysical investigations. In this presentation we focus on “real time” seismic prediction while drilling which is seen as a prerequisite while using geophysical methods in modern drilling operations. In solid rocks P- and S-wave velocity, refraction and reflection as well as seismic wave attenuation can be used for the interpretation of structures ahead of the drilling. An Integrated Seismic Imaging System (ISIS) for exploration ahead of a construction is used, where a pneumatic hammer or a magnetostrictive vibration source generate repetitive signals behind the tunneling machine. Tube waves are generated which travel along the tunnel to the working face. There the tube waves are converted to mainly S- but also P-Waves which interact with the formation ahead of the heading face. The reflected or refracted waves travel back to the working front are converted back to tube waves and recorded using three-component geophones which are fit into the tips of anchor rods. In near real time, the ISIS software allows for an integrated 3D imaging and interpretation of the observed data, geological and geotechnical parameters. Fracture zones, heterogeneities, and variations in the rock properties can be revealed during the drilling

  19. TTT and PIKK Complex Genes Reverted to Single Copy Following Polyploidization and Retain Function Despite Massive Retrotransposition in Maize.

    Science.gov (United States)

    Garcia, Nelson; Messing, Joachim

    2017-01-01

    The TEL2, TTI1, and TTI2 proteins are co-chaperones for heat shock protein 90 (HSP90) to regulate the protein folding and maturation of phosphatidylinositol 3-kinase-related kinases (PIKKs). Referred to as the TTT complex, the genes that encode them are highly conserved from man to maize. TTT complex and PIKK genes exist mostly as single copy genes in organisms where they have been characterized. Members of this interacting protein network in maize were identified and synteny analyses were performed to study their evolution. Similar to other species, there is only one copy of each of these genes in maize which was due to a loss of the duplicated copy created by ancient allotetraploidy. Moreover, the retained copies of the TTT complex and the PIKK genes tolerated extensive retrotransposon insertion in their introns that resulted in increased gene lengths and gene body methylation, without apparent effect in normal gene expression and function. The results raise an interesting question on whether the reversion to single copy was due to selection against deleterious unbalanced gene duplications between members of the complex as predicted by the gene balance hypothesis, or due to neutral loss of extra copies. Uneven alteration of dosage either by adding extra copies or modulating gene expression of complex members is being proposed as a means to investigate whether the data supports the gene balance hypothesis or not.

  20. TTT and PIKK Complex Genes Reverted to Single Copy Following Polyploidization and Retain Function Despite Massive Retrotransposition in Maize

    Directory of Open Access Journals (Sweden)

    Nelson Garcia

    2017-11-01

    Full Text Available The TEL2, TTI1, and TTI2 proteins are co-chaperones for heat shock protein 90 (HSP90 to regulate the protein folding and maturation of phosphatidylinositol 3-kinase-related kinases (PIKKs. Referred to as the TTT complex, the genes that encode them are highly conserved from man to maize. TTT complex and PIKK genes exist mostly as single copy genes in organisms where they have been characterized. Members of this interacting protein network in maize were identified and synteny analyses were performed to study their evolution. Similar to other species, there is only one copy of each of these genes in maize which was due to a loss of the duplicated copy created by ancient allotetraploidy. Moreover, the retained copies of the TTT complex and the PIKK genes tolerated extensive retrotransposon insertion in their introns that resulted in increased gene lengths and gene body methylation, without apparent effect in normal gene expression and function. The results raise an interesting question on whether the reversion to single copy was due to selection against deleterious unbalanced gene duplications between members of the complex as predicted by the gene balance hypothesis, or due to neutral loss of extra copies. Uneven alteration of dosage either by adding extra copies or modulating gene expression of complex members is being proposed as a means to investigate whether the data supports the gene balance hypothesis or not.

  1. Real Time Earthquake Information System in Japan

    Science.gov (United States)

    Doi, K.; Kato, T.

    2003-12-01

    An early earthquake notification system in Japan had been developed by the Japan Meteorological Agency (JMA) as a governmental organization responsible for issuing earthquake information and tsunami forecasts. The system was primarily developed for prompt provision of a tsunami forecast to the public with locating an earthquake and estimating its magnitude as quickly as possible. Years after, a system for a prompt provision of seismic intensity information as indices of degrees of disasters caused by strong ground motion was also developed so that concerned governmental organizations can decide whether it was necessary for them to launch emergency response or not. At present, JMA issues the following kinds of information successively when a large earthquake occurs. 1) Prompt report of occurrence of a large earthquake and major seismic intensities caused by the earthquake in about two minutes after the earthquake occurrence. 2) Tsunami forecast in around three minutes. 3) Information on expected arrival times and maximum heights of tsunami waves in around five minutes. 4) Information on a hypocenter and a magnitude of the earthquake, the seismic intensity at each observation station, the times of high tides in addition to the expected tsunami arrival times in 5-7 minutes. To issue information above, JMA has established; - An advanced nationwide seismic network with about 180 stations for seismic wave observation and about 3,400 stations for instrumental seismic intensity observation including about 2,800 seismic intensity stations maintained by local governments, - Data telemetry networks via landlines and partly via a satellite communication link, - Real-time data processing techniques, for example, the automatic calculation of earthquake location and magnitude, the database driven method for quantitative tsunami estimation, and - Dissemination networks, via computer-to-computer communications and facsimile through dedicated telephone lines. JMA operationally

  2. The INGV Real Time Strong Motion Database

    Science.gov (United States)

    Massa, Marco; D'Alema, Ezio; Mascandola, Claudia; Lovati, Sara; Scafidi, Davide; Gomez, Antonio; Carannante, Simona; Franceschina, Gianlorenzo; Mirenna, Santi; Augliera, Paolo

    2017-04-01

    The INGV real time strong motion data sharing is assured by the INGV Strong Motion Database. ISMD (http://ismd.mi.ingv.it) was designed in the last months of 2011 in cooperation among different INGV departments, with the aim to organize the distribution of the INGV strong-motion data using standard procedures for data acquisition and processing. The first version of the web portal was published soon after the occurrence of the 2012 Emilia (Northern Italy), Mw 6.1, seismic sequence. At that time ISMD was the first European real time web portal devoted to the engineering seismology community. After four years of successfully operation, the thousands of accelerometric waveforms collected in the archive need necessary a technological improvement of the system in order to better organize the new data archiving and to make more efficient the answer to the user requests. ISMD 2.0 was based on PostgreSQL (www.postgresql.org), an open source object- relational database. The main purpose of the web portal is to distribute few minutes after the origin time the accelerometric waveforms and related metadata of the Italian earthquakes with ML≥3.0. Data are provided both in raw SAC (counts) and automatically corrected ASCII (gal) formats. The web portal also provide, for each event, a detailed description of the ground motion parameters (i.e. Peak Ground Acceleration, Velocity and Displacement, Arias and Housner Intensities) data converted in velocity and displacement, response spectra up to 10.0 s and general maps concerning the recent and the historical seismicity of the area together with information about its seismic hazard. The focal parameters of the events are provided by the INGV National Earthquake Center (CNT, http://cnt.rm.ingv.it). Moreover, the database provides a detailed site characterization section for each strong motion station, based on geological, geomorphological and geophysical information. At present (i.e. January 2017), ISMD includes 987 (121

  3. RTMOD: Real-Time MODel evaluation

    International Nuclear Information System (INIS)

    Graziani, G; Galmarini, S.; Mikkelsen, T.

    2000-01-01

    The 1998 - 1999 RTMOD project is a system based on an automated statistical evaluation for the inter-comparison of real-time forecasts produced by long-range atmospheric dispersion models for national nuclear emergency predictions of cross-boundary consequences. The background of RTMOD was the 1994 ETEX project that involved about 50 models run in several Institutes around the world to simulate two real tracer releases involving a large part of the European territory. In the preliminary phase of ETEX, three dry runs (i.e. simulations in real-time of fictitious releases) were carried out. At that time, the World Wide Web was not available to all the exercise participants, and plume predictions were therefore submitted to JRC-Ispra by fax and regular mail for subsequent processing. The rapid development of the World Wide Web in the second half of the nineties, together with the experience gained during the ETEX exercises suggested the development of this project. RTMOD featured a web-based user-friendly interface for data submission and an interactive program module for displaying, intercomparison and analysis of the forecasts. RTMOD has focussed on model intercomparison of concentration predictions at the nodes of a regular grid with 0.5 degrees of resolution both in latitude and in longitude, the domain grid extending from 5W to 40E and 40N to 65N. Hypothetical releases were notified around the world to the 28 model forecasters via the web on a one-day warning in advance. They then accessed the RTMOD web page for detailed information on the actual release, and as soon as possible they then uploaded their predictions to the RTMOD server and could soon after start their inter-comparison analysis with other modelers. When additional forecast data arrived, already existing statistical results would be recalculated to include the influence by all available predictions. The new web-based RTMOD concept has proven useful as a practical decision-making tool for realtime

  4. Business Hypervisors for Real-time Applications

    Directory of Open Access Journals (Sweden)

    L. Perneel

    2015-08-01

    Full Text Available System virtualization is one of the hottest trends in information technology today. It is not just another nice to use technology but has become fundamental across the business world. It is successfully used with many business application classes where cloud computing is the most visual one. Recently, it started to be used for soft Real-Time (RT applications such as IP telephony, media servers, audio and video streaming servers, automotive and communication systems in general. Running these applications on a traditional system (Hardware + Operating System guarantee their Quality of Service (QoS; virtualizing them means inserting a new layer between the hardware and the (virtual Operating System (OS, and thus adding extra overhead. Although these applications’ areas do not always demand hard time guarantees, they require the underlying virtualization layer supports low latency and provide adequate computational resources for completion within a reasonable or predictable timeframe. These aspects are intimately intertwined with the logic of the hypervisor scheduler. In this paper, a series of tests are conducted on three hypervisors (VMware ESXi, Hyper-V server and Xen to provide a benchmark of the latencies added to the applications running on top of them. These tests are conducted for different scenarios (use cases to take into consideration all the parameters and configurations of the hypervisors’ schedulers. Finally, this benchmark can be used as a reference for choosing the best hypervisor-application combination.

  5. Real time biometric surveillance with gait recognition

    Science.gov (United States)

    Mohapatra, Subasish; Swain, Anisha; Das, Manaswini; Mohanty, Subhadarshini

    2018-04-01

    Bio metric surveillance has become indispensable for every system in the recent years. The contribution of bio metric authentication, identification, and screening purposes are widely used in various domains for preventing unauthorized access. A large amount of data needs to be updated, segregated and safeguarded from malicious software and misuse. Bio metrics is the intrinsic characteristics of each individual. Recently fingerprints, iris, passwords, unique keys, and cards are commonly used for authentication purposes. These methods have various issues related to security and confidentiality. These systems are not yet automated to provide the safety and security. The gait recognition system is the alternative for overcoming the drawbacks of the recent bio metric based authentication systems. Gait recognition is newer as it hasn't been implemented in the real-world scenario so far. This is an un-intrusive system that requires no knowledge or co-operation of the subject. Gait is a unique behavioral characteristic of every human being which is hard to imitate. The walking style of an individual teamed with the orientation of joints in the skeletal structure and inclinations between them imparts the unique characteristic. A person can alter one's own external appearance but not skeletal structure. These are real-time, automatic systems that can even process low-resolution images and video frames. In this paper, we have proposed a gait recognition system and compared the performance with conventional bio metric identification systems.

  6. The Colliderscope: a real-time show

    CERN Multimedia

    Francesco Poppi

    2010-01-01

    Ninety-six LED lights distributed over the facade of the Niels Bohr Institute (NBI) in Blegdamsvej (Denmark) reproduce the actual signals coming from the Transition Radiation Detector (TRT) in ATLAS. Thanks to the Colliderscope, when a collision occurs below the ground in Geneva, people passing by in Blegdamsvej will be aware of it almost in real-time.   Niels Bohr Institute facade lit up to reflect the latest data from ATLAS-TRT . The pattern, intensity and duration of the Colliderscope’s flashes of light depend on the physical parameters of particles crossing the ATLAS TRT detector. “At the Colliderscope very little happens randomly”, explains Troels Petersen, a physicist at NBI and one of the people who conceived it. “Particularly interesting events, such as electrons, are shown by a bright light that remains on the facade for several seconds”. The Niels Bohr Institute has participated in the development of the TRT detector, and this is why t...

  7. Real-Time 3D Visualization

    Science.gov (United States)

    1997-01-01

    Butler Hine, former director of the Intelligent Mechanism Group (IMG) at Ames Research Center, and five others partnered to start Fourth Planet, Inc., a visualization company that specializes in the intuitive visual representation of dynamic, real-time data over the Internet and Intranet. Over a five-year period, the then NASA researchers performed ten robotic field missions in harsh climes to mimic the end- to-end operations of automated vehicles trekking across another world under control from Earth. The core software technology for these missions was the Virtual Environment Vehicle Interface (VEVI). Fourth Planet has released VEVI4, the fourth generation of the VEVI software, and NetVision. VEVI4 is a cutting-edge computer graphics simulation and remote control applications tool. The NetVision package allows large companies to view and analyze in virtual 3D space such things as the health or performance of their computer network or locate a trouble spot on an electric power grid. Other products are forthcoming. Fourth Planet is currently part of the NASA/Ames Technology Commercialization Center, a business incubator for start-up companies.

  8. Real-time control of fusion reactors

    International Nuclear Information System (INIS)

    Goncalves, B.; Sousa, J.; Varandas, C.A.F.

    2010-01-01

    The next generation fusion experiments, e.g. ITER, will be highly complex and raise new challenges in the field of control and data acquisition systems. The more advanced operation scenarios have to be capable of sustaining long pulse steady-state plasma and to suppress plasma instabilities almost completely. Such scenarios will heavily rely on Multiple-Input-Multiple-Output (MIMO) fast control systems. To ensure safety for the operation these systems have to be robust and resilient to faults while ensuring high availability. Mindful of the importance of such features for future fusion experiments ATCA based systems have been successfully used in fusion experiment as MIMO fast controller. This is the most promising architecture to substantially enhance the performance and capability of existing standard systems delivering well high throughput as well as high availability. The real-time control needs of a fusion experiment, the rational for the presently pursued solutions, the existing problems and the broad scientific and technical questions that need to be addressed on the path to a fusion power plant will be discussed.

  9. Real-time petroleum spill detection system

    International Nuclear Information System (INIS)

    Dakin, D.T.

    2001-01-01

    A real-time autonomous oil and fuel spill detection system has been developed to rapidly detect of a wide range of petroleum products floating on, or suspended in water. The system consists of an array of spill detection buoys distributed within the area to be monitored. The buoys are composed of a float and a multispectral fluorometer, which looks up through the top 5 cm of water to detect floating and suspended petroleum products. The buoys communicate to a base station computer that controls the sampling of the buoys and analyses the data from each buoy to determine if a spill has occurred. If statistically significant background petroleum levels are detected, the system raises an oil spill alarm. The system is useful because early detection of a marine oil spill allows for faster containment, thereby minimizing the contaminated area and reducing cleanup costs. This paper also provided test results for biofouling, various petroleum product detection, water turbidity and wave tolerance. The technology has been successfully demonstrated. The UV light source keeps the optic window free from biofouling, and the electronics are fully submerged so there is no risk that the unit could ignite the vapours of a potential oil spill. The system can also tolerate moderately turbid waters and can therefore be used in many rivers, harbours, water intakes and sumps. The system can detect petroleum products with an average thickness of less than 3 micrometers floating on the water surface. 3 refs., 15 figs

  10. Real-time ultrasonic weld evaluation system

    Science.gov (United States)

    Katragadda, Gopichand; Nair, Satish; Liu, Harry; Brown, Lawrence M.

    1996-11-01

    Ultrasonic testing techniques are currently used as an alternative to radiography for detecting, classifying,and sizing weld defects, and for evaluating weld quality. Typically, ultrasonic weld inspections are performed manually, which require significant operator expertise and time. Thus, in recent years, the emphasis is to develop automated methods to aid or replace operators in critical weld inspections where inspection time, reliability, and operator safety are major issues. During this period, significant advances wee made in the areas of weld defect classification and sizing. Very few of these methods, however have found their way into the market, largely due to the lack of an integrated approach enabling real-time implementation. Also, not much research effort was directed in improving weld acceptance criteria. This paper presents an integrated system utilizing state-of-the-art techniques for a complete automation of the weld inspection procedure. The modules discussed include transducer tracking, classification, sizing, and weld acceptance criteria. Transducer tracking was studied by experimentally evaluating sonic and optical position tracking techniques. Details for this evaluation are presented. Classification is obtained using a multi-layer perceptron. Results from different feature extraction schemes, including a new method based on a combination of time and frequency-domain signal representations are given. Algorithms developed to automate defect registration and sizing are discussed. A fuzzy-logic acceptance criteria for weld acceptance is presented describing how this scheme provides improved robustness compared to the traditional flow-diagram standards.

  11. Real-Time Accumulative Computation Motion Detectors

    Directory of Open Access Journals (Sweden)

    Saturnino Maldonado-Bascón

    2009-12-01

    Full Text Available The neurally inspired accumulative computation (AC method and its application to motion detection have been introduced in the past years. This paper revisits the fact that many researchers have explored the relationship between neural networks and finite state machines. Indeed, finite state machines constitute the best characterized computational model, whereas artificial neural networks have become a very successful tool for modeling and problem solving. The article shows how to reach real-time performance after using a model described as a finite state machine. This paper introduces two steps towards that direction: (a A simplification of the general AC method is performed by formally transforming it into a finite state machine. (b A hardware implementation in FPGA of such a designed AC module, as well as an 8-AC motion detector, providing promising performance results. We also offer two case studies of the use of AC motion detectors in surveillance applications, namely infrared-based people segmentation and color-based people tracking, respectively.

  12. Mobility and language change in real time

    DEFF Research Database (Denmark)

    Monka, Malene

    Diachronic studies of the interrelationship between mobility and language change leave us with some unanswered questions of causation. The most important question is whether language change is caused by mobility, or if mobile informants mark themselves linguistically different than their non-mobi...... mobile and non-mobile informants. I also suggest a human geographic approach to place to explain the differences between the language change of the mobile informants (e.g. Britain 2009; Johnstone 2004).......Diachronic studies of the interrelationship between mobility and language change leave us with some unanswered questions of causation. The most important question is whether language change is caused by mobility, or if mobile informants mark themselves linguistically different than their non......-mobile peers prior to being geographically and socially mobile (e.g. Andersson & Thelander 1994). In the presentation I discuss this question by presenting a real time panel-study of language change in 23 speakers from three municipalities in distinct dialect areas in Denmark. The language change of six mobile...

  13. The Fast Tracker Real Time Processor

    CERN Document Server

    Annovi, A; The ATLAS collaboration

    2011-01-01

    As the LHC luminosity is ramped up to the SLHC Phase I level and beyond, the high rates, multiplicities, and energies of particles seen by the detectors will pose a unique challenge. Only a tiny fraction of the produced collisions can be stored on tape and immense real-time data reduction is needed. An effective trigger system must maintain high trigger efficiencies for the physics we are most interested in, and at the same time suppress the enormous QCD backgrounds. This requires massive computing power to minimize the online execution time of complex algorithms. A multi-level trigger is an effective solution for an otherwise impossible problem. The Fast Tracker (FTK)[1], is a proposed upgrade to the current ATLAS trigger system that will operate at full Level-1 output rates and provide high quality tracks reconstructed over the entire detector by the start of processing in Level-2. FTK solves the combinatorial challenge inherent to tracking by exploiting massive parallelism of associative memories [2] that ...

  14. Near Real Time Ship Detection Experiments

    Science.gov (United States)

    Brusch, S.; Lehner, S.; Schwarz, E.; Fritz, T.

    2010-04-01

    A new Near Real Time (NRT) ship detection processor SAINT (SAR AIS Integrated Toolbox) was developed in the framework of the ESA project MARISS. Data are received at DLRs ground segment DLR-BN (Neustrelitz, Germany). Results of the ship detection are available on ftp server within 30 min after the acquisition started. The detectability of ships on Synthetic Aperture Radar (SAR) ERS-2, ENVISAT ASAR and TerraSAR-X (TS-X) images is validated by coastal (live) AIS and space AIS. The monitoring areas chosen for surveillance are the North-, Baltic Sea, and Cape Town. The detectability in respect to environmental parameters like wind field, sea state, currents and changing coastlines due to tidal effects is investigated. In the South Atlantic a tracking experiment of the German research vessel Polarstern has been performed. Issues of piracy in particular in respect to ships hijacked at the Somali coast are discussed. Some examples using high resolution images from TerraSAR-X are given.

  15. Real-time scheduling of software tasks

    International Nuclear Information System (INIS)

    Hoff, L.T.

    1995-01-01

    When designing real-time systems, it is often desirable to schedule execution of software tasks based on the occurrence of events. The events may be clock ticks, interrupts from a hardware device, or software signals from other software tasks. If the nature of the events, is well understood, this scheduling is normally a static part of the system design. If the nature of the events is not completely understood, or is expected to change over time, it may be necessary to provide a mechanism for adjusting the scheduling of the software tasks. RHIC front-end computers (FECs) provide such a mechanism. The goals in designing this mechanism were to be as independent as possible of the underlying operating system, to allow for future expansion of the mechanism to handle new types of events, and to allow easy configuration. Some considerations which steered the design were programming paradigm (object oriented vs. procedural), programming language, and whether events are merely interesting moments in time, or whether they intrinsically have data associated with them. The design also needed to address performance and robustness tradeoffs involving shared task contexts, task priorities, and use of interrupt service routine (ISR) contexts vs. task contexts. This paper will explore these considerations and tradeoffs

  16. An improved real time superresolution FPGA system

    Science.gov (United States)

    Lakshmi Narasimha, Pramod; Mudigoudar, Basavaraj; Yue, Zhanfeng; Topiwala, Pankaj

    2009-05-01

    In numerous computer vision applications, enhancing the quality and resolution of captured video can be critical. Acquired video is often grainy and low quality due to motion, transmission bottlenecks, etc. Postprocessing can enhance it. Superresolution greatly decreases camera jitter to deliver a smooth, stabilized, high quality video. In this paper, we extend previous work on a real-time superresolution application implemented in ASIC/FPGA hardware. A gradient based technique is used to register the frames at the sub-pixel level. Once we get the high resolution grid, we use an improved regularization technique in which the image is iteratively modified by applying back-projection to get a sharp and undistorted image. The algorithm was first tested in software and migrated to hardware, to achieve 320x240 -> 1280x960, about 30 fps, a stunning superresolution by 16X in total pixels. Various input parameters, such as size of input image, enlarging factor and the number of nearest neighbors, can be tuned conveniently by the user. We use a maximum word size of 32 bits to implement the algorithm in Matlab Simulink as well as in FPGA hardware, which gives us a fine balance between the number of bits and performance. The proposed system is robust and highly efficient. We have shown the performance improvement of the hardware superresolution over the software version (C code).

  17. Optimizing near real time accountability for reprocessing

    International Nuclear Information System (INIS)

    Cipiti, Benjamin B.

    2010-01-01

    Near Real Time Accountability (NRTA) of actinides at high precision in reprocessing plants has been a long sought-after goal in the safeguards community. Achieving this goal is hampered by the difficulty of making precision measurements in the reprocessing environment, equipment cost, and impact to plant operations. Thus the design of future reprocessing plants requires an optimization of different approaches. The Separations and Safeguards Performance Model, developed at Sandia National Laboratories, was used to evaluate a number of NRTA strategies in a UREX+ reprocessing plant. Strategies examined include the incorporation of additional actinide measurements of internal plant vessels, more use of process monitoring data, and the option of periodic draining of inventory to key tanks. Preliminary results show that the addition of measurement technologies can increase the overall measurement uncertainty due to additional error propagation, so care must be taken when designing an advanced system. Initial results also show that relying on a combination of different NRTA techniques will likely be the best option. The model provides a platform for integrating all the data. The modeling results for the different NRTA options under various material loss conditions will be presented.

  18. Improved Detection of Lassa Virus by Reverse Transcription-PCR Targeting the 5′ Region of S RNA▿

    OpenAIRE

    Ölschläger, Stephan; Lelke, Michaela; Emmerich, Petra; Panning, Marcus; Drosten, Christian; Hass, Meike; Asogun, Danny; Ehichioya, Deborah; Omilabu, Sunday; Günther, Stephan

    2010-01-01

    The method of choice for the detection of Lassa virus is reverse transcription (RT)-PCR. However, the high degree of genetic variability of the virus poses a problem with the design of RT-PCR assays that will reliably detect all strains. Recently, we encountered difficulties in detecting some strains from Liberia and Nigeria in a commonly used glycoprotein precursor (GPC) gene-specific RT-PCR assay (A. H. Demby, J. Chamberlain, D. W. Brown, and C. S. Clegg, J. Clin. Microbiol. 32:2898-2903, 1...

  19. Rapid detection of genetically diverse tomato black ring virus isolates using reverse transcription loop-mediated isothermal amplification.

    Science.gov (United States)

    Hasiów-Jaroszewska, Beata; Budzyńska, Daria; Borodynko, Natasza; Pospieszny, Henryk

    2015-12-01

    A reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) has been developed for detection of tomato black ring virus (TBRV) isolates collected from different hosts. One-step RT-LAMP was performed with a set of four primers, the design of which was based on the coat protein gene. Results of RT-LAMP were visualized by direct staining of products with fluorescent dyes, agarose gel electrophoresis, and analysis of amplification curves. The sensitivity of RT-LAMP was 100-fold greater than that of RT-PCR. The RT-LAMP assay developed here is a useful and practical method for diagnosis of TBRV.

  20. Microdroplet sandwich real-time rt-PCR for detection of pandemic and seasonal influenza subtypes.

    Directory of Open Access Journals (Sweden)

    Stephanie L Angione

    Full Text Available As demonstrated by the recent 2012/2013 flu epidemic, the continual emergence of new viral strains highlights the need for accurate medical diagnostics in multiple community settings. If rapid, robust, and sensitive diagnostics for influenza subtyping were available, it would help identify epidemics, facilitate appropriate antiviral usage, decrease inappropriate antibiotic usage, and eliminate the extra cost of unnecessary laboratory testing and treatment. Here, we describe a droplet sandwich platform that can detect influenza subtypes using real-time reverse-transcription polymerase chain reaction (rtRT-PCR. Using clinical samples collected during the 2010/11 season, we effectively differentiate between H1N1p (swine pandemic, H1N1s (seasonal, and H3N2 with an overall assay sensitivity was 96%, with 100% specificity for each subtype. Additionally, we demonstrate the ability to detect viral loads as low as 10(4 copies/mL, which is two orders of magnitude lower than viral loads in typical infected patients. This platform performs diagnostics in a miniaturized format without sacrificing any sensitivity, and can thus be easily developed into devices which are ideal for small clinics and pharmacies.

  1. Identification of Reference Genes for Normalizing Quantitative Real-Time PCR in Urechis unicinctus

    Science.gov (United States)

    Bai, Yajiao; Zhou, Di; Wei, Maokai; Xie, Yueyang; Gao, Beibei; Qin, Zhenkui; Zhang, Zhifeng

    2018-06-01

    The reverse transcription quantitative real-time PCR (RT-qPCR) has become one of the most important techniques of studying gene expression. A set of valid reference genes are essential for the accurate normalization of data. In this study, five candidate genes were analyzed with geNorm, NormFinder, BestKeeper and ΔCt methods to identify the genes stably expressed in echiuran Urechis unicinctus, an important commercial marine benthic worm, under abiotic (sulfide stress) and normal (adult tissues, embryos and larvae at different development stages) conditions. The comprehensive results indicated that the expression of TBP was the most stable at sulfide stress and in developmental process, while the expression of EF- 1- α was the most stable at sulfide stress and in various tissues. TBP and EF- 1- α were recommended as a suitable reference gene combination to accurately normalize the expression of target genes at sulfide stress; and EF- 1- α, TBP and TUB were considered as a potential reference gene combination for normalizing the expression of target genes in different tissues. No suitable gene combination was obtained among these five candidate genes for normalizing the expression of target genes for developmental process of U. unicinctus. Our results provided a valuable support for quantifying gene expression using RT-qPCR in U. unicinctus.

  2. Automated Real-Time Clearance Analyzer (ARCA), Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — The Automated Real-Time Clearance Analyzer (ARCA) addresses the future safety need for Real-Time System-Wide Safety Assurance (RSSA) in aviation and progressively...

  3. Specification and Automated Verification of Real-Time Behaviour

    DEFF Research Database (Denmark)

    Kristensen, C.H.; Andersen, J.H.; Skou, A.

    1995-01-01

    In this paper we sketch a method for specification and automatic verification of real-time software properties.......In this paper we sketch a method for specification and automatic verification of real-time software properties....

  4. Specification and Automated Verification of Real-Time Behaviour

    DEFF Research Database (Denmark)

    Andersen, J.H.; Kristensen, C.H.; Skou, A.

    1996-01-01

    In this paper we sketch a method for specification and automatic verification of real-time software properties.......In this paper we sketch a method for specification and automatic verification of real-time software properties....

  5. Internet-accessible real-time weather information system

    Digital Repository Service at National Institute of Oceanography (India)

    Desai, R.G.P.; Joseph, A.; Desa, E.; Mehra, P.; Desa, E.; Gouveia, A.D.

    An internet-accessible real-time weather information system has been developed. This system provides real-time accessibility to weather information from a multitude of spatially distributed weather stations. The Internet connectivity also offers...

  6. An integrated technique for developing real-time systems

    NARCIS (Netherlands)

    Hooman, J.J.M.; Vain, J.

    1995-01-01

    The integration of conceptual modeling techniques, formal specification, and compositional verification is considered for real time systems within the knowledge engineering context. We define constructive transformations from a conceptual meta model to a real time specification language and give

  7. Virtual timers in hierarchical real-time systems

    NARCIS (Netherlands)

    Heuvel, van den M.M.H.P.; Holenderski, M.J.; Cools, W.A.; Bril, R.J.; Lukkien, J.J.; Zhu, D.

    2009-01-01

    Hierarchical scheduling frameworks (HSFs) provide means for composing complex real-time systems from welldefined subsystems. This paper describes an approach to provide hierarchically scheduled real-time applications with virtual event timers, motivated by the need for integrating priority

  8. Spying on real-time computers to improve performance

    International Nuclear Information System (INIS)

    Taff, L.M.

    1975-01-01

    The sampled program-counter histogram, an established technique for shortening the execution times of programs, is described for a real-time computer. The use of a real-time clock allows particularly easy implementation. (Auth.)

  9. Real-time subway information for improving transit ridership.

    Science.gov (United States)

    2016-08-01

    In recent years, the standardization of transit schedule information has yielded a dramatic increase in the accessibility of computerized transit schedules and given rise to real-time service schedules. Two such real-time service schedules are the Ge...

  10. Improving Timeliness in Real-Time Secure Database Systems

    National Research Council Canada - National Science Library

    Son, Sang H; David, Rasikan; Thuraisingham, Bhavani

    2006-01-01

    .... In addition to real-time requirements, security is usually required in many applications. Multilevel security requirements introduce a new dimension to transaction processing in real-time database systems...

  11. On Real-Time Systems Using Local Area Networks.

    Science.gov (United States)

    1987-07-01

    87-35 July, 1987 CS-TR-1892 On Real - Time Systems Using Local Area Networks*I VShem-Tov Levi Department of Computer Science Satish K. Tripathit...1892 On Real - Time Systems Using Local Area Networks* Shem-Tov Levi Department of Computer Science Satish K. Tripathit Department of Computer Science...constraints and the clock systems that feed the time to real - time systems . A model for real-time system based on LAN communication is presented in

  12. Energy efficient approach for transient fault recovery in real time ...

    African Journals Online (AJOL)

    DR OKE

    Keywords: DVS, Fault tolerance, Real Time System, Transient Fault. ... in which missing the deadline may cause a failure and soft real time system, ..... Pillai, P., Shin, K., Real-time dynamic voltage scaling for low-power embedded operating ...

  13. ClockWork: a Real-Time Feasibility Analysis Tool

    NARCIS (Netherlands)

    Jansen, P.G.; Hanssen, F.T.Y.; Mullender, Sape J.

    ClockWork shows that we can improve the flexibility and efficiency of real-time kernels. We do this by proposing methods for scheduling based on so-called Real-Time Transactions. ClockWork uses Real-Time Transactions which allow scheduling decisions to be taken by the system. A programmer does not

  14. The real-time price elasticity of electricity

    NARCIS (Netherlands)

    Lijesen, M.G.

    2007-01-01

    The real-time price elasticity of electricity contains important information on the demand response of consumers to the volatility of peak prices. Despite the importance, empirical estimates of the real-time elasticity are hardly available. This paper provides a quantification of the real-time

  15. Temporal Specification and Verification of Real-Time Systems.

    Science.gov (United States)

    1991-08-30

    of concrete real - time systems can be modeled adequately. Specification: We present two conservative extensions of temporal logic that allow for the...logic. We present both model-checking algorithms for the automatic verification of finite-state real - time systems and proof methods for the deductive verification of real - time systems .

  16. Real-time volumetric scintillation dosimetry

    International Nuclear Information System (INIS)

    Beddar, S

    2015-01-01

    The goal of this brief review is to review the current status of real-time 3D scintillation dosimetry and what has been done so far in this area. The basic concept is to use a large volume of a scintillator material (liquid or solid) to measure or image the dose distributions from external radiation therapy (RT) beams in three dimensions. In this configuration, the scintillator material fulfills the dual role of being the detector and the phantom material in which the measurements are being performed. In this case, dose perturbations caused by the introduction of a detector within a phantom will not be at issue. All the detector configurations that have been conceived to date used a Charge-Coupled Device (CCD) camera to measure the light produced within the scintillator. In order to accurately measure the scintillation light, one must correct for various optical artefacts that arise as the light propagates from the scintillating centers through the optical chain to the CCD chip. Quenching, defined in its simplest form as a nonlinear response to high-linear energy transfer (LET) charged particles, is one of the disadvantages when such systems are used to measure the absorbed dose from high-LET particles such protons. However, correction methods that restore the linear dose response through the whole proton range have been proven to be effective for both liquid and plastic scintillators. Volumetric scintillation dosimetry has the potential to provide fast, high-resolution and accurate 3D imaging of RT dose distributions. Further research is warranted to optimize the necessary image reconstruction methods and optical corrections needed to achieve its full potential

  17. Real time PV manufacturing diagnostic system

    Energy Technology Data Exchange (ETDEWEB)

    Kochergin, Vladimir [MicroXact Inc., Blacksburg, VA (United States); Crawford, Michael A. [MicroXact Inc., Blacksburg, VA (United States)

    2015-09-01

    The main obstacle Photovoltaic (PV) industry is facing at present is the higher cost of PV energy compared to that of fossil energy. While solar cell efficiencies continue to make incremental gains these improvements are so far insufficient to drive PV costs down to match that of fossil energy. Improved in-line diagnostics however, has the potential to significantly increase the productivity and reduce cost by improving the yield of the process. On this Phase I/Phase II SBIR project MicroXact developed and demonstrated at CIGS pilot manufacturing line a high-throughput in-line PV manufacturing diagnostic system, which was verified to provide fast and accurate data on the spatial uniformity of thickness, an composition of the thin films comprising the solar cell as the solar cell is processed reel-to-reel. In Phase II project MicroXact developed a stand-alone system prototype and demonstrated the following technical characteristics: 1) ability of real time defect/composition inconsistency detection over 60cm wide web at web speeds up to 3m/minute; 2) Better than 1mm spatial resolution on 60cm wide web; 3) an average better than 20nm spectral resolution resulting in more than sufficient sensitivity to composition imperfections (copper-rich and copper-poor regions were detected). The system was verified to be high vacuum compatible. Phase II results completely validated both technical and economic feasibility of the proposed concept. MicroXact’s solution is an enabling technique for in-line PV manufacturing diagnostics to increase the productivity of PV manufacturing lines and reduce the cost of solar energy, thus reducing the US dependency on foreign oil while simultaneously reducing emission of greenhouse gasses.

  18. Novel functions of prototype foamy virus Gag glycine- arginine-rich boxes in reverse transcription and particle morphogenesis.

    Science.gov (United States)

    Müllers, Erik; Uhlig, Tobias; Stirnnagel, Kristin; Fiebig, Uwe; Zentgraf, Hanswalter; Lindemann, Dirk

    2011-02-01

    Prototype foamy virus (PFV) Gag lacks the characteristic orthoretroviral Cys-His motifs that are essential for various steps of the orthoretroviral replication cycle, such as RNA packaging, reverse transcription, infectivity, integration, and viral assembly. Instead, it contains three glycine-arginine-rich boxes (GR boxes) in its C terminus that putatively represent a functional equivalent. We used a four-plasmid replication-deficient PFV vector system, with uncoupled RNA genome packaging and structural protein translation, to analyze the effects of deletion and various substitution mutations within each GR box on particle release, particle-associated protein composition, RNA packaging, DNA content, infectivity, particle morphology, and intracellular localization. The degree of viral particle release by all mutants was similar to that of the wild type. Only minimal effects on Pol encapsidation, exogenous reverse transcriptase (RT) activity, and genomic viral RNA packaging were observed. In contrast, particle-associated DNA content and infectivity were drastically reduced for all deletion mutants and were undetectable for all alanine substitution mutants. Furthermore, GR box I mutants had significant changes in particle morphology, and GR box II mutants lacked the typical nuclear localization pattern of PFV Gag. Finally, it could be shown that GR boxes I and III, but not GR box II, can functionally complement each other. It therefore appears that, similar to the orthoretroviral Cys-His motifs, the PFV Gag GR boxes are important for RNA encapsidation, genome reverse transcription, and virion infectivity as well as for particle morphogenesis.

  19. Mixed - mode Operating System for Real - time Performance

    OpenAIRE

    Hasan M. M.; Sultana S.; Foo C.K.

    2017-01-01

    The purpose of the mixed-mode system research is to handle devices with the accuracy of real-time systems and at the same time, having all the benefits and facilities of a matured Graphic User Interface(GUI)operating system which is typicallynon-real-time. This mixed-mode operating system comprising of a real-time portion and a non-real-time portion was studied and implemented to identify the feasibilities and performances in practical applications (in the context of scheduled the real-time e...

  20. Detection of single-copy functional genes in prokaryotic cells by two-pass TSA-FISH with polynucleotide probes.

    Science.gov (United States)

    Kawakami, Shuji; Hasegawa, Takuya; Imachi, Hiroyuki; Yamaguchi, Takashi; Harada, Hideki; Ohashi, Akiyoshi; Kubota, Kengo

    2012-02-01

    In situ detection of functional genes with single-cell resolution is currently of interest to microbiologists. Here, we developed a two-pass tyramide signal amplification (TSA)-fluorescence in situ hybridization (FISH) protocol with PCR-derived polynucleotide probes for the detection of single-copy genes in prokaryotic cells. The mcrA gene and the apsA gene in methanogens and sulfate-reducing bacteria, respectively, were targeted. The protocol showed bright fluorescence with a good signal-to-noise ratio and achieved a high efficiency of detection (>98%). The discrimination threshold was approximately 82-89% sequence identity. Microorganisms possessing the mcrA or apsA gene in anaerobic sludge samples were successfully detected by two-pass TSA-FISH with polynucleotide probes. The developed protocol is useful for identifying single microbial cells based on functional gene sequences. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. Insertion sequence typing of Mycobacterium tuberculosis: characterization of a widespread subtype with a single copy of IS6110.

    Science.gov (United States)

    Fomukong, N G; Tang, T H; al-Maamary, S; Ibrahim, W A; Ramayah, S; Yates, M; Zainuddin, Z F; Dale, J W

    1994-12-01

    DNA fingerprinting with the insertion sequence IS6110 (also known as IS986) has become established as a major tool for investigating the spread of tuberculosis. Most strains of Mycobacterium tuberculosis have multiple copies of IS6110, but a small minority carry a single copy only. We have examined selected strains from Malaysia, Tanzania and Oman, in comparison with M. bovis isolates and BCG strains carrying one or two copies of IS6110. The insertion sequence appears to be present in the same position in all these strains, which suggests that in these organisms the element is defective in transposition and that the loss of transposability may have occurred at an early stage in the evolution of the M. tuberculosis complex.

  2. Real Time Wide Area Radiation Surveillance System

    Science.gov (United States)

    Biafore, M.

    2012-04-01

    We present the REWARD project, financed within the FP7 programme, theme SEC-2011.1.5-1 (Development of detection capabilities of difficult to detect radioactive sources and nuclear materials - Capability Project). Within this project, we propose a novel mobile system for real time, wide area radiation surveillance. The system is based on the integration of new miniaturized solid-state radiation sensors: a CdZnTe detector for gamma radiation and a high efficiency neutron detector based on novel silicon technologies. The sensing unit will include a wireless communication interface to send the data remotely to a monitoring base station which also uses a GPS system to calculate the position of the tag. The system will also incorporate middleware and high level software to provide web-service interfaces for the exchange of information, and that will offer top level functionalities as management of users, mobile tags and environment data and alarms, database storage and management and a web-based graphical user interface. Effort will be spent to ensure that the software is modular and re-usable across as many architectural levels as possible. Finally, an expert system will continuously analyze the information from the radiation sensor and correlate it with historical data from the tag location in order to generate an alarm when an abnormal situation is detected. The system will be useful for many different scenarios, including such lost radioactive sources and radioactive contamination. It will be possible to deploy in emergency units and in general in any type of mobile or static equipment. The sensing units will be highly portable thanks to their low size and low energy consumption. The complete system will be scalable in terms of complexity and cost and will offer very high precision on both the measurement and the location of the radiation. The modularity and flexibility of the system will allow for a realistic introduction to the market. Authorities may start with a

  3. Real Time Seismic Loss Estimation in Italy

    Science.gov (United States)

    Goretti, A.; Sabetta, F.

    2009-04-01

    By more than 15 years the Seismic Risk Office is able to perform a real-time evaluation of the earthquake potential loss in any part of Italy. Once the epicentre and the magnitude of the earthquake are made available by the National Institute for Geophysiscs and Volca-nology, the model, based on the Italian Geographic Information Sys-tems, is able to evaluate the extent of the damaged area and the consequences on the built environment. In recent years the model has been significantly improved with new methodologies able to conditioning the uncertainties using observa-tions coming from the fields during the first days after the event. However it is reputed that the main challenges in loss analysis are related to the input data, more than to methodologies. Unlike the ur-ban scenario, where the missing data can be collected with enough accuracy, the country-wise analysis requires the use of existing data bases, often collected for other purposed than seismic scenario evaluation, and hence in some way lacking of completeness and homogeneity. Soil properties, building inventory and population dis-tribution are the main input data that are to be known in any site of the whole Italian territory. To this end the National Census on Popu-lation and Dwellings has provided information on the residential building types and the population that lives in that building types. The critical buildings, such as Hospital, Fire Brigade Stations, Schools, are not included in the inventory, since the national plan for seismic risk assessment of critical buildings is still under way. The choice of a proper soil motion parameter, its attenuation with distance and the building type fragility are important ingredients of the model as well. The presentation will focus on the above mentioned issues, highlight-ing the different data sets used and their accuracy, and comparing the model, input data and results when geographical areas with dif-ferent extent are considered: from the urban scenarios

  4. Instrumentation development for real time brainwave monitoring.

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Lawrence Frederick; Clough, Benjamin W.

    2005-12-01

    The human brain functions through a chemically-induced biological process which operates in a manner similar to electrical systems. The signal resulting from this biochemical process can actually be monitored and read using tools and having patterns similar to those found in electrical and electronics engineering. The primary signature of this electrical activity is the ''brain wave'', which looks remarkably similar to the output of many electrical systems. Likewise, the device currently used in medical arenas to read brain electrical activity is the electroencephalogram (EEG) which is synonymous with a multi-channel oscilloscope reading. Brain wave readings and recordings for medical purposes are traditionally taken in clinical settings such as hospitals, laboratories or diagnostic clinics. The signal is captured via externally applied scalp electrodes using semi-viscous gel to reduce impedance. The signal will be in the 10 to 100 microvolt range. In other instances, where surgeons are attempting to isolate particular types of minute brain signals, the electrodes may actually be temporarily implanted in the brain during a preliminary procedure. The current configurations of equipment required for EEGs involve large recording instruments, many electrodes, wires, and large amounts of hard disk space devoted to storing large files of brain wave data which are then eventually analyzed for patterns of concern. Advances in sensors, signal processing, data storage and microelectronics over the last decade would seem to have paved the way for the realization of devices capable of ''real time'' external monitoring, and possible assessment, of brain activity. A myriad of applications for such a capability are likewise presenting themselves, including the ability to assess brain functioning, level of functioning and malfunctioning. Our plan is to develop the sensors, signal processing, and portable instrumentation package which could

  5. Real-time Forensic Disaster Analysis

    Science.gov (United States)

    Wenzel, F.; Daniell, J.; Khazai, B.; Mühr, B.; Kunz-Plapp, T.; Markus, M.; Vervaeck, A.

    2012-04-01

    The Center for Disaster Management and Risk Reduction Technology (CEDIM, www.cedim.de) - an interdisciplinary research center founded by the German Research Centre for Geoscience (GFZ) and Karlsruhe Institute of Technology (KIT) - has embarked on a new style of disaster research known as Forensic Disaster Analysis. The notion has been coined by the Integrated Research on Disaster Risk initiative (IRDR, www.irdrinternational.org) launched by ICSU in 2010. It has been defined as an approach to studying natural disasters that aims at uncovering the root causes of disasters through in-depth investigations that go beyond the reconnaissance reports and case studies typically conducted after disasters. In adopting this comprehensive understanding of disasters CEDIM adds a real-time component to the assessment and evaluation process. By comprehensive we mean that most if not all relevant aspects of disasters are considered and jointly analysed. This includes the impact (human, economy, and infrastructure), comparisons with recent historic events, social vulnerability, reconstruction and long-term impacts on livelihood issues. The forensic disaster analysis research mode is thus best characterized as "event-based research" through systematic investigation of critical issues arising after a disaster across various inter-related areas. The forensic approach requires (a) availability of global data bases regarding previous earthquake losses, socio-economic parameters, building stock information, etc.; (b) leveraging platforms such as the EERI clearing house, relief-web, and the many sources of local and international sources where information is organized; and (c) rapid access to critical information (e.g., crowd sourcing techniques) to improve our understanding of the complex dynamics of disasters. The main scientific questions being addressed are: What are critical factors that control loss of life, of infrastructure, and for economy? What are the critical interactions

  6. Real-time {sup 90}Sr Counter

    Energy Technology Data Exchange (ETDEWEB)

    Kaneko, Naomi; Kawai, Hideyuki; Kodama, Satoshi; Kobayashi, Atsushi; Tabata, Makoto; Ito, Hiroshi [Graduate School of Science, Chiba University, Chiba, (Japan); Han, Soorim [Graduate School of Science, Chiba University, Chiba, (Japan); National Institute of Radiological Science, Chiba, (Japan)

    2015-07-01

    Radioisotopes have been emitted around Japan due to a nuclear accident at the Fukushima Daiichi nuclear power station in March 2011. A problem is the contaminated water including the atomic nucleus which relatively has a long half- life time and soluble such as {sup 90}Sr, {sup 137}Cs. Internal exposures by {sup 90}Sr are more dangerous than {sup 137}Cs's because Sr has effective half-life time of 18 years and property of accumulation in a born. We have developed real-time {sup 90}Sr counter which is sensitive beta-ray of maximum kinematic energy of 2.28 MeV from {sup 90}Sr and insensitive of beta-ray of maximum kinematic energy of 1.17 MeV and gamma-ray from {sup 90}Sr by Cherenkov detection. This counter composes of Cerenkov counter, trigger scintillation counter and veto counter. Silica aerogel for Cherenkov counter can obtain refractive index between 1.017 and 1.049 easily. And wavelength shifting fiber (WLSF) is used as a light guide for extending effective area and producing lower cost. A mechanism of the identification of {sup 90}Sr is explained in following. In case of {sup 90}Sr, when the trigger counter reacts on the beta-ray from {sup 90}Sr, aerogel emits the Cherenkov light and WLSF reacts and read the Cherenkov light. On the other hand, in case of {sup 137}Cs, the trigger counter reacts on the beta-ray, aerogel stops the beta- ray and Cherenkov light is not emitted. Therefore, aerogel has a function as a radiator and shielding material. the gamma-ray is not reacted on the lower density detector. Cosmic rays would be also reacted by the veto counter. A prototype counter whose the effective area is 30 cm x 10 cm was obtained (2.0±1.2){sup 3} of mis-identification as {sup 137}Cs/{sup 90}Sr. Detection limit in the surface contamination inspection depends on measurement time and effective area mainly. The sensitivity of wide range, 10{sup -2} - 10{sup 4} Bq/cm{sup 2}, is obtained by adjustment of detection level in circuit of this counter. A lower

  7. Mixed - mode Operating System for Real - time Performance

    Directory of Open Access Journals (Sweden)

    Hasan M. M.

    2017-11-01

    Full Text Available The purpose of the mixed-mode system research is to handle devices with the accuracy of real-time systems and at the same time, having all the benefits and facilities of a matured Graphic User Interface(GUIoperating system which is typicallynon-real-time. This mixed-mode operating system comprising of a real-time portion and a non-real-time portion was studied and implemented to identify the feasibilities and performances in practical applications (in the context of scheduled the real-time events. In this research an i8751 microcontroller-based hardware was used to measure the performance of the system in real-time-only as well as non-real-time-only configurations. The real-time portion is an 486DX-40 IBM PC system running under DOS-based real-time kernel and the non-real-time portion is a Pentium IIIbased system running under Windows NT. It was found that mixed-mode systems performed as good as a typical real-time system and in fact, gave many additional benefits such as simplified/modular programming and load tolerance.

  8. Real-time data access layer for MDSplus

    International Nuclear Information System (INIS)

    Manduchi, G.; Luchetta, A.; Taliercio, C.; Fredian, T.; Stillerman, J.

    2008-01-01

    Recent extensions to MDSplus allow data handling in long discharges and provide a real-time data access and communication layer. The real-time data access layer is an additional component of MDSplus: it is possible to use the traditional MDSplus API during normal operation, and to select a subset of data items to be used in real time. Real-time notification is provided by a communication layer using a publish-subscribe pattern. The notification covers processes sharing the same data items even running on different machines, thus allowing the implementation of distributed control systems. The real-time data access layer has been developed for Windows, Linux, and VxWorks; it is currently being ported to Linux RTAI. In order to quantify the fingerprint of the presented system, the performance of the real-time access layer approach is compared with that of an ad hoc, manually optimized program in a sample real-time application

  9. Integration of MDSplus in real-time systems

    International Nuclear Information System (INIS)

    Luchetta, A.; Manduchi, G.; Taliercio, C.

    2006-01-01

    RFX-mod makes extensive usage of real-time systems for feedback control and uses MDSplus to interface them to the main Data Acquisition system. For this purpose, the core of MDSplus has been ported to VxWorks, the operating system used for real-time control in RFX. Using this approach, it is possible to integrate real-time systems, but MDSplus is used only for non-real-time tasks, i.e. those tasks which are executed before and after the pulse and whose performance does not affect the system time constraints. More extensive use of MDSplus in real-time systems is foreseen, and a real-time layer for MDSplus is under development, which will provide access to memory-mapped pulse files, shared by the tasks running on the same CPU. Real-time communication will also be integrated in the MDSplus core to provide support for distributed memory-mapped pulse files

  10. Rapid and sensitive detection of novel avian-origin influenza A (H7N9 virus by reverse transcription loop-mediated isothermal amplification combined with a lateral-flow device.

    Directory of Open Access Journals (Sweden)

    Yiyue Ge

    Full Text Available A severe disease in humans caused by a novel avian-origin influenza A (H7N9 virus emerged in China recently, which has caused at least 128 cases and 26 deaths. Rapid detection of the novel H7N9 virus is urgently needed to differentiate the disease from other infections, and to facilitate infection control as well as epidemiologic investigations. In this study, a reverse transcription loop-mediated isothermal amplification combined with a lateral flow device (RT-LAMP-LFD assay to rapidly detect H7N9 virus was developed and evaluated. The RT-LAMP primers were designed to target the haemagglutinin (HA and neuraminidase (NA genes of H7N9 virus. Results of 10-fold dilution series assays showed that analysis of RT-LAMP products by the LFD method was as sensitive as real-time turbidity detection, and that the analytic sensitivities of the HA and NA RT-LAMP assays were both 10 copies of synthetic RNA. Furthermore, both the assays showed 100% clinical specificity for identification of H7N9 virus. The performance characteristics of the RT-LAMP-LFD assay were evaluated with 80 clinical specimens collected from suspected H7N9 patients. The NA RT-LAMP-LFD assay was more sensitive than real time RT-PCR assay. Compared with a combination of virus culture and real-time RT-PCR, the sensitivity, specificity, positive predictive value, and negative predictive value of the RT-LAMP-LFD assay were all 100%. Overall, The RT-LAMP-LFD assay established in this study can be used as a reliable method for early diagnosis of the avian-origin influenza A (H7N9 virus infection.

  11. Characterization of the gut microbiota of Papua New Guineans using reverse transcription quantitative PCR.

    Directory of Open Access Journals (Sweden)

    Andrew R Greenhill

    Full Text Available There has been considerable interest in composition of gut microbiota in recent years, leading to a better understanding of the role the gut microbiota plays in health and disease. Most studies have been limited in their geographical and socioeconomic diversity to high-income settings, and have been conducted using small sample sizes. To date, few analyses have been conducted in low-income settings, where a better understanding of the gut microbiome could lead to the greatest return in terms of health benefits. Here, we have used quantitative real-time polymerase chain reaction targeting dominant and sub-dominant groups of microorganisms associated with human gut microbiome in 115 people living a subsistence lifestyle in rural areas of Papua New Guinea. Quantification of Clostridium coccoides group, C. leptum subgroup, C. perfringens, Bacteroides fragilis group, Bifidobacterium, Atopobium cluster, Prevotella, Enterobacteriaceae, Enterococcus, Staphylococcus, and Lactobacillus spp. was conducted. Principle coordinates analysis (PCoA revealed two dimensions with Prevotella, clostridia, Atopobium, Enterobacteriaceae, Enterococcus and Staphylococcus grouping in one dimension, while B. fragilis, Bifidobacterium and Lactobacillus grouping in the second dimension. Highland people had higher numbers of most groups of bacteria detected, and this is likely a key factor for the differences revealed by PCoA between highland and lowland study participants. Age and sex were not major determinants in microbial population composition. The study demonstrates a gut microbial composition with some similarities to those observed in other low-income settings where traditional diets are consumed, which have previously been suggested to favor energy extraction from a carbohydrate rich diet.

  12. Characterization of the gut microbiota of Papua New Guineans using reverse transcription quantitative PCR.

    Science.gov (United States)

    Greenhill, Andrew R; Tsuji, Hirokazu; Ogata, Kiyohito; Natsuhara, Kazumi; Morita, Ayako; Soli, Kevin; Larkins, Jo-Ann; Tadokoro, Kiyoshi; Odani, Shingo; Baba, Jun; Naito, Yuichi; Tomitsuka, Eriko; Nomoto, Koji; Siba, Peter M; Horwood, Paul F; Umezaki, Masahiro

    2015-01-01

    There has been considerable interest in composition of gut microbiota in recent years, leading to a better understanding of the role the gut microbiota plays in health and disease. Most studies have been limited in their geographical and socioeconomic diversity to high-income settings, and have been conducted using small sample sizes. To date, few analyses have been conducted in low-income settings, where a better understanding of the gut microbiome could lead to the greatest return in terms of health benefits. Here, we have used quantitative real-time polymerase chain reaction targeting dominant and sub-dominant groups of microorganisms associated with human gut microbiome in 115 people living a subsistence lifestyle in rural areas of Papua New Guinea. Quantification of Clostridium coccoides group, C. leptum subgroup, C. perfringens, Bacteroides fragilis group, Bifidobacterium, Atopobium cluster, Prevotella, Enterobacteriaceae, Enterococcus, Staphylococcus, and Lactobacillus spp. was conducted. Principle coordinates analysis (PCoA) revealed two dimensions with Prevotella, clostridia, Atopobium, Enterobacteriaceae, Enterococcus and Staphylococcus grouping in one dimension, while B. fragilis, Bifidobacterium and Lactobacillus grouping in the second dimension. Highland people had higher numbers of most groups of bacteria detected, and this is likely a key factor for the differences revealed by PCoA between highland and lowland study participants. Age and sex were not major determinants in microbial population composition. The study demonstrates a gut microbial composition with some similarities to those observed in other low-income settings where traditional diets are consumed, which have previously been suggested to favor energy extraction from a carbohydrate rich diet.

  13. A Short Interspersed Nuclear Element (SINE)-Based Real-Time PCR Approach to Detect and Quantify Porcine Component in Meat Products.

    Science.gov (United States)

    Zhang, Chi; Fang, Xin; Qiu, Haopu; Li, Ning

    2015-01-01

    Real-time PCR amplification of mitochondria gene could not be used for DNA quantification, and that of single copy DNA did not allow an ideal sensitivity. Moreover, cross-reactions among similar species were commonly observed in the published methods amplifying repetitive sequence, which hindered their further application. The purpose of this study was to establish a short interspersed nuclear element (SINE)-based real-time PCR approach having high specificity for species detection that could be used in DNA quantification. After massive screening of candidate Sus scrofa SINEs, one optimal combination of primers and probe was selected, which had no cross-reaction with other common meat species. LOD of the method was 44 fg DNA/reaction. Further, quantification tests showed this approach was practical in DNA estimation without tissue variance. Thus, this study provided a new tool for qualitative detection of porcine component, which could be promising in the QC of meat products.

  14. Real-Time Wireless Data Acquisition System

    Science.gov (United States)

    Valencia, Emilio J.; Perotti, Jose; Lucena, Angel; Mata, Carlos

    2007-01-01

    Current and future aerospace requirements demand the creation of a new breed of sensing devices, with emphasis on reduced weight, power consumption, and physical size. This new generation of sensors must possess a high degree of intelligence to provide critical data efficiently and in real-time. Intelligence will include self-calibration, self-health assessment, and pre-processing of raw data at the sensor level. Most of these features are already incorporated in the Wireless Sensors Network (SensorNet(TradeMark)), developed by the Instrumentation Group at Kennedy Space Center (KSC). A system based on the SensorNet(TradeMark) architecture consists of data collection point(s) called Central Stations (CS) and intelligent sensors called Remote Stations (RS) where one or more CSs can be accommodated depending on the specific application. The CS's major function is to establish communications with the Remote Stations and to poll each RS for data and health information. The CS also collects, stores and distributes these data to the appropriate systems requiring the information. The system has the ability to perform point-to-point, multi-point and relay mode communications with an autonomous self-diagnosis of each communications link. Upon detection of a communication failure, the system automatically reconfigures to establish new communication paths. These communication paths are automatically and autonomously selected as the best paths by the system based on the existing operating environment. The data acquisition system currently under development at KSC consists of the SensorNet(TradeMark) wireless sensors as the remote stations and the central station called the Radio Frequency Health Node (RFHN). The RFF1N is the central station which remotely communicates with the SensorNet(TradeMark) sensors to control them and to receive data. The system's salient feature is the ability to provide deterministic sensor data with accurate time stamps for both time critical and non

  15. Real time measurement of air radioactivity

    International Nuclear Information System (INIS)

    Galeriu, D.; Craciunescu, T.; Teles, S.

    1998-01-01

    commands to the logger according to the command protocol. - ODAS reads the configuration from logger. At initialization the software sends to the logger the desired configuration which is overlooked in the following operations. It calculates the averages of readings over 10 minute and other parameters. Following that the software records the data into a file (one line of data at every 10 min). The interrogation of logger and data collection is repeated at every 10 minute. - After every file recording, ODAS sends a code signal to the workstation. Transmission of data to RODOS workstation is done periodically (every 10 min) or when requested by users. Advantages of our Local Meteorological and Radiological Monitoring System are the following: - it is automate, able to acquire, calculate and transmit data in non-stop mode; - it is a real-time and on-line system; - it is easy to upgrade due to the fact that it was developed as modules working independently. (authors)

  16. Hardware locks for a real-time Java chip multiprocessor

    DEFF Research Database (Denmark)

    Strøm, Torur Biskopstø; Puffitsch, Wolfgang; Schoeberl, Martin

    2016-01-01

    A software locking mechanism commonly protects shared resources for multithreaded applications. This mechanism can, especially in chip-multiprocessor systems, result in a large synchronization overhead. For real-time systems in particular, this overhead increases the worst-case execution time....... This improvement can allow a larger number of real-time tasks to be reliably scheduled on a multiprocessor real-time platform....

  17. 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.

  18. A Programmable Microkernel for Real-Time Systems

    Science.gov (United States)

    2003-06-01

    A Programmable Microkernel for Real - Time Systems Christoph M. Kirsch Thomas A. Henzinger Marco A.A. Sanvido Report No. UCB/CSD-3-1250 June 2003...TITLE AND SUBTITLE A Programmable Microkernel for Real - Time Systems 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S...THIS PAGE unclassified Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18 A Programmable Microkernel for Real - Time Systems ∗ Christoph M

  19. Design Specifications for Adaptive Real-Time Systems

    Science.gov (United States)

    1991-12-01

    TICfl \\ E CT E Design Specifications for JAN’\\ 1992 Adaptive Real - Time Systems fl Randall W. Lichota U, Alice H. Muntz - December 1991 \\ \\\\/ 0 / r...268-2056 Technical Report CMU/SEI-91-TR-20 ESD-91-TR-20 December 1991 Design Specifications for Adaptive Real - Time Systems Randall W. Lichota Hughes...Design Specifications for Adaptive Real - Time Systems Abstract: The design specification method described in this report treats a software

  20. Temporal Proof Methodologies for Real-Time Systems,

    Science.gov (United States)

    1990-09-01

    real time systems that communicate either through shared variables or by message passing and real time issues such as time-outs, process priorities (interrupts) and process scheduling. The authors exhibit two styles for the specification of real - time systems . While the first approach uses bounded versions of temporal operators the second approach allows explicit references to time through a special clock variable. Corresponding to two styles of specification the authors present and compare two fundamentally different proof

  1. Proceedings of the Real-Time Systems Engineering Workshop

    Science.gov (United States)

    2001-08-01

    real - time systems engineering. The workshop was held as part of the SEI Symposium in...Washington, DC, during September 2000. The objective of the workshop was to identify key issues and obtain feedback from attendees concerning real - time systems engineering...and interoperability. This report summarizes the workshop in terms of foundation, management, and technical topics, and it contains a discussion related to developing a community of interest for real - time systems

  2. Static Schedulers for Embedded Real-Time Systems

    Science.gov (United States)

    1989-12-01

    Because of the need for having efficient scheduling algorithms in large scale real time systems , software engineers put a lot of effort on developing...provide static schedulers for he Embedded Real Time Systems with single processor using Ada programming language. The independent nonpreemptable...support the Computer Aided Rapid Prototyping for Embedded Real Time Systems so that we determine whether the system, as designed, meets the required

  3. PERTS: A Prototyping Environment for Real-Time Systems

    Science.gov (United States)

    Liu, Jane W. S.; Lin, Kwei-Jay; Liu, C. L.

    1993-01-01

    PERTS is a prototyping environment for real-time systems. It is being built incrementally and will contain basic building blocks of operating systems for time-critical applications, tools, and performance models for the analysis, evaluation and measurement of real-time systems and a simulation/emulation environment. It is designed to support the use and evaluation of new design approaches, experimentations with alternative system building blocks, and the analysis and performance profiling of prototype real-time systems.

  4. Design Recovery Technology for Real-Time Systems.

    Science.gov (United States)

    1995-10-01

    RL-TR-95-208 Final Technical Report October 1995 DESIGN RECOVERY TECHNOLOGY FOR REAL TIME SYSTEMS The MITRE Corporation Lester J. Holtzblatt...92 - Jan 95 4. TTTLE AND SUBTITLE DESIGN RECOVERY TECHNOLOGY FOR REAL - TIME SYSTEMS 6. AUTHOR(S) Lester J. Holtzblatt, Richard Piazza, and Susan...behavior of real - time systems in general, our initial efforts have centered on recovering this information from one system in particular, the Modular

  5. Simultaneous detection of four garlic viruses by multiplex reverse transcription PCR and their distribution in Indian garlic accessions.

    Science.gov (United States)

    Majumder, S; Baranwal, V K

    2014-06-01

    Indian garlic is infected with Onion yellow dwarf virus (OYDV), Shallot latent virus (SLV), Garlic common latent virus (GarCLV) and allexiviruses. Identity and distribution of garlic viruses in various garlic accessions from different geographical regions of India were investigated. OYDV and allexiviruses were observed in all the garlic accessions, while SLV and GarCLV were observed only in a few accessions. A multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection and identification of OYDV, SLV, GarCLV and Allexivirus infecting garlic accessions in India. This multiplex protocol standardized in this study will be useful in indexing of garlic viruses and production of virus free seed material. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Detection of foot-and-mouth disease virus rna by reverse transcription loop-mediated isothermal amplification

    Directory of Open Access Journals (Sweden)

    Chen Hao-tai

    2011-11-01

    Full Text Available Abstract A reverse transcription loop-mediated isothermal amplification (RT-LAMP assay was developed for foot-and-mouth disease virus (FMDV RNA. The amplification was able to finish in 45 min under isothermal condition at 64°C by employing a set of four primers targeting FMDV 2B. The assay showed higher sensitivity than RT-PCR. No cross reactivity was observed from other RNA viruses including classical swine fever virus, swine vesicular disease, porcine reproductive and respiratory syndrome virus, Japanese encephalitis virus. Furthermore, the assay correctly detected 84 FMDV positive samples but not 65 FMDV negative specimens. The result indicated the potential usefulness of the technique as a simple and rapid procedure for the detection of FMDV infection.

  7. Simultaneous detection of enteropathogenic viruses in buffalos faeces using multiplex reverse transcription-polymerase chain reaction (mRT-PCR

    Directory of Open Access Journals (Sweden)

    U. Pagnini

    2010-02-01

    Full Text Available A multiplex reverse transcription- polymerase chain reaction (mRT-PCR assay that detects Bovine Viral Diarrhoea Virus, Bovine Coronavirus, and Group A Rotaviruses in infected cell-culture fluids and clinical faecal samples is described. One hundred twenty faecal samples from buffalo calves with acute gastroenteritis were tested. The mRT-PCR was validated against simplex RT-PCR with published primers for Pestivirus, Coronavirus and Rotavirus. The multiplex RT-PCR was equally sensitive and specific in detecting viral infections compared with simplex RT-PCR. The mRT-PCR readily identified viruses by discriminating the size of their amplified gene products. This mRT-PCR may be a sensitive and rapid assay for surveillance of buffalo enteric viruses in field specimens. This novel multiplex RT-PCR is an attractive technique for the rapid, specific, and cost-effective laboratory diagnosis of acute gastroenteritis.

  8. Mixed-mode Operating System for Real-time Performance

    OpenAIRE

    M.M. Hasan; S. Sultana; C.K. Foo

    2017-01-01

    The purpose of the mixed-mode system research is to handle devices with the accuracy of real-time systems and at the same time, having all the benefits and facilities of a matured Graphic User Interface (GUI) operating system which is typically nonreal-time. This mixed-mode operating system comprising of a real-time portion and a non-real-time portion was studied and implemented to identify the feasibilities and performances in practical applications (in the context of scheduled the real-time...

  9. Timeliness and Predictability in Real-Time Database Systems

    National Research Council Canada - National Science Library

    Son, Sang H

    1998-01-01

    The confluence of computers, communications, and databases is quickly creating a globally distributed database where many applications require real time access to both temporally accurate and multimedia data...

  10. Verifying real-time systems against scenario-based requirements

    DEFF Research Database (Denmark)

    Larsen, Kim Guldstrand; Li, Shuhao; Nielsen, Brian

    2009-01-01

    We propose an approach to automatic verification of real-time systems against scenario-based requirements. A real-time system is modeled as a network of Timed Automata (TA), and a scenario-based requirement is specified as a Live Sequence Chart (LSC). We define a trace-based semantics for a kernel...... subset of the LSC language. By equivalently translating an LSC chart into an observer TA and then non-intrusively composing this observer with the original system model, the problem of verifying a real-time system against a scenario-based requirement reduces to a classical real-time model checking...

  11. Building Real-Time Collaborative Applications with a Federated Architecture

    Directory of Open Access Journals (Sweden)

    Pablo Ojanguren-Menendez

    2015-12-01

    Full Text Available Real-time collaboration is being offered by multiple libraries and APIs (Google Drive Real-time API, Microsoft Real-Time Communications API, TogetherJS, ShareJS, rapidly becoming a mainstream option for webservices developers. However, they are offered as centralised services running in a single server, regardless if they are free/open source or proprietary software. After re-engineering Apache Wave (former Google Wave, we can now provide the first decentralised and federated free/open source alternative. The new API allows to develop new real-time collaborative web applications in both JavaScript and Java environments.

  12. External Quality Assessment for the Detection of Measles Virus by Reverse Transcription-PCR Using Armored RNA.

    Directory of Open Access Journals (Sweden)

    Dong Zhang

    Full Text Available In recent years, nucleic acid tests for detection of measles virus RNA have been widely applied in laboratories belonging to the measles surveillance system of China. An external quality assessment program was established by the National Center for Clinical Laboratories to evaluate the performance of nucleic acid tests for measles virus. The external quality assessment panel, which consisted of 10 specimens, was prepared using armored RNAs, complex of noninfectious MS2 bacteriophage coat proteins encapsulated RNA of measles virus, as measles virus surrogate controls. Conserved sequences amplified from a circulating measles virus strain or from a vaccine strain were encapsulated into these armored RNAs. Forty-one participating laboratories from 15 provinces, municipalities, or autonomous regions that currently conduct molecular detection of measles virus enrolled in the external quality assessment program, including 40 measles surveillance system laboratories and one diagnostic reagent manufacturer. Forty laboratories used commercial reverse transcription-quantitative PCR kits, with only one laboratory applying a conventional PCR method developed in-house. The results indicated that most of the participants (38/41, 92.7% were able to accurately detect the panel with 100% sensitivity and 100% specificity. Although a wide range of commercially available kits for nucleic acid extraction and reverse transcription polymerase chain reaction were used by the participants, only two false-negative results and one false-positive result were generated; these were generated by three separate laboratories. Both false-negative results were obtained with tests performed on specimens with the lowest concentration (1.2 × 104 genomic equivalents/mL. In addition, all 18 participants from Beijing achieved 100% sensitivity and 100% specificity. Overall, we conclude that the majority of the laboratories evaluated have reliable diagnostic capacities for the detection

  13. Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus

    International Nuclear Information System (INIS)

    Huang, S-H; Tsai, M-H; Lin, C-W; Yang, T-C; Chuang, P-H; Tsai, I-S; Lu, H-C; Wan Lei; Lin, Y-J; Lai, C-H

    2008-01-01

    Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples

  14. Evaluation of a multiplex real-time PCR assay for the detection of respiratory viruses in clinical specimens.

    Science.gov (United States)

    Rheem, Insoo; Park, Joowon; Kim, Tae-Hyun; Kim, Jong Wan

    2012-11-01

    In this study, we evaluated the analytical performance and clinical potential of a one-step multiplex real-time PCR assay for the simultaneous detection of 14 types of respiratory viruses using the AdvanSure RV real-time PCR Kit (LG Life Sciences, Korea). Three hundred and twenty clinical specimens were tested with the AdvanSure RV real-time PCR Kit and conventional multiplex reverse transcription (RT)-PCR assay. The assay results were analyzed and the one-step AdvanSure RV real-time PCR Kit was compared with the conventional multiplex RT-PCR assay with respect to the sensitivity and specificity of the detection of respiratory viruses. The limit of detection (LOD) was 1.31 plaque-forming units (PFU)/mL for human rhinoviruses (hRVs), 4.93 PFU/mL for human coronavirus HCoV-229E/NL63, 2.67 PFU/mL for human coronavirus HCoV-OC43, 18.20 PFU/mL for parainfluenza virus 1 (PIV)-1, 24.57 PFU/mL for PIV-2, 1.73 PFU/mL for PIV-3, 1.79 PFU/mL for influenza virus group (Flu) A, 59.51 PFU/mL for FluB, 5.46 PFU/mL for human respiratory syncytial virus (hRSV)-A, 17.23 PFU/mL for hRSV-B, 9.99 PFU/mL for human adenovirus (ADVs). The cross-reactivity test for this assay against 23 types of non-respiratory viruses showed negative results for all viruses tested. The agreement between the one-step AdvanSure multiplex real-time PCR assay and the conventional multiplex RT-PCR assay was 98%. The one-step AdvanSure RV multiplex real-time PCR assay is a simple assay with high potential for specific, rapid and sensitive laboratory diagnosis of respiratory viruses compared to conventional multiplex RT-PCR.

  15. GNSS global real-time augmentation positioning: Real-time precise satellite clock estimation, prototype system construction and performance analysis

    Science.gov (United States)

    Chen, Liang; Zhao, Qile; Hu, Zhigang; Jiang, Xinyuan; Geng, Changjiang; Ge, Maorong; Shi, Chuang

    2018-01-01

    Lots of ambiguities in un-differenced (UD) model lead to lower calculation efficiency, which isn't appropriate for the high-frequency real-time GNSS clock estimation, like 1 Hz. Mixed differenced model fusing UD pseudo-range and epoch-differenced (ED) phase observations has been introduced into real-time clock estimation. In this contribution, we extend the mixed differenced model for realizing multi-GNSS real-time clock high-frequency updating and a rigorous comparison and analysis on same conditions are performed to achieve the best real-time clock estimation performance taking the efficiency, accuracy, consistency and reliability into consideration. Based on the multi-GNSS real-time data streams provided by multi-GNSS Experiment (MGEX) and Wuhan University, GPS + BeiDou + Galileo global real-time augmentation positioning prototype system is designed and constructed, including real-time precise orbit determination, real-time precise clock estimation, real-time Precise Point Positioning (RT-PPP) and real-time Standard Point Positioning (RT-SPP). The statistical analysis of the 6 h-predicted real-time orbits shows that the root mean square (RMS) in radial direction is about 1-5 cm for GPS, Beidou MEO and Galileo satellites and about 10 cm for Beidou GEO and IGSO satellites. Using the mixed differenced estimation model, the prototype system can realize high-efficient real-time satellite absolute clock estimation with no constant clock-bias and can be used for high-frequency augmentation message updating (such as 1 Hz). The real-time augmentation message signal-in-space ranging error (SISRE), a comprehensive accuracy of orbit and clock and effecting the users' actual positioning performance, is introduced to evaluate and analyze the performance of GPS + BeiDou + Galileo global real-time augmentation positioning system. The statistical analysis of real-time augmentation message SISRE is about 4-7 cm for GPS, whlile 10 cm for Beidou IGSO/MEO, Galileo and about 30 cm

  16. Analysis of T-DNA/Host-Plant DNA Junction Sequences in Single-Copy Transgenic Barley Lines

    Directory of Open Access Journals (Sweden)

    Joanne G. Bartlett

    2014-01-01

    Full Text Available Sequencing across the junction between an integrated transfer DNA (T-DNA and a host plant genome provides two important pieces of information. The junctions themselves provide information regarding the proportion of T-DNA which has integrated into the host plant genome, whilst the transgene flanking sequences can be used to study the local genetic environment of the integrated transgene. In addition, this information is important in the safety assessment of GM crops and essential for GM traceability. In this study, a detailed analysis was carried out on the right-border T-DNA junction sequences of single-copy independent transgenic barley lines. T-DNA truncations at the right-border were found to be relatively common and affected 33.3% of the lines. In addition, 14.3% of lines had rearranged construct sequence after the right border break-point. An in depth analysis of the host-plant flanking sequences revealed that a significant proportion of the T-DNAs integrated into or close to known repetitive elements. However, this integration into repetitive DNA did not have a negative effect on transgene expression.

  17. Single-copy nuclear genes place haustorial Hydnoraceae within piperales and reveal a cretaceous origin of multiple parasitic angiosperm lineages.

    Directory of Open Access Journals (Sweden)

    Julia Naumann

    Full Text Available Extreme haustorial parasites have long captured the interest of naturalists and scientists with their greatly reduced and highly specialized morphology. Along with the reduction or loss of photosynthesis, the plastid genome often decays as photosynthetic genes are released from selective constraint. This makes it challenging to use traditional plastid genes for parasitic plant phylogenetics, and has driven the search for alternative phylogenetic and molecular evolutionary markers. Thus, evolutionary studies, such as molecular clock-based age estimates, are not yet available for all parasitic lineages. In the present study, we extracted 14 nuclear single copy genes (nSCG from Illumina transcriptome data from one of the "strangest plants in the world", Hydnora visseri (Hydnoraceae. A ~15,000 character molecular dataset, based on all three genomic compartments, shows the utility of nSCG for reconstructing phylogenetic relationships in parasitic lineages. A relaxed molecular clock approach with the same multi-locus dataset, revealed an ancient age of ~91 MYA for Hydnoraceae. We then estimated the stem ages of all independently originated parasitic angiosperm lineages using a published dataset, which also revealed a Cretaceous origin for Balanophoraceae, Cynomoriaceae and Apodanthaceae. With the exception of Santalales, older parasite lineages tend to be more specialized with respect to trophic level and have lower species diversity. We thus propose the "temporal specialization hypothesis" (TSH implementing multiple independent specialization processes over time during parasitic angiosperm evolution.

  18. Interspersion of highly repetitive DNA with single copy DNA in the genome of the red crab, Geryon quinquedens

    Energy Technology Data Exchange (ETDEWEB)

    Christie, N.T. (Univ. of Tennessee, Oak Ridge); Skinner, D.M.

    1979-02-01

    Kinetic analysis of the reassociation of 420 nucleotide (NT) long fragments has shown that essentially all of the repetitive sequences of the DNA of the red crab Geryon quinquedens are highly repetitive. There are negligible amounts of low and intermediate repetitive DNAs. Though atypical of most eukaryotes, this pattern has been observed in al other brachyurans (true crabs) studied. The major repetitive component is subdivided into short runs of 300 NT and longer runs of greater than 1200 NT while the minor component has an average sequence length of 400 NT. Both components reassociate at rates commonly observed for satellite DNAs. Unique among eukaryotes the organization of the genome includes single copy DNA contiguous to short runs (300 NT) of both repetitive components. Although patent satellites are not present, subsets of the repetitive DNA have been isolated by either restriction endonuclease digestion or by centrifugation in Ag/sup +/ or Hg/sup 2 +//Cs/sub 2/SO/sub 4/ density gradients.

  19. Flexible Scheduling by Deadline Inheritance in Soft Real Time Kernels

    NARCIS (Netherlands)

    Jansen, P.G.; Wygerink, Emiel

    1996-01-01

    Current Hard Real Time (HRT) kernels have their timely behaviour guaranteed on the cost of a rather restrictive use of the available resources. This makes HRT scheduling techniques inadequate for use in Soft Real Time (SRT) environment where we can make a considerable profit by a better and more

  20. Real-Time Plasma Control Tools for Advanced Tokamak Operation

    International Nuclear Information System (INIS)

    Varandas, C. A. F.; Sousa, J.; Rodrigues, A. P.; Carvalho, B. B.; Fernandes, H.; Batista, A. J.; Cruz, N.; Combo, A.; Pereira, R. C.

    2006-01-01

    Real-time control will play an important role in the operation and scientific exploitation of the new generation fusion devices. This paper summarizes the real-time systems and diagnostics developed by the Portuguese Fusion Euratom Association based on digital signal processors and field programmable gate arrays

  1. Real-time software for the COMPASS tokamak plasma control

    International Nuclear Information System (INIS)

    Valcarcel, D.F.; Duarte, A.S.; Neto, A.; Carvalho, I.S.; Carvalho, B.B.; Fernandes, H.; Sousa, J.; Sartori, F.; Janky, F.; Cahyna, P.; Hron, M.; Panek, R.

    2010-01-01

    The COMPASS tokamak has started its operation recently in Prague and to meet the necessary operation parameters its real-time system, for data processing and control, must be designed for both flexibility and performance, allowing the easy integration of code from several developers and to guarantee the desired time cycle. For this purpose an Advanced Telecommunications Computing Architecture based real-time system has been deployed with a solution built on a multi-core x86 processor. It makes use of two software components: the BaseLib2 and the MARTe (Multithreaded Application Real-Time executor) real-time frameworks. The BaseLib2 framework is a generic real-time library with optimized objects for the implementation of real-time algorithms. This allowed to build a library of modules that process the acquired data and execute control algorithms. MARTe executes these modules in kernel space Real-Time Application Interface allowing to attain the required cycle time and a jitter of less than 1.5 μs. MARTe configuration and data storage are accomplished through a Java hardware client that connects to the FireSignal control and data acquisition software. This article details the implementation of the real-time system for the COMPASS tokamak, in particular the organization of the control code, the design and implementation of the communications with the actuators and how MARTe integrates with the FireSignal software.

  2. Mixed-mode Operating System for Real-time Performance

    Directory of Open Access Journals (Sweden)

    M.M. Hasan

    2017-11-01

    Full Text Available The purpose of the mixed-mode system research is to handle devices with the accuracy of real-time systems and at the same time, having all the benefits and facilities of a matured Graphic User Interface (GUI operating system which is typically nonreal-time. This mixed-mode operating system comprising of a real-time portion and a non-real-time portion was studied and implemented to identify the feasibilities and performances in practical applications (in the context of scheduled the real-time events. In this research an i8751 microcontroller-based hardware was used to measure the performance of the system in real-time-only as well as non-real-time-only configurations. The real-time portion is an 486DX-40 IBM PC system running under DOS-based realtime kernel and the non-real-time portion is a Pentium III based system running under Windows NT. It was found that mixed-mode systems performed as good as a typical realtime system and in fact, gave many additional benefits such as simplified/modular programming and load tolerance.

  3. An algorithm for learning real-time automata

    NARCIS (Netherlands)

    Verwer, S.E.; De Weerdt, M.M.; Witteveen, C.

    2007-01-01

    We describe an algorithm for learning simple timed automata, known as real-time automata. The transitions of real-time automata can have a temporal constraint on the time of occurrence of the current symbol relative to the previous symbol. The learning algorithm is similar to the redblue fringe

  4. Real-Time Adaptation of Influence Strategies in Online Selling

    NARCIS (Netherlands)

    Kaptein, M.C.; Parvinen, P.

    2014-01-01

    Real-time adjustments in online selling are becoming increasingly common. In this paper we describe a novel method of real-time adaptation, and introduce influence strategies as a useful level of analysis for personalization of online selling. The proposed method incorporates three perspectives on

  5. 76 FR 42536 - Real-Time System Management Information Program

    Science.gov (United States)

    2011-07-19

    ...-Time System Management Information Program AGENCY: Federal Highway Administration (FHWA), DOT. ACTION... Real-Time System Management Information Program and general information about current and planned... establishing requirements for the Real-Time System Management Information Program on November 8, 2010, at 75 FR...

  6. 75 FR 68418 - Real-Time System Management Information Program

    Science.gov (United States)

    2010-11-08

    ...-Time System Management Information Program AGENCY: Federal Highway Administration (FHWA), DOT. ACTION...) to establish a Real-Time System Management Information Program that provides, in all States, the... traveler information. The purposes of the Real-Time System Management Information Program are to: (1...

  7. Compilation and synthesis for real-time embedded controllers

    DEFF Research Database (Denmark)

    Fränzle, Martin; Müller-Olm, Markus

    1999-01-01

    This article provides an overview over two constructive approaches to provably correct hard real-time code generation where hard real-time code is generated from abstract requirements rather than verified against the timing requirements a posteriori. The first, more pragmatic approach is concerne......-time systems at a very high level of abstraction....

  8. Real-Time Engagement Area Development Program (READ-Pro)

    National Research Council Canada - National Science Library

    Burger, Joseph

    2002-01-01

    The Real Time Engagement Area Development Program (READ-Pro) is a PC-based prototype system which provides company-level commanders with real-time operational analysis tools to develop ENGAGEMENT AREAS(EA) for direct fire (DF) systems...

  9. Real-time centre detection of an OLED structure

    NARCIS (Netherlands)

    Pieters, R.S.; Jonker, P.P.; Nijmeijer, H.

    2009-01-01

    The research presented in this paper focuses on real-time image processing for visual servoing, i.e. the positioning of a x-y table by using a camera only instead of encoders. A camera image stream plus real-time image processing determines the position in the next iteration of the table controller.

  10. Real-Time Center Detection of an OLED Structure

    NARCIS (Netherlands)

    Pieters, R.S.; Jonker, P.P.; Nijmeijer, H.; Blanc-Talon, J.; Philips, W.; Popescu, D.; Scheunders, P.

    2009-01-01

    The research presented in this paper focuses on real-time image processing for visual servoing, i.e. the positioning of a x-y table by using a camera only instead of encoders. A camera image stream plus real-time image processing determines the position in the next iteration of the table controller.

  11. Real-Time PCR for Universal Phytoplasma Detection and Quantification

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Nyskjold, Henriette; Nicolaisen, Mogens

    2013-01-01

    Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs and in other...

  12. Real-Time Operating Systems for Multicore Embedded Systems

    OpenAIRE

    Tomiyama, Hiroyuki; Honda, Shinya; Takada, Hiroaki

    2008-01-01

    Multicore systems-on-chip have become popular inthe design of embedded systems in order to simultaneously achieve high performance and low power consumption. On the software side, real-time operating systems are necessary in orderto handle growing complexity of embedded software. This paper describes requirements, design principles and implementation techniques for real-time operating systems to be used inasymmetric multicore systems.

  13. Parametric Room Acoustic workflows with real-time acoustic simulation

    DEFF Research Database (Denmark)

    Parigi, Dario

    2017-01-01

    The paper investigates and assesses the opportunities that real-time acoustic simulation offer to engage in parametric acoustics workflow and to influence architectural designs from early design stages......The paper investigates and assesses the opportunities that real-time acoustic simulation offer to engage in parametric acoustics workflow and to influence architectural designs from early design stages...

  14. Process algebra with timing : real time and discrete time

    NARCIS (Netherlands)

    Baeten, J.C.M.; Middelburg, C.A.; Bergstra, J.A.; Ponse, A.J.; Smolka, S.A.

    2001-01-01

    We present real time and discrete time versions of ACP with absolute timing and relative timing. The starting-point is a new real time version with absolute timing, called ACPsat, featuring urgent actions and a delay operator. The discrete time versions are conservative extensions of the discrete

  15. Real-time software for the COMPASS tokamak plasma control

    Energy Technology Data Exchange (ETDEWEB)

    Valcarcel, D.F., E-mail: danielv@ipfn.ist.utl.p [Associacao EURATOM/IST, Instituto de Plasmas e Fusao Nuclear - Laboratorio Associado, Instituto Superior Tecnico, P-1049-001 Lisboa (Portugal); Duarte, A.S.; Neto, A.; Carvalho, I.S.; Carvalho, B.B.; Fernandes, H.; Sousa, J. [Associacao EURATOM/IST, Instituto de Plasmas e Fusao Nuclear - Laboratorio Associado, Instituto Superior Tecnico, P-1049-001 Lisboa (Portugal); Sartori, F. [Euratom-UKAEA, Culham Science Centre, Abingdon, OX14 3DB Oxon (United Kingdom); Janky, F.; Cahyna, P.; Hron, M.; Panek, R. [Institute of Plasma Physics AS CR, v.v.i., Association EURATOM/IPP.CR, Za Slovankou 3, 182 00 Prague (Czech Republic)

    2010-07-15

    The COMPASS tokamak has started its operation recently in Prague and to meet the necessary operation parameters its real-time system, for data processing and control, must be designed for both flexibility and performance, allowing the easy integration of code from several developers and to guarantee the desired time cycle. For this purpose an Advanced Telecommunications Computing Architecture based real-time system has been deployed with a solution built on a multi-core x86 processor. It makes use of two software components: the BaseLib2 and the MARTe (Multithreaded Application Real-Time executor) real-time frameworks. The BaseLib2 framework is a generic real-time library with optimized objects for the implementation of real-time algorithms. This allowed to build a library of modules that process the acquired data and execute control algorithms. MARTe executes these modules in kernel space Real-Time Application Interface allowing to attain the required cycle time and a jitter of less than 1.5 {mu}s. MARTe configuration and data storage are accomplished through a Java hardware client that connects to the FireSignal control and data acquisition software. This article details the implementation of the real-time system for the COMPASS tokamak, in particular the organization of the control code, the design and implementation of the communications with the actuators and how MARTe integrates with the FireSignal software.

  16. Issues Involved in Developing Ada Real-Time Systems

    Science.gov (United States)

    1989-02-15

    expensive modifications to the compiler or Ada runtime system to fit a particular application. Whether we can solve the problems of programming real - time systems in...lock in solutions to problems that are not yet well understood in standards as rigorous as the Ada language. Moreover, real - time systems typically have

  17. The Synthesis of Intelligent Real-Time Systems

    Science.gov (United States)

    1990-11-09

    Synthesis of Intelligent Real - Time Systems . The purpose of the effort was to develop and extend theories and techniques that facilitate the design and...implementation of intelligent real - time systems . In particular, Teleos has extended situated-automata theory to apply to situations in which the system has

  18. Real time control of plasmas and ECRH systems on TCV

    NARCIS (Netherlands)

    Paley, J.I.; Felici, F.; Berrino, J.; Coda, S.; Cruz, N.; Duval, B.P.; Goodman, T.P.; Martin, Y.; Moret, J.-M.; Piras, F.; Rodrigues, A.P.; Santos, B.; Varandas, C.A.F.

    2008-01-01

    Developments in the real time control hardware on TCV paired with the flexibility of plasma shaping and ECRH actuators are opening many opportunities to perform real time experiments and develop algorithms and methods for fusion applications. The ability to control MHD instabilities is particularly

  19. Real time control of plasmas and ECRH systems on TCV

    NARCIS (Netherlands)

    Paley, J.I.; Berrino, J.; Coda, S.; Cruz, N.; Duval, B.P.; Felici, F.; Goodman, T.P.; Martin, Y.; Moret, J.-M.; Piras, F.; Rodriques, A.P.; Santos, B.; Varandas, C.A.F.

    2009-01-01

    Developments in the real time control hardware on Tokamak Configuration Variable (TCV) coupled with the flexibility of plasma shaping and electron cyclotron (EC) heating and current drive actuators are opening many opportunities to perform real time experiments and develop algorithms and methods for

  20. Real Time Synchronization for Creativity in Distributed Innovation Teams

    DEFF Research Database (Denmark)

    Peitersen, Dennis Kjaersgaard; Dolog, Peter; Pedersen, Esben Staunsbjerg

    2009-01-01

    In this paper we introduce a synchronization approach for real time collaborative sketching for creativity in distributed innovation teams. We base our approach on reverse AJAX. This way we ensure scalable solution for real time drawing and sketching important in creativity settings....

  1. Distributed, Embedded and Real-time Java Systems

    CERN Document Server

    Wellings, Andy

    2012-01-01

    Research on real-time Java technology has been prolific over the past decade, leading to a large number of corresponding hardware and software solutions, and frameworks for distributed and embedded real-time Java systems.  This book is aimed primarily at researchers in real-time embedded systems, particularly those who wish to understand the current state of the art in using Java in this domain.  Much of the work in real-time distributed, embedded and real-time Java has focused on the Real-time Specification for Java (RTSJ) as the underlying base technology, and consequently many of the Chapters in this book address issues with, or solve problems using, this framework. Describes innovative techniques in: scheduling, memory management, quality of service and communication systems supporting real-time Java applications; Includes coverage of multiprocessor embedded systems and parallel programming; Discusses state-of-the-art resource management for embedded systems, including Java’s real-time garbage collect...

  2. Real time ray tracing of skeletal implicit surfaces

    DEFF Research Database (Denmark)

    Rouiller, Olivier; Bærentzen, Jakob Andreas

    Modeling and rendering in real time is usually done via rasterization of polygonal meshes. We present a method to model with skeletal implicit surfaces and an algorithm to ray trace these surfaces in real time in the GPU. Our skeletal representation of the surfaces allows to create smooth models...

  3. Quo vadis? : persuasive computing using real time queue information

    NARCIS (Netherlands)

    Meys, Wouter; Groen, Maarten

    2014-01-01

    By presenting tourists with real-time information an increase in efficiency and satisfaction of their day planning can be achieved. At the same time, real-time information services can offer the municipality the opportunity to spread the tourists throughout the city centre. An important factor for

  4. Real-time communication for distributed plasma control systems

    Energy Technology Data Exchange (ETDEWEB)

    Luchetta, A. [Consorzio RFX, Associazione Euratom-ENEA sulla Fusione, Corso Stati Uniti 4, Padova 35127 (Italy)], E-mail: adriano.luchetta@igi.cnr.it; Barbalace, A.; Manduchi, G.; Soppelsa, A.; Taliercio, C. [Consorzio RFX, Associazione Euratom-ENEA sulla Fusione, Corso Stati Uniti 4, Padova 35127 (Italy)

    2008-04-15

    Real-time control applications will benefit in the near future from the enhanced performance provided by multi-core processor architectures. Nevertheless real-time communication will continue to be critical in distributed plasma control systems where the plant under control typically is distributed over a wide area. At RFX-mod real-time communication is crucial for hard real-time plasma control, due to the distributed architecture of the system, which consists of several VMEbus stations. The system runs under VxWorks and uses Gigabit Ethernet for sub-millisecond real-time communication. To optimize communication in the system, a set of detailed measurements has been carried out on the target platforms (Motorola MVME5100 and MVME5500) using either the VxWorks User Datagram Protocol (UDP) stack or raw communication based on the data link layer. Measurements have been carried out also under Linux, using its UDP stack or, in alternative, RTnet, an open source hard real-time network protocol stack. RTnet runs under Xenomai or RTAI, two popular real-time extensions based on the Linux kernel. The paper reports on the measurements carried out and compares the results, showing that the performance obtained by using open source code is suitable for sub-millisecond real-time communication in plasma control.

  5. Innovative tools for real-time simulation of dynamic systems

    NARCIS (Netherlands)

    Palli, Gianluca; Carloni, Raffaella; Melchiorri, Claudio

    2008-01-01

    In this paper, we present a software architecture, based on RTAI-Linux, for the real-time simulation of dynamic systems and for the rapid prototyping of digital controllers. Our aim is to simplify the testing phase of digital controllers by providing the real-time simulation of the plant with the

  6. Process algebra with timing: Real time and discrete time

    NARCIS (Netherlands)

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

    1999-01-01

    We present real time and discrete time versions of ACP with absolute timing and relative timing. The startingpoint is a new real time version with absolute timing, called ACPsat , featuring urgent actions and a delay operator. The discrete time versions are conservative extensions of the discrete

  7. Evaluation of the Cow Rumen Metagenome: Assembly by Single Copy Gene Analysis and Single Cell Genome Assemblies (Metagenomics Informatics Challenges Workshop: 10K Genomes at a Time)

    Energy Technology Data Exchange (ETDEWEB)

    Sczyrba, Alex

    2011-10-13

    DOE JGI's Alex Sczyrba on "Evaluation of the Cow Rumen Metagenome" and "Assembly by Single Copy Gene Analysis and Single Cell Genome Assemblies" at the Metagenomics Informatics Challenges Workshop held at the DOE JGI on October 12-13, 2011.

  8. Academic Training: Real Time Process Control - Lecture series

    CERN Multimedia

    Françoise Benz

    2004-01-01

    ACADEMIC TRAINING LECTURE REGULAR PROGRAMME 7, 8 and 9 June From 11:00 hrs to 12:00 hrs - Main Auditorium bldg. 500 Real Time Process Control T. Riesco / CERN-TS What exactly is meant by Real-time? There are several definitions of real-time, most of them contradictory. Unfortunately the topic is controversial, and there does not seem to be 100% agreement over the terminology. Real-time applications are becoming increasingly important in our daily lives and can be found in diverse environments such as the automatic braking system on an automobile, a lottery ticket system, or robotic environmental samplers on a space station. These lectures will introduce concepts and theory like basic concepts timing constraints, task scheduling, periodic server mechanisms, hard and soft real-time.ENSEIGNEMENT ACADEMIQUE ACADEMIC TRAINING Françoise Benz 73127 academic.training@cern.ch

  9. A real-time photogrammetry system based on embedded architecture

    Directory of Open Access Journals (Sweden)

    S. Y. Zheng

    2014-06-01

    Full Text Available In order to meet the demand of real-time spatial data processing and improve the online processing capability of photogrammetric system, a kind of real-time photogrammetry method is proposed in this paper. According to the proposed method, system based on embedded architecture is then designed: using FPGA, ARM+DSP and other embedded computing technology to build specialized hardware operating environment, transplanting and optimizing the existing photogrammetric algorithm to the embedded system, and finally real-time photogrammetric data processing is realized. At last, aerial photogrammetric experiment shows that the method can achieve high-speed and stable on-line processing of photogrammetric data. And the experiment also verifies the feasibility of the proposed real-time photogrammetric system based on embedded architecture. It is the first time to realize real-time aerial photogrammetric system, which can improve the online processing efficiency of photogrammetry to a higher level and broaden the application field of photogrammetry.

  10. Recent achievements in real-time computational seismology in Taiwan

    Science.gov (United States)

    Lee, S.; Liang, W.; Huang, B.

    2012-12-01

    Real-time computational seismology is currently possible to be achieved which needs highly connection between seismic database and high performance computing. We have developed a real-time moment tensor monitoring system (RMT) by using continuous BATS records and moment tensor inversion (CMT) technique. The real-time online earthquake simulation service is also ready to open for researchers and public earthquake science education (ROS). Combine RMT with ROS, the earthquake report based on computational seismology can provide within 5 minutes after an earthquake occurred (RMT obtains point source information ROS completes a 3D simulation real-time now. For more information, welcome to visit real-time computational seismology earthquake report webpage (RCS).

  11. The FERMI-Elettra distributed real-time framework

    International Nuclear Information System (INIS)

    Pivetta, L.; Gaio, G.; Passuello, R.; Scalamera, G.

    2012-01-01

    FERMI-Elettra is a Free Electron Laser (FEL) based on a 1.5 GeV linac. The pulsed operation of the accelerator and the necessity to characterize and control each electron bunch requires synchronous acquisition of the beam diagnostics together with the ability to drive actuators in real-time at the linac repetition rate. The Adeos/Xenomai real-time extensions have been adopted in order to add real-time capabilities to the Linux based control system computers running the Tango software. A software communication protocol based on Gigabit Ethernet and known as Network Reflective Memory (NRM) has been developed to implement a shared memory across the whole control system, allowing computers to communicate in real-time. The NRM architecture, the real-time performance and the integration in the control system are described. (authors)

  12. Survey of real-time processing systems for big data

    DEFF Research Database (Denmark)

    Liu, Xiufeng; Lftikhar, Nadeem; Xie, Xike

    2014-01-01

    In recent years, real-time processing and analytics systems for big data–in the context of Business Intelligence (BI)–have received a growing attention. The traditional BI platforms that perform regular updates on daily, weekly or monthly basis are no longer adequate to satisfy the fast......-changing business environments. However, due to the nature of big data, it has become a challenge to achieve the real-time capability using the traditional technologies. The recent distributed computing technology, MapReduce, provides off-the-shelf high scalability that can significantly shorten the processing time...... for big data; Its open-source implementation such as Hadoop has become the de-facto standard for processing big data, however, Hadoop has the limitation of supporting real-time updates. The improvements in Hadoop for the real-time capability, and the other alternative real-time frameworks have been...

  13. Performance evaluation of near-real-time accounting systems

    International Nuclear Information System (INIS)

    Anon.

    1981-01-01

    Examples are given illustrating the application of near-real-time accounting concepts and principles to actual nuclear facilities. Experience with prototypical systems at the AGNS reprocessing plant and the Los Alamos plutonium facility is described using examples of actual data to illustrate the performance and effectiveness of near-real-time systems. The purpose of the session is to enable participants to: (1) identify the major components of near-real-time accounting systems; (2) describe qualitatively the advantages, limitations, and performance of such systems in real nuclear facilities; (3) identify process and facility design characteristics that affect the performance of near-real-time systems; and (4) describe qualitatively the steps necessary to implement a near-real-time accounting and control system in a nuclear facility

  14. Self-Organization in Embedded Real-Time Systems

    CERN Document Server

    Brinkschulte, Uwe; Rettberg, Achim

    2013-01-01

    This book describes the emerging field of self-organizing, multicore, distributed and real-time embedded systems.  Self-organization of both hardware and software can be a key technique to handle the growing complexity of modern computing systems. Distributed systems running hundreds of tasks on dozens of processors, each equipped with multiple cores, requires self-organization principles to ensure efficient and reliable operation. This book addresses various, so-called Self-X features such as self-configuration, self-optimization, self-adaptation, self-healing and self-protection. Presents open components for embedded real-time adaptive and self-organizing applications; Describes innovative techniques in: scheduling, memory management, quality of service, communications supporting organic real-time applications; Covers multi-/many-core embedded systems supporting real-time adaptive systems and power-aware, adaptive hardware and software systems; Includes case studies of open embedded real-time self-organizi...

  15. Real-Time MENTAT programming language and architecture

    Science.gov (United States)

    Grimshaw, Andrew S.; Silberman, Ami; Liu, Jane W. S.

    1989-01-01

    Real-time MENTAT, a programming environment designed to simplify the task of programming real-time applications in distributed and parallel environments, is described. It is based on the same data-driven computation model and object-oriented programming paradigm as MENTAT. It provides an easy-to-use mechanism to exploit parallelism, language constructs for the expression and enforcement of timing constraints, and run-time support for scheduling and exciting real-time programs. The real-time MENTAT programming language is an extended C++. The extensions are added to facilitate automatic detection of data flow and generation of data flow graphs, to express the timing constraints of individual granules of computation, and to provide scheduling directives for the runtime system. A high-level view of the real-time MENTAT system architecture and programming language constructs is provided.

  16. Simultaneous detection and differentiation of dengue virus serotypes 1-4, Japanese encephalitis virus, and West Nile virus by a combined reverse-transcription loop-mediated isothermal amplification assay

    Directory of Open Access Journals (Sweden)

    Yin Jianhua

    2011-07-01

    Full Text Available Abstract Background Rapid identification and differentiation of mosquito-transmitted flaviviruses in acute-phase sera of patients and field-caught vector mosquitoes are important for the prediction and prevention of large-scale epidemics. Results We developed a flexible reverse-transcription loop-mediated isothermal amplification (RT-LAMP unit for the detection and differentiation of dengue virus serotypes 1-4 (DENV1-4, Japanese encephalitis virus (JEV, and West Nile virus (WNV. The unit efficiently amplified the viral genomes specifically at wide ranges of viral template concentrations, and exhibited similar amplification curves as monitored by a real-time PCR engine. The detection limits of the RT-LAMP unit were 100-fold higher than that of RT-PCR in 5 of the six flaviviruses. The results on specificity indicated that the six viruses in the assay had no cross-reactions with each other. By examining 66 viral strains of DENV1-4 and JEV, the unit identified the viruses with 100% accuracy and did not cross-react with influenza viruses and hantaviruses. By screening a panel of specimens containing sera of 168 patients and 279 pools of field-caught blood sucked mosquitoes, results showed that this unit is high feasible in clinical settings and epidemiologic field, and it obtained results 100% correlated with real-time RT-PCR. Conclusions The RT-LAMP unit developed in this study is able to quickly detect and accurately differentiate the six kinds of flaviviruses, which makes it extremely feasible for screening these viruses in acute-phase sera of the patients and in vector mosquitoes without the need of high-precision instruments.

  17. An In-Home Digital Network Architecture for Real-Time and Non-Real-Time Communication

    NARCIS (Netherlands)

    Scholten, Johan; Jansen, P.G.; Hanssen, F.T.Y.; Hattink, Tjalling

    2002-01-01

    This paper describes an in-home digital network architecture that supports both real-time and non-real-time communication. The architecture deploys a distributed token mechanism to schedule communication streams and to offer guaranteed quality-ofservice. Essentially, the token mechanism prevents

  18. The Fast Tracker Real Time Processor: high quality real-time tracking at ATLAS

    CERN Document Server

    Stabile, A; The ATLAS collaboration

    2011-01-01

    As the LHC luminosity is ramped up to the design level of 1x1034 cm−2 s−1 and beyond, the high rates, multiplicities, and energies of particles seen by the detectors will pose a unique challenge. Only a tiny fraction of the produced collisions can be stored on tape and immense real-time data reduction is needed. An effective trigger system must maintain high trigger efficiencies for the most important physics and at the same time suppress the enormous QCD backgrounds. This requires massive computing power to minimize the online execution time of complex algorithms. A multi-level trigger is an effective solution for an otherwise impossible problem. The Fast Tracker (FTK)[1], [2] is a proposed upgrade to the current ATLAS trigger system that will operate at full Level-1 output rates and provide high quality tracks reconstructed over the entire detector by the start of processing in Level-2. FTK is a dedicated Super Computer based on a mixture of advanced technologies. The architecture broadly employs powerf...

  19. Human immunodeficiency virus uses tRNA(Lys,3) as primer for reverse transcription in HeLa-CD4+ cells

    NARCIS (Netherlands)

    Das, A. T.; Koken, S. E.; Essink, B. B.; van Wamel, J. L.; Berkhout, B.

    1994-01-01

    Significant amounts of different tRNA molecules are present in retroviral particles, but one specific tRNA species functions as primer in reverse transcription. It is generally believed that the HIV-1 virus uses the tRNA(Lys,3) molecule as primer. This is based on sequence complementarity between

  20. Development of a rapid diagnostic assay for the detection of tomato chlorotic dwarf viroid based on isothermal reverse-transcription-recombinase polymerase amplification

    Science.gov (United States)

    A molecular diagnostic assay utilizing reverse transcription-recombinase polymerase amplification (RT-RPA) at an isothermal constant temperature of 39 °C and target-specific primers and probe were developed for the rapid, sensitive, and specific detection of tomato chlorotic dwarf viroid (TCDVd) in ...

  1. Multiplex reverse transcription-polymerase chain reaction combined with on-chip electrophoresis as a rapid screening tool for candidate gene sets

    DEFF Research Database (Denmark)

    Wittig, Rainer; Salowsky, Rüdiger; Blaich, Stephanie

    2005-01-01

    Combining multiplex reverse transcription-polymerase chain reaction (mRT-PCR) with microfluidic amplicon analysis, we developed an assay for the rapid and reliable semiquantitative expression screening of 11 candidate genes for drug resistance in human malignant melanoma. The functionality of thi...

  2. Visual Detection of Potato leafroll virus by One-step Reverse Transcription Loop-Mediated Isothermal Amplification of DNA with Hydroxynaphthol Blue Dye

    NARCIS (Netherlands)

    Ahmadi, S.; Almasi, A.M.; Fatehi, F.; Struik, P.C.; Moradi, A.

    2013-01-01

    Loop-mediated isothermal amplification (LAMP) assay is a novel technique for amplifying DNA under constant temperature, with high specificity, sensitivity, rapidity and efficiency. We applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) to visually detect Potato leafroll

  3. Multiplex Reverse Transcription-Polymerase Chain Reaction untuk Deteksi Cepat Virus Flu Burung H5N1 (MULTIPLEX REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION FOR RAPID DETECTION OF H5N1 AVIAN INFLUENZA VIRUS

    Directory of Open Access Journals (Sweden)

    Raden Wasito

    2015-05-01

    Full Text Available Avian influenza virus subtype H5N1 (AIV H5N1 is highly pathogenic and fatal in poultry. The virusis still endemic with low virulence rate, although it may play a critical role in causing high morbidity andmortality rates in poultry in Indonesia. In general, diagnostic approach for AIV H5N1 is based onconventional serological and viral isolation methods that have the potential to produce consumings oftime and relatively expensive cost within the laboratory without compromising test utility. Thus, amolecular approach of multiplex reverse transcription-polymerase chain reaction (mRT-PCR was developedand applied for the detection of matrix gene type A influenza viruses, AIV subtype subtype H5hemagglutinin gene with simultaneous detection of N1 nucleoprotein gene. Thirty sera specimens fromthe diseased commercial chickens that were specifically amplified positive-RT-PCR for AIV H5N1 wereselected for mRT-PCR. The mRT-PCR products were visualized by agarose gel electrophoresis and consistedof DNA fragments of AIV of 245 bp, 545 bp and 343 bp for M, H5 and N1 genes, respectively. Thus, themRT-PCR that can rapidly differentiate simultaneously between these genes is very important for thecontrol and even eradication of AIV transmission in poultry in Indonesia.

  4. A Process For Performance Evaluation Of Real-Time Systems

    Directory of Open Access Journals (Sweden)

    Andrew J. Kornecki

    2003-12-01

    Full Text Available Real-time developers and engineers must not only meet the system functional requirements, but also the stringent timing requirements. One of the critical decisions leading to meeting these timing requirements is the selection of an operating system under which the software will be developed and run. Although there is ample documentation on real-time systems performance and evaluation, little can be found that combines such information into an efficient process for use by developers. As the software industry moves towards clearly defined processes, creation of appropriate guidelines describing a process for performance evaluation of real-time system would greatly benefit real-time developers. This technology transition research focuses on developing such a process. PROPERT (PROcess for Performance Evaluation of Real Time systems - the process described in this paper - is based upon established techniques for evaluating real-time systems. It organizes already existing real-time performance criteria and assessment techniques in a manner consistent with a well-formed process, based on the Personal Software Process concepts.

  5. Reviewing real-time performance of nuclear reactor safety systems

    Energy Technology Data Exchange (ETDEWEB)

    Preckshot, G.G. [Lawrence Livermore National Lab., CA (United States)

    1993-08-01

    The purpose of this paper is to recommend regulatory guidance for reviewers examining real-time performance of computer-based safety systems used in nuclear power plants. Three areas of guidance are covered in this report. The first area covers how to determine if, when, and what prototypes should be required of developers to make a convincing demonstration that specific problems have been solved or that performance goals have been met. The second area has recommendations for timing analyses that will prove that the real-time system will meet its safety-imposed deadlines. The third area has description of means for assessing expected or actual real-time performance before, during, and after development is completed. To ensure that the delivered real-time software product meets performance goals, the paper recommends certain types of code-execution and communications scheduling. Technical background is provided in the appendix on methods of timing analysis, scheduling real-time computations, prototyping, real-time software development approaches, modeling and measurement, and real-time operating systems.

  6. Reviewing real-time performance of nuclear reactor safety systems

    International Nuclear Information System (INIS)

    Preckshot, G.G.

    1993-08-01

    The purpose of this paper is to recommend regulatory guidance for reviewers examining real-time performance of computer-based safety systems used in nuclear power plants. Three areas of guidance are covered in this report. The first area covers how to determine if, when, and what prototypes should be required of developers to make a convincing demonstration that specific problems have been solved or that performance goals have been met. The second area has recommendations for timing analyses that will prove that the real-time system will meet its safety-imposed deadlines. The third area has description of means for assessing expected or actual real-time performance before, during, and after development is completed. To ensure that the delivered real-time software product meets performance goals, the paper recommends certain types of code-execution and communications scheduling. Technical background is provided in the appendix on methods of timing analysis, scheduling real-time computations, prototyping, real-time software development approaches, modeling and measurement, and real-time operating systems

  7. Real-time object-oriented programming: studies and proposals

    International Nuclear Information System (INIS)

    Fouquier, Gilles

    1996-01-01

    This thesis contributes to the introduction of real-time features in object-oriented programming. Object-oriented programming favours modularity and reusability. Therefore, its application to real-time introduces many theoretical and conceptual problems. To deal with these problems, a new real-time object-oriented programming model is presented. This model is based on the active object model which allows concurrence and maintains the encapsulation property. The real-time aspect is treated by replacing the concept of task by the concept of method processing and by associating a real-time constraint to each message (priority or deadline). The set of all the running methods is scheduled. This model, called ATOME, contains several sub-models to deal with the usual concurrence control integrating their priority and deadline processing. The classical HPF and EDF scheduling avoid priority or deadline inversion. This model and its variants are new proposals to program real-time applications in the object-oriented way, therefore easing reusability and code writing. The feasibility of this approach is demonstrated by extending and existing active object-based language to real-time, in using the rules defined in the ATOME model. (author) [fr

  8. Internal control for real-time polymerase chain reaction based on MS2 bacteriophage for RNA viruses diagnostics

    Directory of Open Access Journals (Sweden)

    Miriam Ribas Zambenedetti

    Full Text Available BACKGROUND Real-time reverse transcription polymerase chain reaction (RT-PCR is routinely used to detect viral infections. In Brazil, it is mandatory the use of nucleic acid tests to detect hepatitis C virus (HCV, hepatitis B virus and human immunodeficiency virus in blood banks because of the immunological window. The use of an internal control (IC is necessary to differentiate the true negative results from those consequent from a failure in some step of the nucleic acid test. OBJECTIVES The aim of this study was the construction of virus-modified particles, based on MS2 bacteriophage, to be used as IC for the diagnosis of RNA viruses. METHODS The MS2 genome was cloned into the pET47b(+ plasmid, generating pET47b(+-MS2. MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. These particles were used as non-competitive IC in assays for RNA virus diagnostics. In addition, a competitive control for HCV diagnosis was developed by cloning a mutated HCV sequence into the MS2 replicase gene of pET47b(+-MS2, which produces a non-propagating MS2 particle. The utility of MS2-like particles as IC was evaluated in a one-step format multiplex real-time RT-PCR for HCV detection. FINDINGS We demonstrated that both competitive and non-competitive IC could be successfully used to monitor the HCV amplification performance, including the extraction, reverse transcription, amplification and detection steps, without compromising the detection of samples with low target concentrations. In conclusion, MS2-like particles generated by this strategy proved to be useful IC for RNA virus diagnosis, with advantage that they are produced by a low cost protocol. An attractive feature of this system is that it allows the construction of a multicontrol by the insertion of sequences from more than one pathogen, increasing its applicability for diagnosing different RNA viruses.

  9. Real-time embedded systems design principles and engineering practices

    CERN Document Server

    Fan, Xiaocong

    2015-01-01

    This book integrates new ideas and topics from real time systems, embedded systems, and software engineering to give a complete picture of the whole process of developing software for real-time embedded applications. You will not only gain a thorough understanding of concepts related to microprocessors, interrupts, and system boot process, appreciating the importance of real-time modeling and scheduling, but you will also learn software engineering practices such as model documentation, model analysis, design patterns, and standard conformance. This book is split into four parts to help you

  10. Real-time Testing of Operating Systems on Raspberry Pi

    OpenAIRE

    Murstad, Amund

    2016-01-01

    A subset of operating systems are called ``real time'' operating systems. While a general operating system like Windows or OSX can make no guarantee when a process gets to run, a real time one has the ability to do so. They are used for systems where the quality of the results from the operating system is not only defined by their correctness, but also by when they arrive. These kinds of systems are called Real-time Systems and can be divided into soft, hard and firm. If a deadline is missed ...

  11. Prototyping Real-Time Control in the SPS

    CERN Document Server

    Andersson, J; Jensen, L; Jones, R; Lamont, M; Wenninger, J; Wijnands, Thijs; CERN. Geneva. AB Department

    2003-01-01

    Real-time control of beam related parameters will be required in the LHC. In order to gain experience of the issues involved in implementing distributed real-time control over large distances, a prototype local orbit feedback system is being developed in the SPS. This will use 6 pickups, each equipped with the full LHC acquisition electronics chain and linked to a real-time communication and feedback system. This reports summarises the .rst tests performed with this system in October 2002, where the data from four pickups was successfully acquired and displayed at 10 Hz in the control room.

  12. Real-time Avatar Animation from a Single Image.

    Science.gov (United States)

    Saragih, Jason M; Lucey, Simon; Cohn, Jeffrey F

    2011-01-01

    A real time facial puppetry system is presented. Compared with existing systems, the proposed method requires no special hardware, runs in real time (23 frames-per-second), and requires only a single image of the avatar and user. The user's facial expression is captured through a real-time 3D non-rigid tracking system. Expression transfer is achieved by combining a generic expression model with synthetically generated examples that better capture person specific characteristics. Performance of the system is evaluated on avatars of real people as well as masks and cartoon characters.

  13. Distributed Issues for Ada Real-Time Systems

    Science.gov (United States)

    1990-07-23

    NUMBERS Distributed Issues for Ada Real - Time Systems MDA 903-87- C- 0056 S. AUTHOR(S) Thomas E. Griest 7. PERFORMING ORGANiZATION NAME(S) AND ADORESS(ES) 8...considerations. I Adding to the problem of distributed real - time systems is the issue of maintaining a common sense of time among all of the processors...because -omeone is waiting for the final output of a very large set of computations. However in real - time systems , consistent meeting of short-term

  14. Dense time discretization technique for verification of real time systems

    International Nuclear Information System (INIS)

    Makackas, Dalius; Miseviciene, Regina

    2016-01-01

    Verifying the real-time system there are two different models to control the time: discrete and dense time based models. This paper argues a novel verification technique, which calculates discrete time intervals from dense time in order to create all the system states that can be reached from the initial system state. The technique is designed for real-time systems specified by a piece-linear aggregate approach. Key words: real-time system, dense time, verification, model checking, piece-linear aggregate

  15. Formal methods for dependable real-time systems

    Science.gov (United States)

    Rushby, John

    1993-01-01

    The motivation for using formal methods to specify and reason about real time properties is outlined and approaches that were proposed and used are sketched. The formal verifications of clock synchronization algorithms are concluded as showing that mechanically supported reasoning about complex real time behavior is feasible. However, there was significant increase in the effectiveness of verification systems since those verifications were performed, at it is to be expected that verifications of comparable difficulty will become fairly routine. The current challenge lies in developing perspicuous and economical approaches to the formalization and specification of real time properties.

  16. A Distributed Computing Network for Real-Time Systems.

    Science.gov (United States)

    1980-11-03

    7 ) AU2 o NAVA TUNDEWATER SY$TEMS CENTER NEWPORT RI F/G 9/2 UIS RIBUT E 0 COMPUTIN G N LTWORK FOR REAL - TIME SYSTEMS .(U) UASSIFIED NOV Al 6 1...MORAIS - UT 92 dLEVEL c A Distributed Computing Network for Real - Time Systems . 11 𔃺-1 Gordon E/Morson I7 y tm- ,r - t "en t As J 2 -p .. - 7 I’ cNaval...NUMBER TD 5932 / N 4. TITLE mand SubotI. S. TYPE OF REPORT & PERIOD COVERED A DISTRIBUTED COMPUTING NETWORK FOR REAL - TIME SYSTEMS 6. PERFORMING ORG

  17. Developments in architecture for real-time data systems

    International Nuclear Information System (INIS)

    Heath, R.L.; Myers, W.R.

    1975-01-01

    Real-time data systems typically operate at two levels: a fast-response instrument-oriented level for data acquisition and control, and a slow human-oriented level for interaction and computation. Traditional minicomputer data systems support real-time applications by implementation of background/foreground software. Recent developments in computer technology including microprocessors enable the functional organization of hardware in distributed or hierarchical form to provide new system structures for real-time requirements. Examples of systems with distributed architecture will be discussed in detail

  18. Real-time beam profile imaging system for actinotherapy accelerator

    International Nuclear Information System (INIS)

    Lin Yong; Wang Jingjin; Song Zheng; Zheng Putang; Wang Jianguo

    2003-01-01

    This paper describes a real-time beam profile imaging system for actinotheraphy accelerator. With the flash X-ray imager and the technique of digital image processing, a real-time 3-dimension dosage image is created from the intensity profile of the accelerator beam in real time. This system helps to obtain all the physical characters of the beam in any section plane, such as FWHM, penumbra, peak value, symmetry and homogeneity. This system has been used to acquire a 3-dimension dosage distribution of dynamic wedge modulator and the transient process of beam dosage. The system configure and the tested beam profile images are also presented

  19. Expert systems for real-time monitoring and fault diagnosis

    Science.gov (United States)

    Edwards, S. J.; Caglayan, A. K.

    1989-01-01

    Methods for building real-time onboard expert systems were investigated, and the use of expert systems technology was demonstrated in improving the performance of current real-time onboard monitoring and fault diagnosis applications. The potential applications of the proposed research include an expert system environment allowing the integration of expert systems into conventional time-critical application solutions, a grammar for describing the discrete event behavior of monitoring and fault diagnosis systems, and their applications to new real-time hardware fault diagnosis and monitoring systems for aircraft.

  20. Real-Time Business Intelligence for the Utilities Industry

    Directory of Open Access Journals (Sweden)

    Janina POPEANGA

    2012-12-01

    Full Text Available In today’s competitive environment with rapid innovation in smart metering and smart grids, there is an increased need for real-time business intelligence (RTBI in the utilities industry. Giving the fact that this industry is an environment where decisions are time sensitive, RTBI solutions will help utilities improve customer experiences and operational efficiencies. The focus of this paper is on the importance of real-time business intelligence (RTBI in the utilities industry, outlining our vision of real-time business intelligence for this industry. Besides the analysis in this area, the article presents as a case study the Oracle Business Intelligence solution for utilities.

  1. The Implementation of a Real-Time Polyphase Filter

    OpenAIRE

    Adámek, Karel; Novotný, Jan; Armour, Wes

    2014-01-01

    In this article we study the suitability of dierent computational accelerators for the task of real-time data processing. The algorithm used for comparison is the polyphase filter, a standard tool in signal processing and a well established algorithm. We measure performance in FLOPs and execution time, which is a critical factor for real-time systems. For our real-time studies we have chosen a data rate of 6.5GB/s, which is the estimated data rate for a single channel on the SKAs Low Frequenc...

  2. Simultaneous Detection of Four Foodborne Viruses in Food Samples Using a One-Step Multiplex Reverse Transcription PCR.

    Science.gov (United States)

    Lee, Shin-Young; Kim, Mi-Ju; Kim, Hyun-Joong; Jeong, KwangCheol Casey; Kim, Hae-Yeong

    2018-02-28

    A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.

  3. Comparison of reverse transcription-quantitative polymerase chain reaction methods and platforms for single cell gene expression analysis.

    Science.gov (United States)

    Fox, Bridget C; Devonshire, Alison S; Baradez, Marc-Olivier; Marshall, Damian; Foy, Carole A

    2012-08-15

    Single cell gene expression analysis can provide insights into development and disease progression by profiling individual cellular responses as opposed to reporting the global average of a population. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the "gold standard" for the quantification of gene expression levels; however, the technical performance of kits and platforms aimed at single cell analysis has not been fully defined in terms of sensitivity and assay comparability. We compared three kits using purification columns (PicoPure) or direct lysis (CellsDirect and Cells-to-CT) combined with a one- or two-step RT-qPCR approach using dilutions of cells and RNA standards to the single cell level. Single cell-level messenger RNA (mRNA) analysis was possible using all three methods, although the precision, linearity, and effect of lysis buffer and cell background differed depending on the approach used. The impact of using a microfluidic qPCR platform versus a standard instrument was investigated for potential variability introduced by preamplification of template or scaling down of the qPCR to nanoliter volumes using laser-dissected single cell samples. The two approaches were found to be comparable. These studies show that accurate gene expression analysis is achievable at the single cell level and highlight the importance of well-validated experimental procedures for low-level mRNA analysis. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses

    Directory of Open Access Journals (Sweden)

    Ji Yeon Kwon

    2013-09-01

    Full Text Available A detection system based on a multiplex reverse transcription (RT polymerase chain reaction (PCR was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV, lily mottle virus (LMoV, lily symptomless virus (LSV. Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single-or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

  5. Development of Reverse Transcription Thermostable Helicase-Dependent DNA Amplification for the Detection of Tomato Spotted Wilt Virus.

    Science.gov (United States)

    Wu, Xinghai; Chen, Chanfa; Xiao, Xizhi; Deng, Ming Jun

    2016-11-01

    A protocol for the reverse transcription-helicase-dependent amplification (RT-HDA) of isothermal DNA was developed for the detection of tomato spotted wilt virus (TSWV). Specific primers, which were based on the highly conserved region of the N gene sequence in TSWV, were used for the amplification of virus's RNA. The LOD of RT-HDA, reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP), and reverse transcriptase-polymerase chain reaction (RT-PCR) assays were conducted using 10-fold serial dilution of RNA eluates. TSWV sensitivity in RT-HDA and RT-LAMP was 4 pg RNA compared with 40 pg RNA in RT-PCR. The specificity of RT-HDA for TSWV was high, showing no cross-reactivity with other tomato and Tospovirus viruses including cucumber mosaic virus (CMV), tomato black ring virus (TBRV), tomato mosaic virus (ToMV), or impatiens necrotic spot virus (INSV). The RT-HDA method is effective for the detection of TSWV in plant samples and is a potential tool for early and rapid detection of TSWV.

  6. Detection of Brazilian hantavirus by reverse transcription polymerase chain reaction amplification of N gene in patients with hantavirus cardiopulmonary syndrome

    Directory of Open Access Journals (Sweden)

    Marcos Lázaro Moreli

    2004-10-01

    Full Text Available We report a nested reverse transcription-polymerase chain reaction (RT-PCR assay for hantavirus using primers selected to match high homology regions of hantavirus genomes detected from the whole blood of hantavirus cardiopulmonary syndrome (HCPS patients from Brazil, also including the N gene nucleotide sequence of Araraquara virus. Hantavirus genomes were detected in eight out of nine blood samples from the HCPS patients by RT-PCR (88.9% positivity and in all 9 blood samples (100% positivity by nested-PCR. The eight amplicons obtained by RT-PCR (P1, P3-P9, including one obtained by nested-PCR (P-2 and not obtained by RT-PCR, were sequenced and showed high homology (94.8% to 99.1% with the N gene of Araraquara hantavirus. Although the serologic method ELISA is the most appropriate test for HCPS diagnosis, the use of nested RT-PCR for hantavirus in Brazil would contribute to the diagnosis of acute hantavirus disease detecting viral genomes in patient specimens as well as initial genomic characterization of circulating hantaviruses.

  7. Army Secure Operating System: Information Security for Real Time Systems

    National Research Council Canada - National Science Library

    Anderson, Eric

    1984-01-01

    The Army Secure Operating System (ASOS) project, under the management of the U.S. Army CECOM organization, will provide real time systems software necessary for fielding modern Battlefield Automation Systems...

  8. Real-time logo detection and tracking in video

    Science.gov (United States)

    George, M.; Kehtarnavaz, N.; Rahman, M.; Carlsohn, M.

    2010-05-01

    This paper presents a real-time implementation of a logo detection and tracking algorithm in video. The motivation of this work stems from applications on smart phones that require the detection of logos in real-time. For example, one application involves detecting company logos so that customers can easily get special offers in real-time. This algorithm uses a hybrid approach by initially running the Scale Invariant Feature Transform (SIFT) algorithm on the first frame in order to obtain the logo location and then by using an online calibration of color within the SIFT detected area in order to detect and track the logo in subsequent frames in a time efficient manner. The results obtained indicate that this hybrid approach allows robust logo detection and tracking to be achieved in real-time.

  9. Real-time earthquake monitoring: Early warning and rapid response

    Science.gov (United States)

    1991-01-01

    A panel was established to investigate the subject of real-time earthquake monitoring (RTEM) and suggest recommendations on the feasibility of using a real-time earthquake warning system to mitigate earthquake damage in regions of the United States. The findings of the investigation and the related recommendations are described in this report. A brief review of existing real-time seismic systems is presented with particular emphasis given to the current California seismic networks. Specific applications of a real-time monitoring system are discussed along with issues related to system deployment and technical feasibility. In addition, several non-technical considerations are addressed including cost-benefit analysis, public perceptions, safety, and liability.

  10. NOAA Ship Okeanos Explorer Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Okeanos Explorer Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System...

  11. NOAA Ship Nancy Foster Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Nancy Foster Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  12. NOAA Ship Gordon Gunter Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Gordon Gunter Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  13. NOAA Ship Oregon II Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Oregon II Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  14. NOAA Ship Fairweather Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Fairweather Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  15. NOAA Ship Rainier Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Rainier Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  16. NOAA Ship Pisces Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Pisces Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  17. Addressing Software Engineering Issues in Real-Time Software ...

    African Journals Online (AJOL)

    Addressing Software Engineering Issues in Real-Time Software ... systems, manufacturing process, process control, military, space exploration, and ... but also physical properties such as timeliness, Quality of Service and reliability.

  18. Real-time quasi-3D tomographic reconstruction

    Science.gov (United States)

    Buurlage, Jan-Willem; Kohr, Holger; Palenstijn, Willem Jan; Joost Batenburg, K.

    2018-06-01

    Developments in acquisition technology and a growing need for time-resolved experiments pose great computational challenges in tomography. In addition, access to reconstructions in real time is a highly demanded feature but has so far been out of reach. We show that by exploiting the mathematical properties of filtered backprojection-type methods, having access to real-time reconstructions of arbitrarily oriented slices becomes feasible. Furthermore, we present , software for visualization and on-demand reconstruction of slices. A user of can interactively shift and rotate slices in a GUI, while the software updates the slice in real time. For certain use cases, the possibility to study arbitrarily oriented slices in real time directly from the measured data provides sufficient visual and quantitative insight. Two such applications are discussed in this article.

  19. Hard-real-time resource management for autonomous spacecraft

    Science.gov (United States)

    Gat, E.

    2000-01-01

    This paper describes tickets, a computational mechanism for hard-real-time autonomous resource management. Autonomous spacecraftcontrol can be considered abstractly as a computational process whose outputs are spacecraft commands.

  20. Novel Real-Time Flight Envelope Monitoring System, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — The proposed innovation is an aircraft flight envelope monitoring system that will provide real-time in-cockpit estimations of aircraft flight envelope boundaries....

  1. RadNet-Air Near Real Time Data

    Data.gov (United States)

    U.S. Environmental Protection Agency — RadNet-Air is a national network of air monitoring stations that regularly collect air samples for near real time analysis of radioactivity. The data is transmitted...

  2. Real-time water quality monitoring and providing water quality ...

    Science.gov (United States)

    EPA and the U.S. Geological Survey (USGS) have initiated the “Village Blue” research project to provide real-time water quality monitoring data to the Baltimore community and increase public awareness about local water quality in Baltimore Harbor and the Chesapeake Bay. The Village Blue demonstration project complements work that a number of state and local organizations are doing to make Baltimore Harbor “swimmable and fishable” 2 by 2020. Village Blue is designed to build upon EPA’s “Village Green” project which provides real-time air quality information to communities in six locations across the country. The presentation, “Real-time water quality monitoring and providing water quality information to the Baltimore Community”, summarizes the Village Blue real-time water quality monitoring project being developed for the Baltimore Harbor.

  3. Real-time personal dose measurement and management system

    International Nuclear Information System (INIS)

    Zhang Zhiyong; Cheng Chang; Liu Zhengshan; Yang Huating; Deng Changming; Zhang Xiu; Guo Zhanjie

    2001-01-01

    The composition and design of a real-time personal dose measurement and management system are described. Accordingly, some pertinent hardware circuits and software codes including their operation modes are presented

  4. NOAA Ship Ronald Brown Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Ronald Brown Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  5. NOAA Ship Oscar Dyson Underway Meteorological Data, Near Real Time

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA Ship Oscar Dyson Underway Meteorological Data (Near Real Time, updated daily) are from the Shipboard Automated Meteorological and Oceanographic System (SAMOS)...

  6. Games and Scenarios for Real-Time System Validation

    DEFF Research Database (Denmark)

    Li, Shuhao

    This thesis presents research on the validation of real-time embedded software systems in the context of model-based development. The thesis proposes scenario-based and game-theoretic approaches to system analysis, verification, synthesis and testing to address the challenges that arise from....... By linking our prototype translators with existing model checker Uppaal and game solver Uppaal-Tiga, we show that these methods contribute to the interaction correctness and timeliness of early system designs. The thesis also shows that testing a real-time reactive system can be viewed as playing a timed...... communicating real-time systems can be modeled and specified with LSC. By translating LSC to timed automata (TAs), we reduce scenario-based model consistency checking and property verification to CTL real-time model checking problems, and reduce scenario-based synthesis to a timed game solving problem...

  7. Analysis and Synthesis of Distributed Real-Time Embedded Systems

    DEFF Research Database (Denmark)

    Pop, Paul; Eles, Petru; Peng, Zebo

    like automotive electronics, real-time multimedia, avionics, medical equipment, and factory systems. The proposed analysis and synthesis techniques derive optimized implementations that fulfill the imposed design constraints. An important part of the implementation process is the synthesis...

  8. The real-time price elasticity of electricity

    International Nuclear Information System (INIS)

    Lijesen, Mark G.

    2007-01-01

    The real-time price elasticity of electricity contains important information on the demand response of consumers to the volatility of peak prices. Despite the importance, empirical estimates of the real-time elasticity are hardly available. This paper provides a quantification of the real-time relationship between total peak demand and spot market prices. We find a low value for the real-time price elasticity, which may partly be explained from the fact that not all users observe the spot market price. If we correct for this phenomenon, we find the elasticity to be fairly low for consumers currently active in the spot market. If this conclusion applies to all users, this would imply a limited scope for government intervention in supply security issues. (Author)

  9. Real-Time Application Performance Steering and Adaptive Control

    National Research Council Canada - National Science Library

    Reed, Daniel

    2002-01-01

    .... The objective of the Real-time Application Performance Steering and Adaptive Control project is to replace ad hoc, post-mortem performance optimization with an extensible, portable, and distributed...

  10. Specification and Compilation of Real-Time Stream Processing Applications

    NARCIS (Netherlands)

    Geuns, S.J.

    2015-01-01

    This thesis is concerned with the specification, compilation and corresponding temporal analysis of real-time stream processing applications that are executed on embedded multiprocessor systems. An example of such applications are software defined radio applications. These applications typically

  11. Algorithm Development for a Real-Time Military Noise Monitor

    National Research Council Canada - National Science Library

    Vipperman, Jeffrey S; Bucci, Brian

    2006-01-01

    The long-range goal of this 1-year SERDP Exploratory Development (SEED) project was to create an improved real-time, high-energy military impulse noise monitoring system that can detect events with peak levels (Lpk...

  12. Real-Time Multi-Directional Equipment Site

    Data.gov (United States)

    Federal Laboratory Consortium — As part of the George E. Brown, Jr. Network for Earthquake Engineering Simulation (NEES) Program, Lehigh University has established the Real-Time Multi-Directional...

  13. Design principles of a resource sharing real-time-system

    International Nuclear Information System (INIS)

    Gliss, B.

    1978-01-01

    Criteria for developing a resource sharing real time system are given. Resource sharing necessitates extra precautions for guaranteeing stable operating conditions. Some relevant measures to insure reliability and maintainability of the system are discussed. (Auth.)

  14. Cluster Computing for Embedded/Real-Time Systems

    Science.gov (United States)

    Katz, D.; Kepner, J.

    1999-01-01

    Embedded and real-time systems, like other computing systems, seek to maximize computing power for a given price, and thus can significantly benefit from the advancing capabilities of cluster computing.

  15. A reliable information management for real-time systems

    International Nuclear Information System (INIS)

    Nishihara, Takuo; Tomita, Seiji

    1995-01-01

    In this paper, we propose a system configuration suitable for the hard realtime systems in which integrity and durability of information are important. On most hard real-time systems, where response time constraints are critical, the data which program access are volatile, and may be lost in case the systems are down. But for some real-time systems, the value-added intelligent network (IN) systems, e.g., integrity and durability of the stored data are very important. We propose a distributed system configuration for such hard real-time systems, comprised of service control modules and data management modules. The service control modules process transactions and responses based on deadline control, and the data management modules deal the stored data based on information recovery schemes well-restablished in fault real-time systems. (author)

  16. Real-time UNIX in HEP data acquisition

    International Nuclear Information System (INIS)

    Buono, S.; Gaponenko, I.; Jones, R.; Mapelli, L.; Mornacchi, G.; Prigent, D.; Sanchez-Corral, E.; Skiadelli, M.; Toppers, A.; Duval, P.Y.; Ferrato, D.; Le Van Suu, A.; Qian, Z.; Rondot, C.; Ambrosini, G.; Fumagalli, G.; Aguer, M.; Huet, M.

    1994-01-01

    Today's experimentation in high energy physics is characterized by an increasing need for sensitivity to rare phenomena and complex physics signatures, which require the use of huge and sophisticated detectors and consequently a high performance readout and data acquisition. Multi-level triggering, hierarchical data collection and an always increasing amount of processing power, distributed throughout the data acquisition layers, will impose a number of features on the software environment, especially the need for a high level of standardization. Real-time UNIX seems, today, the best solution for the platform independence, operating system interface standards and real-time features necessary for data acquisition in HEP experiments. We present the results of the evaluation, in a realistic application environment, of a Real-Time UNIX operating system: the EP/LX real-time UNIX system. ((orig.))

  17. True Time API Link (real time arrival info)

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — This link will take you to the site where you can create an account to access Port Authority's real time arrival information. To request access to Port Authority's...

  18. RadNet Real-Time Monitoring Spectrometry Data Inventory

    Data.gov (United States)

    U.S. Environmental Protection Agency — The RadNet Real-Time Monitoring Spectrometry Data Inventory contains measured data used to identify and measure specific radioactive materials in the atmosphere at...

  19. Real-Time Trajectory Generation for Autonomous Nonlinear Flight Systems

    National Research Council Canada - National Science Library

    Larsen, Michael; Beard, Randal W; McLain, Timothy W

    2006-01-01

    ... to mobile threats such as radar, jammers, and unfriendly aircraft. In Phase 1 of this STTR project, real-time path planning and trajectory generation techniques for two dimensional flight were developed and demonstrated in software simulation...

  20. Prototype real-time baseband signal combiner. [deep space network

    Science.gov (United States)

    Howard, L. D.

    1980-01-01

    The design and performance of a prototype real-time baseband signal combiner, used to enhance the received Voyager 2 spacecraft signals during the Jupiter flyby, is described. Hardware delay paths, operating programs, and firmware are discussed.

  1. Optical Real-Time Space Radiation Monitor, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Real-time dosimetry is needed to provide immediate feedback, so astronauts can minimize their exposure to ionizing radiation during periods of high solar activity....

  2. Design a Fault Tolerance for Real Time Distributed System

    OpenAIRE

    Ban M. Khammas

    2012-01-01

    This paper designed a fault tolerance for soft real time distributed system (FTRTDS). This system is designed to be independently on specific mechanisms and facilities of the underlying real time distributed system. It is designed to be distributed on all the computers in the distributed system and controlled by a central unit.Besides gathering information about a target program spontaneously, it provides information about the target operating system and the target hardware in order to diagno...

  3. Wireless Sensor Network Metrics for Real-Time Systems

    Science.gov (United States)

    2009-05-20

    Wireless Sensor Network Metrics for Real-Time Systems Phoebus Wei-Chih Chen Electrical Engineering and Computer Sciences University of California at...3. DATES COVERED 00-00-2009 to 00-00-2009 4. TITLE AND SUBTITLE Wireless Sensor Network Metrics for Real-Time Systems 5a. CONTRACT NUMBER 5b... wireless sensor networks (WSNs) is moving from studies of WSNs in isolation toward studies where the WSN is treated as a component of a larger system

  4. Implementation of real-time duplex synthetic aperture ultrasonography

    DEFF Research Database (Denmark)

    Hemmsen, Martin Christian; Larsen, Lee; Kjeldsen, Thomas

    2015-01-01

    This paper presents a real-time duplex synthetic aperture imaging system, implemented on a commercially available tablet. This includes real-time wireless reception of ultrasound signals and GPU processing for B-mode and Color Flow Imaging (CFM). The objective of the work is to investigate the im...... and that the required bandwidth between the probe and processing unit is within the current Wi-Fi standards....

  5. Classification and overview of research in real-time imaging

    Science.gov (United States)

    Sinha, Purnendu; Gorinsky, Sergey V.; Laplante, Phillip A.; Stoyenko, Alexander D.; Marlowe, Thomas J.

    1996-10-01

    Real-time imaging has application in areas such as multimedia, virtual reality, medical imaging, and remote sensing and control. Recently, the imaging community has witnessed a tremendous growth in research and new ideas in these areas. To lend structure to this growth, we outline a classification scheme and provide an overview of current research in real-time imaging. For convenience, we have categorized references by research area and application.

  6. Novel methods for real-time 3D facial recognition

    OpenAIRE

    Rodrigues, Marcos; Robinson, Alan

    2010-01-01

    In this paper we discuss our approach to real-time 3D face recognition. We argue the need for real time operation in a realistic scenario and highlight the required pre- and post-processing operations for effective 3D facial recognition. We focus attention to some operations including face and eye detection, and fast post-processing operations such as hole filling, mesh smoothing and noise removal. We consider strategies for hole filling such as bilinear and polynomial interpolation and Lapla...

  7. Limited Preemptive Scheduling in Real-time Systems

    OpenAIRE

    Thekkilakattil, Abhilash

    2016-01-01

    Preemptive and non-preemptive scheduling paradigms typically introduce undesirable side effects when scheduling real-time tasks, mainly in the form of preemption overheads and blocking, that potentially compromise timeliness guarantees. The high preemption overheads in preemptive real-time scheduling may imply high resource utilization, often requiring significant over-provisioning, e.g., pessimistic Worst Case Execution Time (WCET) approximations. Non-preemptive scheduling, on the other hand...

  8. Real-time measurement and control at Jet. Experiment Control

    International Nuclear Information System (INIS)

    Felton, R.; Zabeo, L.; Sartori, F.; Piccolo, F.; Farthing, J.; Budd, T.; Dorling, S.; McCullen, P.; Harling, J.; Dalley, S.; Goodyear, A.; Stephen, A.; Card, P.; Bright, M.; Lucock, R.; Jones, E.; Griph, S.; Hogben, C.; Beldishevski, M.; Buckley, M.; Davis, J.; Young, I.; Hemming, O.; Wheatley, M.; Heesterman, P.; Lloyd, G.; Walters, M.; Bridge, R.; Leggate, H.; Howell, D.; Zastrow, K.D.; Giroud, C.; Coffey, I.; Hawkes, N.; Stamp, M.; Barnsley, R.; Edlington, T.; Guenther, K.; Gowers, C.; Popovichef, S.; Huber, A.; Ingesson, C.; Joffrin, E.; Mazon, D.; Moreau, D.; Murari, A.; Riva, M.; Barana, O.; Bolzonella, T.; Valisa, M.; Innocente, P.; Zerbini, M.; Bosak, K.; Blum, J.; Vitale, E.; Crisanti, F.; La Luna, E. de; Sanchez, J.

    2004-01-01

    Over the past few ears, the preparation of ITER-relevant plasma scenarios has been the main focus experimental activity on tokamaks. The development of integrated, simultaneous, real-time controls of plasma shape, current, pressure, temperature, radiation, neutron profiles, and also impurities, ELMs (edge localized modes) and MHD are now seen to be essential for further development of quasi-steady state conditions with feedback, or the stabilisation of transient phenomena with event-driven actions. For this thrust, the EFDA JET Real Time Project has developed a set of real-time plasma measurements, experiment control, and communication facilities. The Plasma Diagnostics used for real-time experiments are Far Infra Red interferometry, polarimetry, visible, UV and X-ray spectroscopy, LIDAR, bolometry, neutron and magnetics. Further analysis systems produce integrated results such as temperature profiles on geometry derived from MHD equilibrium solutions. The Actuators include toroidal, poloidal and divertor coils, gas and pellet fuelling, neutral beam injection, radiofrequency (ICRH) waves and microwaves (LH). The Heating/Fuelling Operators can either define a power or gas request waveform or select the real-time instantaneous power/gas request from the Real Time Experiment Central Control (RTCC) system. The Real Time Experiment Control system provides both a high-level, control-programming environment and interlocks with the actuators. A MATLAB facility is being developed for the development of more complex controllers. The plasma measurement, controller and plant control systems communicate in ATM network. The EFDA Real Time project is essential groundwork for future reactors such as ITER. It involves many staff from several institutions. The facility is now frequently used in experiments. (authors)

  9. Research and application of embedded real-time operating system

    Science.gov (United States)

    Zhang, Bo

    2013-03-01

    In this paper, based on the analysis of existing embedded real-time operating system, the architecture of an operating system is designed and implemented. The experimental results show that the design fully complies with the requirements of embedded real-time operating system, can achieve the purposes of reducing the complexity of embedded software design and improving the maintainability, reliability, flexibility. Therefore, this design program has high practical value.

  10. Real-time fault diagnosis and fault-tolerant control

    OpenAIRE

    Gao, Zhiwei; Ding, Steven X.; Cecati, Carlo

    2015-01-01

    This "Special Section on Real-Time Fault Diagnosis and Fault-Tolerant Control" of the IEEE Transactions on Industrial Electronics is motivated to provide a forum for academic and industrial communities to report recent theoretic/application results in real-time monitoring, diagnosis, and fault-tolerant design, and exchange the ideas about the emerging research direction in this field. Twenty-three papers were eventually selected through a strict peer-reviewed procedure, which represent the mo...

  11. Multiprocessor Real-Time Scheduling with Hierarchical Processor Affinities

    OpenAIRE

    Bonifaci , Vincenzo; Brandenburg , Björn; D'Angelo , Gianlorenzo; Marchetti-Spaccamela , Alberto

    2016-01-01

    International audience; Many multiprocessor real-time operating systems offer the possibility to restrict the migrations of any task to a specified subset of processors by setting affinity masks. A notion of " strong arbitrary processor affinity scheduling " (strong APA scheduling) has been proposed; this notion avoids schedulability losses due to overly simple implementations of processor affinities. Due to potential overheads, strong APA has not been implemented so far in a real-time operat...

  12. Overview of real-time operating systems on microprocessor platforms

    International Nuclear Information System (INIS)

    Luong, T.T.

    1994-01-01

    This paper attempts to overview the real-time operating systems on microprocessor platforms in the field of experimental physics facility controls. The key issues regarding operating systems as well as standards and development environment are discussed. As an illustration, some current industrial products are indicated. Also, real-time systems operating in some institutes of the EPS/EPCS inter divisional group are reviewed. (author). 3 refs., 4 figs

  13. Boat, wake, and wave real-time simulation

    Science.gov (United States)

    Świerkowski, Leszek; Gouthas, Efthimios; Christie, Chad L.; Williams, Owen M.

    2009-05-01

    We describe the extension of our real-time scene generation software VIRSuite to include the dynamic simulation of small boats and their wakes within an ocean environment. Extensive use has been made of the programmabilty available in the current generation of GPUs. We have demonstrated that real-time simulation is feasible, even including such complexities as dynamical calculation of the boat motion, wake generation and calculation of an FFTgenerated sea state.

  14. Processor tradeoffs in distributed real-time systems

    Science.gov (United States)

    Krishna, C. M.; Shin, Kang G.; Bhandari, Inderpal S.

    1987-01-01

    The problem of the optimization of the design of real-time distributed systems is examined with reference to a class of computer architectures similar to the continuously reconfigurable multiprocessor flight control system structure, CM2FCS. Particular attention is given to the impact of processor replacement and the burn-in time on the probability of dynamic failure and mean cost. The solution is obtained numerically and interpreted in the context of real-time applications.

  15. Diagnosis of aerobic vaginitis by quantitative real-time PCR

    OpenAIRE

    Rumyantseva, T. A.; Bellen, G.; Savochkina, Y. A.; Guschin, A. E.; Donders, G.G.G.

    2016-01-01

    Abstract: Purpose To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. Methods Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. Resu...

  16. Real-time radiography at the NECTAR facility

    International Nuclear Information System (INIS)

    Buecherl, T.; Lierse von Gostomski, Ch.

    2011-01-01

    A feasibility study has shown that real-time radiography using fission neutrons is possible at the NECTAR facility, when using an improved detection system for fast variations (Buecherl et al., 2009 ). Continuing this study, real-time measurements of slowly varying processes like the water uptake in medium sized trunks (diameter about 12 cm) and of slow periodic processes (e.g. a slowly rotating iron disk) are investigated successfully using the existing detection system.

  17. Real-time radiography at the NECTAR facility

    Science.gov (United States)

    Bücherl, T.; Lierse von Gostomski, Ch.

    2011-09-01

    A feasibility study has shown that real-time radiography using fission neutrons is possible at the NECTAR facility, when using an improved detection system for fast variations (Bücherl et al., 2009 [1]). Continuing this study, real-time measurements of slowly varying processes like the water uptake in medium sized trunks (diameter about 12 cm) and of slow periodic processes (e.g. a slowly rotating iron disk) are investigated successfully using the existing detection system.

  18. Real-time radiography at the NECTAR facility

    Energy Technology Data Exchange (ETDEWEB)

    Buecherl, T., E-mail: thomas.buecherl@radiochemie.de [Lehrstuhl fuer Radiochemie (RCM), Technische Universitaet Muenchen (TUM), Walther-Meissner-Str. 3, 85748 Garching (Germany); Lierse von Gostomski, Ch. [Lehrstuhl fuer Radiochemie (RCM), Technische Universitaet Muenchen (TUM), Walther-Meissner-Str. 3, 85748 Garching (Germany)

    2011-09-21

    A feasibility study has shown that real-time radiography using fission neutrons is possible at the NECTAR facility, when using an improved detection system for fast variations (Buecherl et al., 2009 ). Continuing this study, real-time measurements of slowly varying processes like the water uptake in medium sized trunks (diameter about 12 cm) and of slow periodic processes (e.g. a slowly rotating iron disk) are investigated successfully using the existing detection system.

  19. Magnetoresistive sensor for real-time single nucleotide polymorphism genotyping

    DEFF Research Database (Denmark)

    Rizzi, Giovanni; Østerberg, Frederik Westergaard; Dufva, Martin

    2014-01-01

    We demonstrate a magnetoresistive sensor platform that allows for the real-time detection of point mutations in DNA targets. Specifically, we detect point mutations at two sites in the human beta globin gene. For DNA detection, the present sensor technology has a detection limit of about 160p...... of magnetic beads, which enables real-time quantification of the specific binding of magnetic beads to the sensor surface under varying experimental conditions....

  20. Real-time flight test data distribution and display

    Science.gov (United States)

    Nesel, Michael C.; Hammons, Kevin R.

    1988-01-01

    Enhancements to the real-time processing and display systems of the NASA Western Aeronautical Test Range are described. Display processing has been moved out of the telemetry and radar acquisition processing systems super-minicomputers into user/client interactive graphic workstations. Real-time data is provided to the workstations by way of Ethernet. Future enhancement plans include use of fiber optic cable to replace the Ethernet.