Real-time temperature field measurement based on acoustic tomography

International Nuclear Information System (INIS)

Bao, Yong; Jia, Jiabin; Polydorides, Nick

2017-01-01

Acoustic tomography can be used to measure the temperature field from the time-of-flight (TOF). In order to capture real-time temperature field changes and accurately yield quantitative temperature images, two improvements to the conventional acoustic tomography system are studied: simultaneous acoustic transmission and TOF collection along multiple ray paths, and an offline iteration reconstruction algorithm. During system operation, all the acoustic transceivers send modulated and filtered wideband Kasami sequences simultaneously to facilitate fast and accurate TOF measurements using cross-correlation detection. For image reconstruction, the iteration process is separated and executed offline beforehand to shorten computation time for online temperature field reconstruction. The feasibility and effectiveness of the developed methods are validated in the simulation study. The simulation results demonstrate that the proposed method can reduce the processing time per frame from 160 ms to 20 ms, while the reconstruction error remains less than 5%. Hence, the proposed method has great potential in the measurement of rapid temperature change with good temporal and spatial resolution. (paper)

Real-time deformation measurement using a transportable shearography system

Science.gov (United States)

Weijers, A. L.; van Brug, Hedser H.; Frankena, Hans J.

1997-03-01

A new system for deformation visualization has been developed, being a real time phase stepped shearing speckle interferometer. This system provides the possibility to measure quantitatively deformations of diffusely reflecting objects in an industrial environment. The main characteristics of this interferometer are its speed of operation and its reduced sensitivity to external disturbances. Apart from its semiconductor laser source, this system has a shoe-box size and is mounted on a tripod for easy handling during inspection. This paper describes the shearing speckle interferometry set-up, as it is developed at our laboratory and its potential for detecting defects.

Feasibility of iFlow color-coding technique in quantitative real-time measurement of hemodynamic changes after transarterial chemoembolization for hepatocellular carcinoma

Directory of Open Access Journals (Sweden)

WANG Yuzhe

2018-01-01

Full Text Available Objective To investigate the value of iFlow color-coding technique in quantitative real-time analysis of hemodynamic changes after transarterial chemoembolization (TACE for hepatocellular carcinoma (HCC. Methods A total of 31 patients who were diagnosed with HCC in Shanghai Fifth People′s Hospital from December 2015 to January 2017 were enrolled. No patient underwent surgical operation or ablation. All patients underwent TACE with the same contrast agent, high-pressure injector parameters, and place of angiographic catheter. The iFlow technique was used to generate two-dimensional color-coded images and time-density curve (TDC before and after surgery and measure the opening of the angiographic catheter and the time to peak (TTP of the starting and ending points of the major tumor feeding arteries, as well as the ratio of the areas under the curve (AUC of TDC of tumor tissue and the opening of the angiographic catheter. The paired t-test was used for comparison of continuous data between groups. Results TTP of the major tumor feeding arteries was 4.64±0.49 s before TACE and 5.97±0.84 s after TACE (t=11.57, P＜0.01, and there was a significant difference in AUC between the tumor tissue and the opening of the angiographic catheter (0.53±0.15 vs 0.16±0.12, t=25.85, P＜0.01. There was no significant difference in TTP between the opening of the angiographic catheter and the major tumor feeding arteries before and after TACE (P＞0.05. Before TACE, the TDC of tumor feeding arteries had a shape of “rapid increase-rapid reduction” with relatively high slope and peak value, while after TACE, the TDC had a shape of “increase-flat-reduction” with reductions in slope and peak value. Conclusion The iFlow technique can perform real-time measurement of TTP and TDC of the region of interest and helps with quantitative evaluation of hemodynamic changes in HCC. Therefore, it can provide objective quantitative indices for evaluating the degree of

Coalescence measurements for evolving foams monitored by real-time projection imaging

International Nuclear Information System (INIS)

Myagotin, A; Helfen, L; Baumbach, T

2009-01-01

Real-time radiographic projection imaging together with novel spatio-temporal image analysis is presented to be a powerful technique for the quantitative analysis of coalescence processes accompanying the generation and temporal evolution of foams and emulsions. Coalescence events can be identified as discontinuities in a spatio-temporal image representing a sequence of projection images. Detection, identification of intensity and localization of the discontinuities exploit a violation criterion of the Fourier shift theorem and are based on recursive spatio-temporal image partitioning. The proposed method is suited for automated measurements of discontinuity rates (i.e., discontinuity intensity per unit time), so that large series of radiographs can be analyzed without user intervention. The application potential is demonstrated by the quantification of coalescence during the formation and decay of metal foams monitored by real-time x-ray radiography

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

Quantitative analysis of diet structure by real-time PCR, reveals different feeding patterns by two dominant grasshopper species

Science.gov (United States)

Huang, Xunbing; Wu, Huihui; McNeill, Mark Richard; Qin, Xinghu; Ma, Jingchuan; Tu, Xiongbing; Cao, Guangchun; Wang, Guangjun; Nong, Xiangqun; Zhang, Zehua

2016-01-01

Studies on grasshopper diets have historically employed a range of methodologies, each with certain advantages and disadvantages. For example, some methodologies are qualitative instead of quantitative. Others require long experimental periods or examine population-level effects, only. In this study, we used real-time PCR to examine diets of individual grasshoppers. The method has the advantage of being both fast and quantitative. Using two grasshopper species, Oedaleus asiaticus and Dasyhippus barbipes, we designed ITS primer sequences for their three main host plants, Stipa krylovii, Leymus chinensis and Cleistogenes squarrosa and used real-time PCR method to test diet structure both qualitatively and quantitatively. The lowest detection efficiency of the three grass species was ~80% with a strong correlation between actual and PCR-measured food intake. We found that Oedaleus asiaticus maintained an unchanged diet structure across grasslands with different grass communities. By comparison, Dasyhippus barbipes changed its diet structure. These results revealed why O. asiaticus distribution is mainly confined to Stipa-dominated grassland, and D. barbipes is more widely distributed across Inner Mongolia. Overall, real-time PCR was shown to be a useful tool for investigating grasshopper diets, which in turn offers some insight into grasshopper distributions and improved pest management. PMID:27562455

Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR

NARCIS (Netherlands)

Boutaga, Khalil; van Winkelhoff, Arie Jan; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

2005-01-01

Periodontitis is a multi-factorial chronic inflammatory and destructive disease of the tooth-supporting tissues. Quantitative anaerobic culture techniques have been used for microbial diagnosis of the different forms of the disease. The aim of this study was to compare real-time PCR with

Selection of reference genes for quantitative real-time PCR in bovine preimplantation embryos

Directory of Open Access Journals (Sweden)

Van Zeveren Alex

2005-12-01

Full Text Available Abstract Background Real-time quantitative PCR is a sensitive and very efficient technique to examine gene transcription patterns in preimplantation embryos, in order to gain information about embryo development and to optimize assisted reproductive technologies. Critical to the succesful application of real-time PCR is careful assay design, reaction optimization and validation to maximize sensitivity and accuracy. In most of the studies published GAPD, ACTB or 18S rRNA have been used as a single reference gene without prior verification of their expression stability. Normalization of the data using unstable controls can result in erroneous conclusions, especially when only one reference gene is used. Results In this study the transcription levels of 8 commonly used reference genes (ACTB, GAPD, Histone H2A, TBP, HPRT1, SDHA, YWHAZ and 18S rRNA were determined at different preimplantation stages (2-cell, 8-cell, blastocyst and hatched blastocyst in order to select the most stable genes to normalize quantitative data within different preimplantation embryo stages. Conclusion Using the geNorm application YWHAZ, GAPD and SDHA were found to be the most stable genes across the examined embryonic stages, while the commonly used ACTB was shown to be highly regulated. We recommend the use of the geometric mean of those 3 reference genes as an accurate normalization factor, which allows small expression differences to be reliably measured.

Intra-laboratory validation of chronic bee paralysis virus quantitation using an accredited standardised real-time quantitative RT-PCR method.

Science.gov (United States)

Blanchard, Philippe; Regnault, Julie; Schurr, Frank; Dubois, Eric; Ribière, Magali

2012-03-01

Chronic bee paralysis virus (CBPV) is responsible for chronic bee paralysis, an infectious and contagious disease in adult honey bees (Apis mellifera L.). A real-time RT-PCR assay to quantitate the CBPV load is now available. To propose this assay as a reference method, it was characterised further in an intra-laboratory study during which the reliability and the repeatability of results and the performance of the assay were confirmed. The qPCR assay alone and the whole quantitation method (from sample RNA extraction to analysis) were both assessed following the ISO/IEC 17025 standard and the recent XP U47-600 standard issued by the French Standards Institute. The performance of the qPCR assay and of the overall CBPV quantitation method were validated over a 6 log range from 10(2) to 10(8) with a detection limit of 50 and 100 CBPV RNA copies, respectively, and the protocol of the real-time RT-qPCR assay for CBPV quantitation was approved by the French Accreditation Committee. Copyright © 2011 Elsevier B.V. All rights reserved.

Real-time elastography with a novel quantitative technology for assessment of liver fibrosis in chronic hepatitis B

International Nuclear Information System (INIS)

Wang Juan; Guo Long; Shi Xiuying; Pan Wenqian; Bai Yunfei; Ai Hong

2012-01-01

Background: The accurate evaluation of liver fibrosis stage is important in determining the treatment strategy. The limitations of percutaneous liver biopsy as the gold standard are obvious for invasion. Real-time elastography with conventional ultrasound probes and a new quantitative technology for diffuse histological lesion is a novel approach for staging of liver fibrosis. Purpose: This study aimed to evaluate the value of real-time tissue elastography with a new quantitative technology for the assessment of liver fibrosis stage. Materials and methods: Real-time elastography was performed in 55 patients with liver fibrosis and chronic hepatitis B and in 20 healthy volunteers. Eleven parameters for every patient in colorcode image obtained from the real-time elastography were analyzed with principal components analysis. We analyzed the correlation between elasticity index and liver fibrosis stage and the accuracy of real-time elastography for liver fibrosis staging. Additionally, aspartate transaminase-to-platelet ratio index was also included in the analysis. Results: The Spearman's correlation coefficient between the elasticity index and the histologic fibrosis stage was 0.81, which is highly significant (p 0.05), respectively. Conclusions: Real-time elastography with a new quantitative technology for diffuse histological lesion is a new and promising sonography-based noninvasive method for the assessment of liver fibrosis in patients with chronic hepatitis B.

The IPERMOB System for Effective Real-Time Road Travel Time Measurement and Prediction

OpenAIRE

Martelli, Francesca; Renda, Maria Elena; Santi, Paolo

2010-01-01

Accurate, real-time measurement and estimation of road travel time is considered a central problem in the design of advanced Intelligent Transportation Systems. In particular, whether eective, real-time collection of travel time measurements in a urban area is possible is, to the best of our knowledge, still an open problem. In this paper, we introduce the IPERMOB system for efficient, real-time collection of travel time measurements in urban areas through vehicular networks. We demonstrate t...

Development of real-time PCR for detection and quantitation of Streptococcus parauberis.

Science.gov (United States)

Nguyen, T L; Lim, Y J; Kim, D-H; Austin, B

2016-01-01

Streptococcus parauberis is an increasing threat to aquaculture of olive flounder, Paralichthys olivaceus Temminck & Schlegel, in South Korea. We developed a real-time polymerase chain reaction (PCR) method using the TaqMan probe assay to detect and quantify S. parauberis by targeting the gyrB gene sequences, which are effective for molecular analysis of the genus Streptococcus. Our real-time PCR assay is capable of detecting 10 fg of genomic DNA per reaction. The intra- and interassay coefficient of variation (CV) values ranged from 0.42-1.95%, demonstrating that the assay has good reproducibility. There was not any cross-reactivity to Streptococcus iniae or to other streptococcal/lactococcal fish pathogens, such as S. agalactiae and Lactococcus garvieae, indicating that the assay is highly specific to S. parauberis. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. parauberis from inoculated tissue homogenates (r = 0.957; P < 0.05). Hence, this sensitive and specific real-time PCR is a valuable tool for diagnostic quantitation of S. parauberis in clinical samples. © 2014 John Wiley & Sons Ltd.

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.

Detection of medically important Candida species by absolute quantitation real-time polymerase chain reaction.

Science.gov (United States)

Than, Leslie Thian Lung; Chong, Pei Pei; Ng, Kee Peng; Seow, Heng Fong

2015-01-01

The number of invasive candidiasis cases has risen especially with an increase in the number of immunosuppressed and immunocom promised patients. The early detection of Candida species which is specific and sensitive is important in determining the correct administration of antifungal drugs to patients. This study aims to develop a method for the detection, identification and quantitation of medically important Candida species through quantitative polymerase chain reaction (qPCR). The isocitrate lyase (ICL) gene which is not found in mammals was chosen as the target gene of real-time PCR. Absolute quantitation of the gene copy number was achieved by constructing the plasmid containing the ICL gene which is used to generate standard curve. Twenty fungal species, two bacterial species and human DNA were tested to check the specificity of the detection method. All eight Candida species were successfully detected, identified and quantitated based on the ICL gene. A seven-log range of the gene copy number and a minimum detection limit of 10(3) copies were achieved. A one-tube absolute quantification real-time PCR that differentiates medically important Candida species via individual unique melting temperature was achieved. Analytical sensitivity and specificity were not compromised.

Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant (Camellia sinensis (L.) O. Kuntze)

Science.gov (United States)

Quantitative real-time polymerase chain reaction (qRT-PCR) is a commonly used technique for measuring gene expression levels due to its simplicity, specificity, and sensitivity. Reliable reference selection for the accurate quantification of gene expression under various experimental conditions is a...

Statistical aspects of quantitative real-time PCR experiment design.

Science.gov (United States)

Kitchen, Robert R; Kubista, Mikael; Tichopad, Ales

2010-04-01

Experiments using quantitative real-time PCR to test hypotheses are limited by technical and biological variability; we seek to minimise sources of confounding variability through optimum use of biological and technical replicates. The quality of an experiment design is commonly assessed by calculating its prospective power. Such calculations rely on knowledge of the expected variances of the measurements of each group of samples and the magnitude of the treatment effect; the estimation of which is often uninformed and unreliable. Here we introduce a method that exploits a small pilot study to estimate the biological and technical variances in order to improve the design of a subsequent large experiment. We measure the variance contributions at several 'levels' of the experiment design and provide a means of using this information to predict both the total variance and the prospective power of the assay. A validation of the method is provided through a variance analysis of representative genes in several bovine tissue-types. We also discuss the effect of normalisation to a reference gene in terms of the measured variance components of the gene of interest. Finally, we describe a software implementation of these methods, powerNest, that gives the user the opportunity to input data from a pilot study and interactively modify the design of the assay. The software automatically calculates expected variances, statistical power, and optimal design of the larger experiment. powerNest enables the researcher to minimise the total confounding variance and maximise prospective power for a specified maximum cost for the large study. Copyright 2010 Elsevier Inc. All rights reserved.

Diagnosis of aerobic vaginitis by quantitative real-time PCR.

Science.gov (United States)

Rumyantseva, T A; Bellen, G; Savochkina, Y A; Guschin, A E; Donders, G G G

2016-07-01

To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. 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. Some level of AV was diagnosed in 23 (23.7 %) cases. Various concentrations of Enterobacteriacea, Staphylococcus spp., Streptococcus spp. were detected an all patients. Enterococcus spp. were detected in 76 (78.3 %) cases. Summarized concentrations of aerobes were tenfold higher in AV-positive compared to AV-negative cases [7.30lg vs 6.06lg (p = 0.02)]. Concentrations of aerobes in severe, moderate and light AV cases did not vary significantly (p = 0.14). Concentration of lactobacilli was 1000-fold lower in AV-positive cases compared to normal cases (5.3lg vs 8.3lg, p AV-positive cases [19/22 (86.4 %) samples]. The relation of high loads of aerobes to the low numbers of Lactobacilli are a reliable marker for the presence of AV and could substitute microscopy as a test. PCR may be a good standardized substitution for AV diagnosis in settings where well-trained microscopists are lacking.

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)

Embryonation of Ostertagia ostertagi eggs affects the outcome of real-time quantitative PCR

DEFF Research Database (Denmark)

Drag, Markus; Höglund, Johan; Nejsum, Peter

prior to detection and quantification by real-time quantitative polymerase chain reaction (qPCR). Fresh O. ostertagi eggs were isolated from cattle faeces and stored at 4°C or 25°C under aerobic or anaerobic conditions. Embryonation was monitored by microscopy and the ITS2 copies were determined by q...... the outcome of qPCR analysis for the quantitative determination of O. ostertagi eggs in cattle faeces. Cold storage at 4°C for up to 3 days or anaerobicvacuum packing at 25°C for up to 336 h will entail no undesirable effects on ITS2 copies....

Embryonation of Ostertagia ostertagi eggs affects the outcome of real-time quantitative PCR

DEFF Research Database (Denmark)

Drag, Markus; Höglund, Johan; Nejsum, Peter

prior to detection and quantification by real-time quantitative polymerase chain reaction (qPCR) . Fresh O. ostertagi eggs were isolated from cattle faeces and stored at 4°C or 25°C under aerobic or anaerobic conditions. Embryonation was monitored by microscopy and the ITS2 copies were determined by q...... the outcome of qPCR analysis for the quantitative determination of O. ostertagi eggs in cattle faeces. Cold storage at 4°C for up to 3 days or anaerobic vacuum packing at 25°C for up to 336 h will entail no undesirable effects on ITS2 copies....

Comparative evaluation of a laboratory developed real-time PCR assay and the RealStar® HHV-6 PCR Kit for quantitative detection of human herpesvirus 6.

Science.gov (United States)

Yip, Cyril C Y; Sridhar, Siddharth; Cheng, Andrew K W; Fung, Ami M Y; Cheng, Vincent C C; Chan, Kwok-Hung; Yuen, Kwok-Yung

2017-08-01

HHV-6 reactivation in immunocompromised patients is common and may be associated with serious morbidity and mortality; therefore, early detection and initiation of therapy might be of benefit. Real-time PCR assays allow for early identification of HHV-6 reactivation to assist in providing a timely response. Thus, we compared the performance of an in-house developed HHV-6 quantitative PCR assay with a commercially available kit, the RealStar ® HHV-6 PCR Kit. The analytical sensitivity, analytical specificity, linearity, precision and accuracy of the in-house developed HHV-6 qPCR assay were evaluated. The diagnostic performance of the in-house HHV-6 qPCR assay was compared with the RealStar ® HHV-6 PCR Kit, using 72 clinical specimens and 17 proficiency testing samples. Linear regression analysis of the quantitative results showed a dynamic range from 2 to 10 log 10 copies/ml and a coefficient of determination (R 2 ) of 0.999 for the in-house assay. A dilution series demonstrated a limit of detection and a limit of quantification of 1.7 log 10 and 2 log 10 copies/ml, respectively. The precision of the assay was highly reproducible among runs with coefficients of variance (CV) ranging from 0.27% to 4.37%. A comparison of 27 matched samples showed an excellent correlation between the quantitative viral loads measured by the in-house HHV-6 qPCR assay and the RealStar ® HHV-6 PCR Kit (R 2 =0.926; PPCR Kit. Copyright © 2017 Elsevier B.V. All rights reserved.

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.

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

Real-time hostile attribution measurement and aggression in children.

Science.gov (United States)

Yaros, Anna; Lochman, John E; Rosenbaum, Jill; Jimenez-Camargo, Luis Alberto

2014-01-01

Hostile attributions are an important predictor of aggression in children, but few studies have measured hostile attributions as they occur in real-time. The current study uses an interactive video racing game to measure hostile attributions while children played against a presumed peer. A sample of 75 children, ages 10-13, used nonverbal and verbal procedures to respond to ambiguous provocation by their opponent. Hostile attributions were significantly positively related to parent-rated reactive aggression, when controlling for proactive aggression. Hostile attributions using a nonverbal response procedure were negatively related to proactive aggression, when controlling for reactive aggression. Results suggest hostile attributions in real-time occur quickly and simultaneously with social interaction, which differs from the deliberative, controlled appraisals measured with vignette-based instruments. The relation between real-time hostile attributions and reactive aggression could be accounted for by the impulsive response style that is characteristic of reactive aggression, whereas children exhibiting proactive aggression may be more deliberate and intentional in their responding, resulting in a negative relation with real-time hostile attributions. These findings can be used both to identify children at risk for aggression and to enhance preventive interventions. © 2014 Wiley Periodicals, Inc.

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

Directory of Open Access Journals (Sweden)

Ana Lisa do Vale Gomes

2006-10-01

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

Data Acceptance Criteria for Standardized Human-Associated Fecal Source Identification Quantitative Real-Time PCR Methods

Science.gov (United States)

There is a growing interest in the application of human-associated fecal sourceidentification quantitative real-time PCR (qPCR) technologies for water quality management. The transition from a research tool to a standardized protocol requires a high degree of confidence in data q...

Mathematical analysis of the real time array PCR (RTA PCR) process

NARCIS (Netherlands)

Dijksman, Johan Frederik; Pierik, A.

2012-01-01

Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the

Evaluation of airway protection: Quantitative timing measures versus penetration/aspiration score.

Science.gov (United States)

Kendall, Katherine A

2017-10-01

Quantitative measures of swallowing function may improve the reliability and accuracy of modified barium swallow (MBS) study interpretation. Quantitative study analysis has not been widely instituted, however, secondary to concerns about the time required to make measures and a lack of research demonstrating impact on MBS interpretation. This study compares the accuracy of the penetration/aspiration (PEN/ASP) scale (an observational visual-perceptual assessment tool) to quantitative measures of airway closure timing relative to the arrival of the bolus at the upper esophageal sphincter in identifying a failure of airway protection during deglutition. Retrospective review of clinical swallowing data from a university-based outpatient clinic. Swallowing data from 426 patients were reviewed. Patients with normal PEN/ASP scores were identified, and the results of quantitative airway closure timing measures for three liquid bolus sizes were evaluated. The incidence of significant airway closure delay with and without a normal PEN/ASP score was determined. Inter-rater reliability for the quantitative measures was calculated. In patients with a normal PEN/ASP score, 33% demonstrated a delay in airway closure on at least one swallow during the MBS study. There was no correlation between PEN/ASP score and airway closure delay. Inter-rater reliability for the quantitative measure of airway closure timing was nearly perfect (intraclass correlation coefficient = 0.973). The use of quantitative measures of swallowing function, in conjunction with traditional visual perceptual methods of MBS study interpretation, improves the identification of airway closure delay, and hence, potential aspiration risk, even when no penetration or aspiration is apparent on the MBS study. 4. Laryngoscope, 127:2314-2318, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

Comparative Evaluation of Four Real-Time PCR Methods for the Quantitative Detection of Epstein-Barr Virus from Whole Blood Specimens.

Science.gov (United States)

Buelow, Daelynn; Sun, Yilun; Tang, Li; Gu, Zhengming; Pounds, Stanley; Hayden, Randall

2016-07-01

Monitoring of Epstein-Barr virus (EBV) load in immunocompromised patients has become integral to their care. An increasing number of reagents are available for quantitative detection of EBV; however, there are little published comparative data. Four real-time PCR systems (one using laboratory-developed reagents and three using analyte-specific reagents) were compared with one another for detection of EBV from whole blood. Whole blood specimens seeded with EBV were used to determine quantitative linearity, analytical measurement range, lower limit of detection, and CV for each assay. Retrospective testing of 198 clinical samples was performed in parallel with all methods; results were compared to determine relative quantitative and qualitative performance. All assays showed similar performance. No significant difference was found in limit of detection (3.12-3.49 log10 copies/mL; P = 0.37). A strong qualitative correlation was seen with all assays that used clinical samples (positive detection rates of 89.5%-95.8%). Quantitative correlation of clinical samples across assays was also seen in pairwise regression analysis, with R(2) ranging from 0.83 to 0.95. Normalizing clinical sample results to IU/mL did not alter the quantitative correlation between assays. Quantitative EBV detection by real-time PCR can be performed over a wide linear dynamic range, using three different commercially available reagents and laboratory-developed methods. EBV was detected with comparable sensitivity and quantitative correlation for all assays. Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Quantitative analysis of the dystrophin gene by real-time PCR

Directory of Open Access Journals (Sweden)

Maksimovic Nela

2012-01-01

Full Text Available Duchenne and Becker muscular dystrophy (DMD/BMD are severe X-linked neuromuscular disorders caused by mutations in the dystrophin gene. Our aim was to optimize a quantitative real-time PCR method based on SYBR® Green I chemistry for routine diagnostics of DMD/BMD deletion carriers. Twenty female relatives of DMD/BMD patients with previously detected partial gene deletions were studied. The relative quantity of the target exons was calculated by a comparative threshold cycle method (ΔΔCt. The carrier status of all subjects was successfully determined. The gene dosage ratio for non-carriers was 1.07±0.20, and for carriers 0.56±0.11. This assay proved to be simple, rapid, reliable and cost-effective.

Real-Time Quantitative PCR Measurement of Ileal Lactobacillus salivarius Populations from Broiler Chickens To Determine the Influence of Farming Practices▿ †

Science.gov (United States)

Harrow, Sally A.; Ravindran, Velmurugu; Butler, Ruth C.; Marshall, John W.; Tannock, Gerald W.

2007-01-01

A real-time quantitative PCR assay targeting a 16S-23S intergenic spacer region sequence was devised to measure the sizes of populations of Lactobacillus salivarius present in ileal digesta collected from broiler chickens. This species has been associated with deconjugation of bile salts in the small bowel and reduced broiler productivity. The assay was tested as a means of monitoring the sizes of L. salivarius populations from broilers fed diets with different compositions, maintained at different stocking densities, or given the antimicrobial drugs bacitracin and monensin in the feed. Stocking densities did not influence the numbers of L. salivarius cells in the ileum. A diet containing meat and bone meal reduced the size of the L. salivarius population relative to that of chickens given the control diet, as did administration of bacitracin and monensin in the feed. These changes in the target bacterial population were associated with improved broiler weight gain. PMID:17890342

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

Quantitative Real Time PCR approach to study gene expression profile during prenatal growth of skeletal muscle in pig of Duroc and Pietrain breeds

Directory of Open Access Journals (Sweden)

M. Cagnazzo

2010-01-01

Full Text Available The quantitative real time-PCR (QRT-PCR is a very sensitive method used to quantify mRNA level in gene expression analysis. Combining amplification, detection and quantification in a single step, allows a more accurate measurement compared to the traditional PCR end point analysis (Pfaffl, 2001; Bustin, 2002.

A real-time BWR stability measurement system

International Nuclear Information System (INIS)

March-Leuba, J.; King, W.T.

1988-01-01

This paper describes the characteristics of a portable, real-time system used for nonperturbational measurements of stability in boiling water reactors. The algorithm used in this system estimates the closed-loop asymptotic decay ratio using only the naturally occurring neutron noise and it is based on the univariate autoregressive methodology. (author)

[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.

Preliminary results of real-time in-vitro electronic speckle pattern interferometry (ESPI) measurements in otolaryngology

Science.gov (United States)

Conerty, Michelle D.; Castracane, James; Cacace, Anthony T.; Parnes, Steven M.; Gardner, Glendon M.; Miller, Mitchell B.

1995-05-01

Electronic Speckle Pattern Interferometry (ESPI) is a nondestructive optical evaluation technique that is capable of determining surface and subsurface integrity through the quantitative evaluation of static or vibratory motion. By utilizing state of the art developments in the areas of lasers, fiber optics and solid state detector technology, this technique has become applicable in medical research and diagnostics. Based on initial support from NIDCD and continued support from InterScience, Inc., we have been developing a range of instruments for improved diagnostic evaluation in otolaryngological applications based on the technique of ESPI. These compact fiber optic instruments are capable of making real time interferometric measurements of the target tissue. Ongoing development of image post- processing software is currently capable of extracting the desired quantitative results from the acquired interferometric images. The goal of the research is to develop a fully automated system in which the image processing and quantification will be performed in hardware in near real-time. Subsurface details of both the tympanic membrane and vocal cord dynamics could speed the diagnosis of otosclerosis, laryngeal tumors, and aid in the evaluation of surgical procedures.

Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR.

Science.gov (United States)

Furet, Jean-Pierre; Quénée, Pascal; Tailliez, Patrick

2004-12-15

Real-time quantitative PCR assays were developed for the absolute quantification of lactic acid bacteria (LAB) (Streptococcus thermophilus, Lactobacillus delbrueckii, L. casei, L. paracasei, L. rhamnosus, L. acidophilus and L. johnsonii) in fermented milk products. The results of molecular quantification and classic bacterial enumeration did not differ significantly with respect to S. thermophilus and the species of the L. casei group which were detected in the six commercial fermented products tested, thus showing that DNA extraction was efficient and that genomic DNA solutions were free of PCR inhibitors. For L. delbrueckii, the results of bacterial enumeration were generally lower by a factor 10 to 100 than those of PCR quantification, suggesting a loss of viability during storage of the dairy products at 1-8 degrees C for most of the strains in this species. Real-time quantitative assays enabled identification of the species of lactic acid bacterial strains initially present in commercial fermented milk products and their accurate quantification with a detection threshold of 10(3) cells per ml of product.

Grasp: Tracing, visualizing and measuring the behavior of real-time systems

NARCIS (Netherlands)

Holenderski, M.J.; Heuvel, van den M.M.H.P.; Bril, R.J.; Lukkien, J.J.; Lipari, G.; Cucinotta, T.

2010-01-01

Understanding and validating the timing behavior of real-time systems is not trivial. Many real-time operating systems and their development environments do not provide tracing support, and provide only limited visualization, measurements and analysis tools. This paper presents Grasp, a tool for

Quantitative detection of Campylobacter jejuni on fresh chicken carcasses by real-time PCR.

Science.gov (United States)

Rönner, Anna-Clara; Lindmark, Hans

2007-06-01

Campylobacter jejuni infection is a significant cause of foodborne gastroenteritis worldwide. Consumption and handling of poultry products is believed to be the primary risk factor for campylobacteriosis. Risk assessments require quantitative data, and C. jejuni is enumerated usually by direct plating, which sometimes allows growth of non-Campylobacter bacteria. The objective of the present study was to develop a quantitative real-time PCR method (q-PCR) for enumerating C. jejuni in chicken rinse without a culturing step. The procedure to obtain the template for the PCR assay involved (i) filtration of 10 ml of chicken rinse, (ii) centrifugation of the sample, and (iii) total DNA extraction from the pellet obtained using a commercial DNA extraction kit. The detection limit of the method was comparable to that for plating 100 microl of chicken rinse on modified charcoal cefoperazone deoxycholate agar, and the detection limit could be further improved 10-fold by concentrating the DNA eluate by ethanol precipitation. A close correlation for spiked chicken rinse was obtained for the results of the quantitative real-time PCR method and direct plating (r = 0.99). The coefficient of correlation for the methods was 0.87 when samples from chicken carcasses on the slaughter line were analyzed, whereas a lower correlation (r = 0.76) was obtained when samples from retail carcasses were analyzed. Greater variation in the proportion of dead and/or viable but not culturable Campylobacter types in the retail samples may explain the decreased correlation between the methods. Overall, the new method is simple and fast and the results obtained are closely correlated with those for direct plating for samples containing a low proportion of dead Campylobacter cells.

Interlaboratory validation of quantitative duplex real-time PCR method for screening analysis of genetically modified maize.

Science.gov (United States)

Takabatake, Reona; Koiwa, Tomohiro; Kasahara, Masaki; Takashima, Kaori; Futo, Satoshi; Minegishi, Yasutaka; Akiyama, Hiroshi; Teshima, Reiko; Oguchi, Taichi; Mano, Junichi; Furui, Satoshi; Kitta, Kazumi

2011-01-01

To reduce the cost and time required to routinely perform the genetically modified organism (GMO) test, we developed a duplex quantitative real-time PCR method for a screening analysis simultaneously targeting an event-specific segment for GA21 and Cauliflower Mosaic Virus 35S promoter (P35S) segment [Oguchi et al., J. Food Hyg. Soc. Japan, 50, 117-125 (2009)]. To confirm the validity of the method, an interlaboratory collaborative study was conducted. In the collaborative study, conversion factors (Cfs), which are required to calculate the GMO amount (%), were first determined for two real-time PCR instruments, the ABI PRISM 7900HT and the ABI PRISM 7500. A blind test was then conducted. The limit of quantitation for both GA21 and P35S was estimated to be 0.5% or less. The trueness and precision were evaluated as the bias and reproducibility of the relative standard deviation (RSD(R)). The determined bias and RSD(R) were each less than 25%. We believe the developed method would be useful for the practical screening analysis of GM maize.

Quantitative assessment of hematopoietic chimerism by quantitative real-time polymerase chain reaction of sequence polymorphism systems after hematopoietic stem cell transplantation.

Science.gov (United States)

Qin, Xiao-ying; Li, Guo-xuan; Qin, Ya-zhen; Wang, Yu; Wang, Feng-rong; Liu, Dai-hong; Xu, Lan-ping; Chen, Huan; Han, Wei; Wang, Jing-zhi; Zhang, Xiao-hui; Li, Jin-lan; Li, Ling-di; Liu, Kai-yan; Huang, Xiao-jun

2011-08-01

Analysis of changes in recipient and donor hematopoietic cell origin is extremely useful to monitor the effect of hematopoietic stem cell transplantation (HSCT) and sequential adoptive immunotherapy by donor lymphocyte infusions. We developed a sensitive, reliable and rapid real-time PCR method based on sequence polymorphism systems to quantitatively assess the hematopoietic chimerism after HSCT. A panel of 29 selected sequence polymorphism (SP) markers was screened by real-time PCR in 101 HSCT patients with leukemia and other hematological diseases. The chimerism kinetics of bone marrow samples of 8 HSCT patients in remission and relapse situations were followed longitudinally. Recipient genotype discrimination was possible in 97.0% (98 of 101) with a mean number of 2.5 (1-7) informative markers per recipient/donor pair. Using serial dilutions of plasmids containing specific SP markers, the linear correlation (r) of 0.99, the slope between -3.2 and -3.7 and the sensitivity of 0.1% were proved reproducible. By this method, it was possible to very accurately detect autologous signals in the range from 0.1% to 30%. The accuracy of the method in the very important range of autologous signals below 5% was extraordinarily high (standard deviation real-time PCR method over short tandem repeat PCR chimerism assays is the absence of PCR competition and plateau biases, with demonstrated greater sensitivity and linearity. Finally, we prospectively analyzed bone marrow samples of 8 patients who received allografts and presented the chimerism kinetics of remission and relapse situations that illustrated the sensitivity level and the promising clinical application of this method. This SP-based real-time PCR assay provides a rapid, sensitive, and accurate quantitative assessment of mixed chimerism that can be useful in predicting graft rejection and early relapse.

Decay Of Bacterial Pathogens, Fecal Indicators, And Real-Time Quantitative PCR Genetic Markers In Manure-Amended Soils

Science.gov (United States)

This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manure-amended agricultural soils. Known concentrations of transformed green...

Decay Of Bacterial Pathogen, Fecal Indicators, And Real-Time Quantitative PCR Genetic Markers In Manure Amended Soils

Science.gov (United States)

This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria, and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manre-amended agricultural soils. Known concentrations of transformed green fluore...

Real-Time Intracellular Measurements of ROS and RNS in Living Cells with Single Core-Shell Nanowire Electrodes.

Science.gov (United States)

Zhang, Xin-Wei; Qiu, Quan-Fa; Jiang, Hong; Zhang, Fu-Li; Liu, Yan-Lin; Amatore, Christian; Huang, Wei-Hua

2017-10-09

Nanoelectrodes allow precise and quantitative measurements of important biological processes at the single living-cell level in real time. Cylindrical nanowire electrodes (NWEs) required for intracellular measurements create a great challenge for achieving excellent electrochemical and mechanical performances. Herein, we present a facile and robust solution to this problem based on a unique SiC-core-shell design to produce cylindrical NWEs with superior mechanical toughness provided by the SiC nano-core and an excellent electrochemical performance provided by the ultrathin carbon shell that can be used as such or platinized. The use of such NWEs for biological applications is illustrated by the first quantitative measurements of ROS/RNS in individual phagolysosomes of living macrophages. As the shell material can be varied to meet any specific detection purpose, this work opens up new opportunities to monitor quantitatively biological functions occurring inside cells and their organelles. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Real-time database for high resolution neutron monitor measurements

Energy Technology Data Exchange (ETDEWEB)

Steigies, Christian T.; Rother, Oliver M.; Wimmer-Schweingruber, Robert F.; Heber, Bernd [IEAP, Christian-Albrechts-Universitaet zu Kiel (Germany)

2008-07-01

The worldwide network of standardised neutron monitors is, after 50 years, still the state-of-the-art instrumentation to measure spectral variations of the primary cosmic ray component. These measurements are an ideal complement to space based cosmic ray measurements. Data from the approximately 50 IGY and NM64 neutron monitors is stored locally but also available through data collections sites like the World Data Center (WDC) or the IZMIRAN ftp server. The data from the WDC is in a standard format, but only hourly values are available. IZMIRAN collects the data in the best available time resolution, but the data arrives on the ftp server only hours, sometimes days, after the measurements. Also, the high time-resolution measurements of the different stations do not have a common format, a conversion routine for each station is needed before they can be used for scientific analysis. Supported by the 7th framework program of the European Commission, we are setting up a real-time database where high resolution cosmic ray measurements will be stored and accessible immediately after the measurement. Stations that do not have 1-minute resolution measurements will be upgraded to 1-minute or better resolution with an affordable standard registration system, that will submit the measurements to the database via the internet in real-time.

EVALUATION OF A RAPID, QUANTITATIVE REAL-TIME PCR METHOD FOR ENUMERATION OF PATHOGENIC CANDIDA CELLS IN WATER

Science.gov (United States)

Quantitative Real-Time PCR (QRT-PCR) technology, incorporating fluorigenic 5' nuclease (TaqMan?) chemistry, was developed for the specific detection and quantification of six pathogenic species of Candida (C. albicans, C. tropicalis, C. krusei, C. parapsilosis, C. glabrata and C....

Comparison of Hybrid Capture 2 Assay with Real-time-PCR for Detection and Quantitation of Hepatitis B Virus DNA.

Science.gov (United States)

Majid, Farjana; Jahan, Munira; Lutful Moben, Ahmed; Tabassum, Shahina

2014-01-01

Both real-time-polymerase chain reaction (PCR) and hybrid capture 2 (HC2) assay can detect and quantify hepatitis B virus (HBV) DNA. However, real-time-PCR can detect a wide range of HBV DNA, while HC2 assay could not detect lower levels of viremia. The present study was designed to detect and quantify HBV DNA by real-time-PCR and HC2 assay and compare the quantitative data of these two assays. A cross-sectional study was conducted in between July 2010 and June 2011. A total of 66 serologically diagnosed chronic hepatitis B (CHB) patients were selected for the study. Real-time-PCR and HC2 assay was done to detect HBV DNA. Data were analyzed by statistical Package for the social sciences (SPSS). Among 66 serologically diagnosed chronic hepatitis B patients 40 (60.61%) patients had detectable and 26 (39.39%) had undetectable HBV DNA by HC2 assay. Concordant results were obtained for 40 (60.61%) out of these 66 patients by real-time-PCR and HC2 assay with mean viral load of 7.06 ± 1.13 log 10 copies/ml and 6.95 ± 1.08 log 10 copies/ml, respectively. In the remaining 26 patients, HBV DNA was detectable by real-time-PCR in 20 patients (mean HBV DNA level was 3.67 ± 0.72 log 10 copies/ml. However, HBV DNA could not be detectable in six cases by the both assays. The study showed strong correlation (r = 0.915) between real-time-PCR and HC2 assay for the detection and quantification of HBV DNA. HC2 assay may be used as an alternative to real-time-PCR for CHB patients. How to cite this article: Majid F, Jahan M, Moben AL, Tabassum S. Comparison of Hybrid Capture 2 Assay with Real-time-PCR for Detection and Quantitation of Hepatitis B Virus DNA. Euroasian J Hepato-Gastroenterol 2014;4(1):31-35.

Real-Time 3D Profile Measurement Using Structured Light

International Nuclear Information System (INIS)

Xu, L; Zhang, Z J; Ma, H; Yu, Y J

2006-01-01

The paper builds a real-time system of 3D profile measurement using structured-light imaging. It allows a hand-held object to rotate free in the space-time coded light field, which is projected by the projector. The surface of measured objects with projected coded light is imaged; the system shows surface reconstruction results of objects online. This feedback helps user to adjust object's pose in the light field according to the dismissed or error data, which would achieve the integrality of data used in reconstruction. This method can acquire denser data cloud and have higher reconstruction accuracy and efficiency. According to the real-time requirements, the paper presents the non-restricted light plane modelling which suits stripe structured light system, designs the three-frame stripes space-time coded pattern, and uses the advance ICP algorithms to acquire 3D data alignment from multiple view

Development of a Real-Time Resistance Measurement for Vibrio parahaemolyticus Detection by the Lecithin-Dependent Hemolysin Gene

Science.gov (United States)

Xiang, Guiming; Pu, Xiaoyun; Jiang, Dongneng; Liu, Linlin; Liu, Chang; Liu, Xiaobo

2013-01-01

The marine bacterium Vibrio parahaemolyticus (V. parahaemolyticus) causes gastroenteritis in humans via the ingestion of raw or undercooked contaminated seafood, and early diagnosis and prompt treatment are important for the prevention of V. parahaemolyticus-related diseases. In this study, a real-time resistance measurement based on loop-mediated isothermal amplification (LAMP), electrochemical ion bonding (Crystal violet and Mg2+), real-time monitoring, and derivative analysis was developed. V. parahaemolyticus DNA was first amplified by LAMP, and the products (DNA and pyrophosphate) represented two types of negative ions that could combine with a positive dye (Crystal violet) and positive ions (Mg2+) to increase the resistance of the reaction liquid. This resistance was measured in real-time using a specially designed resistance electrode, thus permitting the quantitative detection of V. parahaemolyticus. The results were obtained in 1–2 hours, with a minimum bacterial density of 10 CFU.mL−1 and high levels of accuracy (97%), sensitivity (96.08%), and specificity (97.96%) when compared to cultivation methods. Therefore, this simple and rapid method has a potential application in the detection of V. parahaemolyticus on a gene chip or in point-of-care testing. PMID:23991096

Real-time evolvable pulse shaper for radiation measurements

Energy Technology Data Exchange (ETDEWEB)

Lanchares, Juan, E-mail: julandan@dacya.ucm.es [Facultad de Informática, Universidad Complutense de Madrid (UCM), C/Prof. José García Santesmases s/n, 28040 Madrid (Spain); Garnica, Oscar, E-mail: ogarnica@dacya.ucm.es [Facultad de Informática, Universidad Complutense de Madrid (UCM), C/Prof. José García Santesmases s/n, 28040 Madrid (Spain); Risco-Martín, José L., E-mail: jlrisco@dacya.ucm.es [Facultad de Informática, Universidad Complutense de Madrid (UCM), C/Prof. José García Santesmases s/n, 28040 Madrid (Spain); Ignacio Hidalgo, J., E-mail: hidalgo@dacya.ucm.es [Facultad de Informática, Universidad Complutense de Madrid (UCM), C/Prof. José García Santesmases s/n, 28040 Madrid (Spain); Regadío, Alberto, E-mail: alberto.regadio@insa.es [Área de Tecnologías Electrónicas, Instituto Nacional de Técnica Aeroespacial (INTA), 28850 Torrejón de Ardoz, Madrid (Spain)

2013-11-01

In the last two decades, recursive algorithms for real-time digital pulse shaping in pulse height measurements have been developed and published in number of articles and textbooks. All these algorithms try to synthesize in real time optimum or near optimum shapes in the presence of noise. Even though some of these shapers can be considered effective designs, some side effects like aging cannot be ignored. We may observe that after sensors degradation, the signal obtained is not valid. In this regard, we present in this paper a novel technique that, based on evolvable hardware concepts, is able to evolve the degenerated shaper into a new design with better performance than the original one under the new sensor features.

Real-time measurement of relative sensor position changes using ultrasonic signal evaluation

Energy Technology Data Exchange (ETDEWEB)

Yastrebova, O.; Bulavinov, A.; Kroening, M. [Fraunhofer Institute Nondestructive Testing IZFP, Saarbruecken (Germany)

2008-07-01

Ultrasonic testing is considered to be one of the most commonly applied nondestructive testing techniques for flaw detection and material characterization. Traditional Nondestructive Testing (NDT) provides detection of material discontinuities that may cause failure within the designed lifetime of a part or component. In addition, Quantitative Nondestructive Testing (QNDT) provides means to obtain required information about type, size and location of deficiencies to the integrity of the inspected structure and further use under specific, given load conditions. The ''Acoustic Mouse'' technique has been developed as a tool for manual ultrasonic inspection to provide test results that can be evaluated quantitatively. The ultrasonic data are processed by real-time variation methods to extract position information from backscattered acoustic noise and geometric scatter signals in the inspection volume. The position and positional changes of the ''Acoustic Mouse'' sensor (transducer) are determined by the sequential analysis of ultrasonic data (highresolution sector-scans), which are acquired and reconstructed using the Sampling Phased Array technique. The results of first experiments conducted with linear scanning and intentional lift-offs demonstrate sufficient accuracy in position measurements. (orig.)

Rapid quantitative detection of Lactobacillus sakei in meat and fermented sausages by real-time PCR.

Science.gov (United States)

Martín, Belén; Jofré, Anna; Garriga, Margarita; Pla, Maria; Aymerich, Teresa

2006-09-01

A quick and simple method for quantitative detection of Lactobacillus sakei in fermented sausages was successfully developed. It is based on Chelex-100-based DNA purification and real-time PCR enumeration using a TaqMan fluorescence probe. Primers and probes were designed in the L. sakei 16S-23S rRNA intergenic transcribed spacer region, and the assay was evaluated using L. sakei genomic DNA and an artificially inoculated sausage model. The detection limit of this technique was approximately 3 cells per reaction mixture using both purified DNA and the inoculated sausage model. The quantification limit was established at 30 cells per reaction mixture in both models. The assay was then applied to enumerate L. sakei in real samples, and the results were compared to the MRS agar count method followed by confirmation of the percentage of L. sakei colonies. The results obtained by real-time PCR were not statistically significantly different than those obtained by plate count on MRS agar (P > 0.05), showing a satisfactory agreement between both methods. Therefore, the real-time PCR assay developed can be considered a promising rapid alternative method for the quantification of L. sakei and evaluation of the implantation of starter strains of L. sakei in fermented sausages.

Legionellosis and Lung Abscesses: Contribution of Legionella Quantitative Real-Time PCR to an Adapted Followup

Directory of Open Access Journals (Sweden)

G. Descours

2013-01-01

Full Text Available We report a case of severe Legionnaires' disease (LD complicated by a lung abscess in an immunocompetent patient who required ECMO therapy and thoracic surgery. The results of repeated Legionella quantitative real-time PCR performed on both sera and respiratory samples correlated with the LD severity and the poor clinical outcome. Moreover, the PCR allowed for the detection of Legionella DNA in the lung abscess specimen, which was negative when cultured for Legionella. This case report provides a logical basis for further investigations to examine whether the Legionella quantitative PCR could improve the assessment of LD severity and constitute a prognostic marker.

Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process

Directory of Open Access Journals (Sweden)

Borges-Pérez Andrés

2008-12-01

Full Text Available Abstract Background The elucidation of gene expression patterns leads to a better understanding of biological processes. Real-time quantitative RT-PCR has become the standard method for in-depth studies of gene expression. A biologically meaningful reporting of target mRNA quantities requires accurate and reliable normalization in order to identify real gene-specific variation. The purpose of normalization is to control several variables such as different amounts and quality of starting material, variable enzymatic efficiencies of retrotranscription from RNA to cDNA, or differences between tissues or cells in overall transcriptional activity. The validity of a housekeeping gene as endogenous control relies on the stability of its expression level across the sample panel being analysed. In the present report we describe the first systematic evaluation of potential internal controls during tomato development process to identify which are the most reliable for transcript quantification by real-time RT-PCR. Results In this study, we assess the expression stability of 7 traditional and 4 novel housekeeping genes in a set of 27 samples representing different tissues and organs of tomato plants at different developmental stages. First, we designed, tested and optimized amplification primers for real-time RT-PCR. Then, expression data from each candidate gene were evaluated with three complementary approaches based on different statistical procedures. Our analysis suggests that SGN-U314153 (CAC, SGN-U321250 (TIP41, SGN-U346908 ("Expressed" and SGN-U316474 (SAND genes provide superior transcript normalization in tomato development studies. We recommend different combinations of these exceptionally stable housekeeping genes for suited normalization of different developmental series, including the complete tomato development process. Conclusion This work constitutes the first effort for the selection of optimal endogenous controls for quantitative real-time

Application of quantitative real-time PCR compared to filtration methods for the enumeration of Escherichia coli in surface waters within Vietnam.

Science.gov (United States)

Vital, Pierangeli G; Van Ha, Nguyen Thi; Tuyet, Le Thi Hong; Widmer, Kenneth W

2017-02-01

Surface water samples in Vietnam were collected from the Saigon River, rural and suburban canals, and urban runoff canals in Ho Chi Minh City, Vietnam, and were processed to enumerate Escherichia coli. Quantification was done through membrane filtration and quantitative real-time polymerase chain reaction (PCR). Mean log colony-forming unit (CFU)/100 ml E. coli counts in the dry season for river/suburban canals and urban canals were log 2.8 and 3.7, respectively, using a membrane filtration method, while using Taqman quantitative real-time PCR they were log 2.4 and 2.8 for river/suburban canals and urban canals, respectively. For the wet season, data determined by the membrane filtration method in river/suburban canals and urban canals samples had mean counts of log 3.7 and 4.1, respectively. While mean log CFU/100 ml counts in the wet season using quantitative PCR were log 3 and 2, respectively. Additionally, the urban canal samples were significantly lower than those determined by conventional culture methods for the wet season. These results show that while quantitative real-time PCR can be used to determine levels of fecal indicator bacteria in surface waters, there are some limitations to its application and it may be impacted by sources of runoff based on surveyed samples.

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.

Identification of circulating miRNA biomarkers based on global quantitative real-time PCR profiling

Directory of Open Access Journals (Sweden)

Kang Kang

2012-02-01

Full Text Available Abstract MicroRNAs (miRNAs are small noncoding RNAs (18-25 nucleotides that regulate gene expression at the post-transcriptional level. Recent studies have demonstrated the presence of miRNAs in the blood circulation. Deregulation of miRNAs in serum or plasma has been associated with many diseases including cancers and cardiovascular diseases, suggesting the possible use of miRNAs as diagnostic biomarkers. However, the detection of the small amount of miRNAs found in serum or plasma requires a method with high sensitivity and accuracy. Therefore, the current study describes polymerase chain reaction (PCR-based methods for measuring circulating miRNAs. Briefly, the procedure involves four major steps: (1 sample collection and preparation; (2 global miRNAs profiling using quantitative real-time PCR (qRT-PCR; (3 data normalization and analysis; and (4 selection and validation of miRNA biomarkers. In conclusion, qRT-PCR is a promising method for profiling of circulating miRNAs as biomarkers.

Quantitation of hepatitis B virus DNA in plasma using a sensitive cost-effective "in-house" real-time PCR assay

Directory of Open Access Journals (Sweden)

Daniel Hubert Darius

2009-01-01

Full Text Available Background: Sensitive nucleic acid testing for the detection and accurate quantitation of hepatitis B virus (HBV is necessary to reduce transmission through blood and blood products and for monitoring patients on antiviral therapy. The aim of this study is to standardize an "in-house" real-time HBV polymerase chain reaction (PCR for accurate quantitation and screening of HBV. Materials and Methods: The "in-house" real-time assay was compared with a commercial assay using 30 chronically infected individuals and 70 blood donors who are negative for hepatitis B surface antigen, hepatitis C virus (HCV antibody and human immunodeficiency virus (HIV antibody. Further, 30 HBV-genotyped samples were tested to evaluate the "in-house" assay′s capacity to detect genotypes prevalent among individuals attending this tertiary care hospital. Results: The lower limit of detection of this "in-house" HBV real-time PCR was assessed against the WHO international standard and found to be 50 IU/mL. The interassay and intra-assay coefficient of variation (CV of this "in-house" assay ranged from 1.4% to 9.4% and 0.0% to 2.3%, respectively. Virus loads as estimated with this "in-house" HBV real-time assay correlated well with the commercial artus HBV RG PCR assay ( r = 0.95, P < 0.0001. Conclusion: This assay can be used for the detection and accurate quantitation of HBV viral loads in plasma samples. This assay can be employed for the screening of blood donations and can potentially be adapted to a multiplex format for simultaneous detection of HBV, HIV and HCV to reduce the cost of testing in blood banks.

Real time operation of a multiple gamma measurement installation

International Nuclear Information System (INIS)

Philippot, J.C.; Lefevre, J.

1980-01-01

This paper describes a multiple measurement channel facility for fine gamma spectrometry, its real time operation, and the new possibilities which it offers. The installation is presented in its twofold electronic and processing aspects, by considering its architecture, its hard and software, and its data processing package. Real time operation requires customized general organization, perfect instantaneous knowledge of the status of all the units, and a sound hierarchy between the various participants, operators as well as requestors. The care inherent in the installation itself and in the definition of its operation explains its new possibilities. (Auth.)

Real Time Speed Measure while Automobile Braking on Soft Sensing Technique

Energy Technology Data Exchange (ETDEWEB)

Zhu, W B; Li, D S; Lu, Y [China Jiliang University, Hangzhou, Zhejiang province, 310018 (China)

2006-10-15

Because the braking performance of automobile has close relationship to traffic safety, it is important to detect that. Focusing on the problem that the real time speed is difficult to obtain in detection process, soft sensing technique is introduced in this paper. According to analyzing the relationship of the dynamics equation of a moving automobile, a module of real time speed of braking is set up. By using imitation method with experiment data to get the pressure function of cylinder and analyzing the relationship between the trigging moment of a wheel and the pressure function of brake cylinder, the real time speed is confirmed in good precision. The maximal measurement error of real time speed is 8.7% and the precision can satisfy engineering request.

Real Time Speed Measure while Automobile Braking on Soft Sensing Technique

International Nuclear Information System (INIS)

Zhu, W B; Li, D S; Lu, Y

2006-01-01

Because the braking performance of automobile has close relationship to traffic safety, it is important to detect that. Focusing on the problem that the real time speed is difficult to obtain in detection process, soft sensing technique is introduced in this paper. According to analyzing the relationship of the dynamics equation of a moving automobile, a module of real time speed of braking is set up. By using imitation method with experiment data to get the pressure function of cylinder and analyzing the relationship between the trigging moment of a wheel and the pressure function of brake cylinder, the real time speed is confirmed in good precision. The maximal measurement error of real time speed is 8.7% and the precision can satisfy engineering request

Development and evaluation of a real-time one step Reverse-Transcriptase PCR for quantitation of Chandipura Virus

Directory of Open Access Journals (Sweden)

Tandale Babasaheb V

2008-12-01

Full Text Available Abstract Background Chandipura virus (CHPV, a member of family Rhabdoviridae was attributed to an explosive outbreak of acute encephalitis in children in Andhra Pradesh, India in 2003 and a small outbreak among tribal children from Gujarat, Western India in 2004. The case-fatality rate ranged from 55–75%. Considering the rapid progression of the disease and high mortality, a highly sensitive method for quantifying CHPV RNA by real-time one step reverse transcriptase PCR (real-time one step RT-PCR using TaqMan technology was developed for rapid diagnosis. Methods Primers and probe for P gene were designed and used to standardize real-time one step RT-PCR assay for CHPV RNA quantitation. Standard RNA was prepared by PCR amplification, TA cloning and run off transcription. The optimized real-time one step RT-PCR assay was compared with the diagnostic nested RT-PCR and different virus isolation systems [in vivo (mice in ovo (eggs, in vitro (Vero E6, PS, RD and Sand fly cell line] for the detection of CHPV. Sensitivity and specificity of real-time one step RT-PCR assay was evaluated with diagnostic nested RT-PCR, which is considered as a gold standard. Results Real-time one step RT-PCR was optimized using in vitro transcribed (IVT RNA. Standard curve showed linear relationship for wide range of 102-1010 (r2 = 0.99 with maximum Coefficient of variation (CV = 5.91% for IVT RNA. The newly developed real-time RT-PCR was at par with nested RT-PCR in sensitivity and superior to cell lines and other living systems (embryonated eggs and infant mice used for the isolation of the virus. Detection limit of real-time one step RT-PCR and nested RT-PCR was found to be 1.2 × 100 PFU/ml. RD cells, sand fly cells, infant mice, and embryonated eggs showed almost equal sensitivity (1.2 × 102 PFU/ml. Vero and PS cell-lines (1.2 × 103 PFU/ml were least sensitive to CHPV infection. Specificity of the assay was found to be 100% when RNA from other viruses or healthy

Real-Time Video Stylization Using Object Flows.

Science.gov (United States)

Lu, Cewu; Xiao, Yao; Tang, Chi-Keung

2017-05-05

We present a real-time video stylization system and demonstrate a variety of painterly styles rendered on real video inputs. The key technical contribution lies on the object flow, which is robust to inaccurate optical flow, unknown object transformation and partial occlusion as well. Since object flows relate regions of the same object across frames, shower-door effect can be effectively reduced where painterly strokes and textures are rendered on video objects. The construction of object flows is performed in real time and automatically after applying metric learning. To reduce temporal flickering, we extend the bilateral filtering into motion bilateral filtering. We propose quantitative metrics to measure the temporal coherence on structures and textures of our stylized videos, and perform extensive experiments to compare our stylized results with baseline systems and prior works specializing in watercolor and abstraction.

Improved process control through real-time measurement of mineral content

Energy Technology Data Exchange (ETDEWEB)

Turler, Daniel; Karaca, Murat; Davis, William B.; Giauque, Robert D.; Hopkins, Deborah

2001-11-02

In a highly collaborative research and development project with mining and university partners, sensors and data-analysis tools are being developed for rock-mass characterization and real-time measurement of mineral content. Determining mineralogy prior to mucking in an open-pit mine is important for routing the material to the appropriate processing stream. A possible alternative to lab assay of dust and cuttings obtained from drill holes is continuous on-line sampling and real-time x-ray fluorescence (XRF) spectroscopy. Results presented demonstrate that statistical analyses combined with XRF data can be employed to identify minerals and, possibly, different rock types. The objective is to create a detailed three-dimensional mineralogical map in real time that would improve downstream process efficiency.

A human fecal contamination index for ranking impaired recreational watersusing the HF183 quantitative real-time PCR method

Science.gov (United States)

Human fecal pollution of surface water remains a public health concern worldwide. As a result, there is a growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for recreational water quality risk managem...

Quantitative real-time monitoring of multi-elements in airborne particulates by direct introduction into an inductively coupled plasma mass spectrometer

International Nuclear Information System (INIS)

Suzuki, Yoshinari; Sato, Hikaru; Hiyoshi, Katsuhiro; Furuta, Naoki

2012-01-01

A new calibration system for real-time determination of trace elements in airborne particulates was developed. Airborne particulates were directly introduced into an inductively coupled plasma mass spectrometer, and the concentrations of 15 trace elements were determined by means of an external calibration method. External standard solutions were nebulized by an ultrasonic nebulizer (USN) coupled with a desolvation system, and the resulting aerosol was introduced into the plasma. The efficiency of sample introduction via the USN was calculated by two methods: (1) the introduction of a Cr standard solution via the USN was compared with introduction of a Cr(CO) 6 standard gas via a standard gas generator and (2) the aerosol generated by the USN was trapped on filters and then analyzed. The Cr introduction efficiencies obtained by the two methods were the same, and the introduction efficiencies of the other elements were equal to the introduction efficiency of Cr. Our results indicated that our calibration method for introduction efficiency worked well for the 15 elements (Ti, V, Cr, Mn, Co, Ni, Cu, Zn, As, Mo, Sn, Sb, Ba, Tl and Pb). The real-time data and the filter-collection data agreed well for elements with low-melting oxides (V, Co, As, Mo, Sb, Tl, and Pb). In contrast, the real-time data were smaller than the filter-collection data for elements with high-melting oxides (Ti, Cr, Mn, Ni, Cu, Zn, Sn, and Ba). This result implies that the oxides of these 8 elements were not completely fused, vaporized, atomized, and ionized in the initial radiation zone of the inductively coupled plasma. However, quantitative real-time monitoring can be realized after correction for the element recoveries which can be calculated from the ratio of real-time data/filter-collection data. - Highlights: ► APs were directly introduced into ICP-MS and real-time analysis was performed. ► The real-time data were calibrated by a multi-element standard solution from USN. ► During real-time

Real-time electron density measurements from Cotton-Mouton effect in JET machine

International Nuclear Information System (INIS)

Brombin, M.; Boboc, A.; Zabeo, L.; Murari, A.

2008-01-01

Real-time density profile measurements are essential for advanced fusion tokamak operation and interferometry is a proven method for this task. Nevertheless, as a consequence of edge localized modes, pellet injections, fast density increases, or disruptions, the interferometer is subject to fringe jumps, which produce loss of the signal preventing reliable use of the measured density in a real-time feedback controller. An alternative method to measure the density is polarimetry based on the Cotton-Mouton effect, which is proportional to the line-integrated electron density. A new analysis approach has been implemented and tested to verify the reliability of the Cotton-Mouton measurements for a wide range of plasma parameters and to compare the density evaluated from polarimetry with that from interferometry. The density measurements based on polarimetry are going to be integrated in the real-time control system of JET since the difference with the interferometry is within one fringe for more than 90% of the cases.

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…

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)

Alchemy: A web 2.0 real-time quality assurance platform for human immunodeficiency Virus, hepatitis C Virus, and BK Virus quantitation assays

Directory of Open Access Journals (Sweden)

Emmanuel Agosto-Arroyo

2017-01-01

Full Text Available Background: The molecular diagnostics laboratory faces the challenge of improving test turnaround time (TAT. Low and consistent TATs are of great clinical and regulatory importance, especially for molecular virology tests. Laboratory information systems (LISs contain all the data elements necessary to do accurate quality assurance (QA reporting of TAT and other measures, but these reports are in most cases still performed manually: a time-consuming and error-prone task. The aim of this study was to develop a web-based real-time QA platform that would automate QA reporting in the molecular diagnostics laboratory at our institution, and minimize the time expended in preparing these reports. Methods: Using a standard Linux, Nginx, MariaDB, PHP stack virtual machine running atop a Dell Precision 5810, we designed and built a web-based QA platform, code-named Alchemy. Data files pulled periodically from the LIS in comma-separated value format were used to autogenerate QA reports for the human immunodeficiency virus (HIV quantitation, hepatitis C virus (HCV quantitation, and BK virus (BKV quantitation. Alchemy allowed the user to select a specific timeframe to be analyzed and calculated key QA statistics in real-time, including the average TAT in days, tests falling outside the expected TAT ranges, and test result ranges. Results: Before implementing Alchemy, reporting QA for the HIV, HCV, and BKV quantitation assays took 45–60 min of personnel time per test every month. With Alchemy, that time has decreased to 15 min total per month. Alchemy allowed the user to select specific periods of time and analyzed the TAT data in-depth without the need of extensive manual calculations. Conclusions: Alchemy has significantly decreased the time and the human error associated with QA report generation in our molecular diagnostics laboratory. Other tests will be added to this web-based platform in future updates. This effort shows the utility of informatician

Alchemy: A Web 2.0 Real-time Quality Assurance Platform for Human Immunodeficiency Virus, Hepatitis C Virus, and BK Virus Quantitation Assays.

Science.gov (United States)

Agosto-Arroyo, Emmanuel; Coshatt, Gina M; Winokur, Thomas S; Harada, Shuko; Park, Seung L

2017-01-01

The molecular diagnostics laboratory faces the challenge of improving test turnaround time (TAT). Low and consistent TATs are of great clinical and regulatory importance, especially for molecular virology tests. Laboratory information systems (LISs) contain all the data elements necessary to do accurate quality assurance (QA) reporting of TAT and other measures, but these reports are in most cases still performed manually: a time-consuming and error-prone task. The aim of this study was to develop a web-based real-time QA platform that would automate QA reporting in the molecular diagnostics laboratory at our institution, and minimize the time expended in preparing these reports. Using a standard Linux, Nginx, MariaDB, PHP stack virtual machine running atop a Dell Precision 5810, we designed and built a web-based QA platform, code-named Alchemy. Data files pulled periodically from the LIS in comma-separated value format were used to autogenerate QA reports for the human immunodeficiency virus (HIV) quantitation, hepatitis C virus (HCV) quantitation, and BK virus (BKV) quantitation. Alchemy allowed the user to select a specific timeframe to be analyzed and calculated key QA statistics in real-time, including the average TAT in days, tests falling outside the expected TAT ranges, and test result ranges. Before implementing Alchemy, reporting QA for the HIV, HCV, and BKV quantitation assays took 45-60 min of personnel time per test every month. With Alchemy, that time has decreased to 15 min total per month. Alchemy allowed the user to select specific periods of time and analyzed the TAT data in-depth without the need of extensive manual calculations. Alchemy has significantly decreased the time and the human error associated with QA report generation in our molecular diagnostics laboratory. Other tests will be added to this web-based platform in future updates. This effort shows the utility of informatician-supervised resident/fellow programming projects as learning

A multiplex calibrated real-time PCR assay for quantitation of DNA of EBV-1 and 2.

Science.gov (United States)

Gatto, Francesca; Cassina, Giulia; Broccolo, Francesco; Morreale, Giuseppe; Lanino, Edoardo; Di Marco, Eddi; Vardas, Efthiya; Bernasconi, Daniela; Buttò, Stefano; Principi, Nicola; Esposito, Susanna; Scarlatti, Gabriella; Lusso, Paolo; Malnati, Mauro S

2011-12-01

Accurate and highly sensitive tests for the diagnosis of active Epstein-Barr virus (EBV) infection are essential for the clinical management of individuals infected with EBV. A calibrated quantitative real-time PCR assay for the measurement of EBV DNA of both EBV-1 and 2 subtypes was developed, combining the detection of the EBV DNA and a synthetic DNA calibrator in a multiplex PCR format. The assay displays a wide dynamic range and a high degree of accuracy even in the presence of 1μg of human genomic DNA. This assay measures with the same efficiency EBV DNA from strains prevalent in different geographic areas. The clinical sensitivity and specificity of the system were evaluated by testing 181 peripheral blood mononuclear cell (PBMCs) and plasma specimens obtained from 21 patients subjected to bone marrow transplantation, 70 HIV-seropositive subjects and 23 healthy controls. Patients affected by EBV-associated post-transplant lymphoprolipherative disorders had the highest frequency of EBV detection and the highest viral load. Persons infected with HIV had higher levels of EBV DNA load in PBMCs and a higher frequency of EBV plasma viremia compared to healthy controls. In conclusion, this new assay provides a reliable high-throughput method for the quantitation of EBV DNA in clinical samples. Copyright © 2011 Elsevier B.V. All rights reserved.

Friction coefficient of skin in real-time.

Science.gov (United States)

Sivamani, Raja K; Goodman, Jack; Gitis, Norm V; Maibach, Howard I

2003-08-01

Friction studies are useful in quantitatively investigating the skin surface. Previous studies utilized different apparatuses and materials for these investigations but there was no real-time test parameter control or monitoring. Our studies incorporated the commercially available UMT Series Micro-Tribometer, a tribology instrument that permits real-time monitoring and calculation of the important parameters in friction studies, increasing the accuracy over previous tribology and friction measurement devices used on skin. Our friction tests were performed on four healthy volunteers and on abdominal skin samples. A stainless steel ball was pressed on to the skin with at a pre-set load and then moved across the skin at a constant velocity of 5 mm/min. The UMT continuously monitored the friction force of the skin and the normal force of the ball to calculate the friction coefficient in real-time. Tests investigated the applicability of Amonton's law, the impact of increased and decreased hydration, and the effect of the application of moisturizers. The friction coefficient depends on the normal load applied, and Amonton's law does not provide an accurate description for the skin surface. Application of water to the skin increased the friction coefficient and application of isopropyl alcohol decreased it. Fast acting moisturizers immediately increased the friction coefficient, but did not have the prolonged effect of the slow, long lasting moisturizers. The UMT is capable of making real-time measurements on the skin and can be used as an effective tool to study friction properties. Results from the UMT measurements agree closely with theory regarding the skin surface.

Ruggedized downhole tool for real-time measurements and uses thereof

Energy Technology Data Exchange (ETDEWEB)

Hess, Ryan Falcone; Lindblom, Scott C.; Yelton, William G.; Limmer, Steven J.; Boyle, Timothy J.; Cieslewski, Grzegorz

2018-01-09

The present invention relates to ruggedized downhole tools and sensors, as well as uses thereof. In particular, these tools can operate under extreme conditions and, therefore, allow for real-time measurements in geothermal reservoirs or other potentially harsh environments. One exemplary sensor includes a ruggedized ion selective electrode (ISE) for detecting tracer concentrations in real-time. In one embodiment, the ISE includes a solid, non-conductive potting material and an ion selective material, which are disposed in a temperature-resistant electrode body. Other electrode configurations, tools, and methods are also described.

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

Development of a non invasion real-time PCR assay for the quantitation of chicken parvovirus in fecal swabs

Science.gov (United States)

The present study describes the development of a real time Taqman polymerase chain reaction (PCR) assay using a fluorescent labeled probe for the detection and quantitation of chicken parvovirus (ChPV) in feces. The primers and probes were designed based on the nucleotide sequence of the non struct...

MTF analysis of the near-real time neutron radiography facility at MURR

International Nuclear Information System (INIS)

Lindsay, J.T.; Alger, D.M.; Bull, S.R.; Miller, W.H.

1983-01-01

Several neutron radiography systems designed to view transient processes on a real-time basis have been developed. With the advent of these different real-time systems comes the necessity to develop a means to quantitatively evaluate and compare these systems. A suitable method for measuring the resolution capabilities of the image-forming system is the determination of the modulation transfer function (MTF). The MTF is a measure of an imaging system's ability to reproduce the spatial frequencies present in an image. The system in use at the University of Missouri Research Reactor is described. (Auth.)

Real-Time Inhibitor Recession Measurements in the Space Shuttle Reusable Solid Rocket Motors

Science.gov (United States)

McWhorter, Bruce B.; Ewing, Mark E.; McCool, Alex (Technical Monitor)

2001-01-01

Real-time char line recession measurements were made on propellant inhibitors of the Space Shuttle Reusable Solid Rocket Motor (RSRM). The RSRM FSM-8 static test motor propellant inhibitors (composed of a rubber insulation material) were successfully instrumented with eroding potentiometers and thermocouples. The data was used to establish inhibitor recession versus time relationships. Normally, pre-fire and post-fire insulation thickness measurements establish the thermal performance of an ablating insulation material. However, post-fire inhibitor decomposition and recession measurements are complicated by the fact that most of the inhibitor is back during motor operation. It is therefore a difficult task to evaluate the thermal protection offered by the inhibitor material. Real-time measurements would help this task. The instrumentation program for this static test motor marks the first time that real-time inhibitors. This report presents that data for the center and aft field joint forward facing inhibitors. The data was primarily used to measure char line recession of the forward face of the inhibitors which provides inhibitor thickness reduction versus time data. The data was also used to estimate the inhibitor height versus time relationship during motor operation.

Cloud computing platform for real-time measurement and verification of energy performance

International Nuclear Information System (INIS)

Ke, Ming-Tsun; Yeh, Chia-Hung; Su, Cheng-Jie

2017-01-01

Highlights: • Application of PSO algorithm can improve the accuracy of the baseline model. • M&V cloud platform automatically calculates energy performance. • M&V cloud platform can be applied in all energy conservation measures. • Real-time operational performance can be monitored through the proposed platform. • M&V cloud platform facilitates the development of EE programs and ESCO industries. - Abstract: Nations worldwide are vigorously promoting policies to improve energy efficiency. The use of measurement and verification (M&V) procedures to quantify energy performance is an essential topic in this field. Currently, energy performance M&V is accomplished via a combination of short-term on-site measurements and engineering calculations. This requires extensive amounts of time and labor and can result in a discrepancy between actual energy savings and calculated results. In addition, the M&V period typically lasts for periods as long as several months or up to a year, the failure to immediately detect abnormal energy performance not only decreases energy performance, results in the inability to make timely correction, and misses the best opportunity to adjust or repair equipment and systems. In this study, a cloud computing platform for the real-time M&V of energy performance is developed. On this platform, particle swarm optimization and multivariate regression analysis are used to construct accurate baseline models. Instantaneous and automatic calculations of the energy performance and access to long-term, cumulative information about the energy performance are provided via a feature that allows direct uploads of the energy consumption data. Finally, the feasibility of this real-time M&V cloud platform is tested for a case study involving improvements to a cold storage system in a hypermarket. Cloud computing platform for real-time energy performance M&V is applicable to any industry and energy conservation measure. With the M&V cloud platform, real-time

TaqMan MGB probe fluorescence real-time quantitative PCR for rapid detection of Chinese Sacbrood virus.

Directory of Open Access Journals (Sweden)

Ma Mingxiao

Full Text Available Sacbrood virus (SBV is a picorna-like virus that affects honey bees (Apis mellifera and results in the death of the larvae. Several procedures are available to detect Chinese SBV (CSBV in clinical samples, but not to estimate the level of CSBV infection. The aim of this study was develop an assay for rapid detection and quantification of this virus. Primers and probes were designed that were specific for CSBV structural protein genes. A TaqMan minor groove binder (MGB probe-based, fluorescence real-time quantitative PCR was established. The specificity, sensitivity and stability of the assay were assessed; specificity was high and there were no cross-reactivity with healthy larvae or other bee viruses. The assay was applied to detect CSBV in 37 clinical samples and its efficiency was compared with clinical diagnosis, electron microscopy observation, and conventional RT-PCR. The TaqMan MGB-based probe fluorescence real-time quantitative PCR for CSBV was more sensitive than other methods tested. This assay was a reliable, fast, and sensitive method that was used successfully to detect CSBV in clinical samples. The technology can provide a useful tool for rapid detection of CSBV. This study has established a useful protocol for CSBV testing, epidemiological investigation, and development of animal models.

Analytical real-time measurement of a three-dimensional weld pool surface

International Nuclear Information System (INIS)

Zhang, WeiJie; Zhang, YuMing; Wang, XueWu

2013-01-01

The ability to observe and measure weld pool surfaces in real-time is the core of the foundation for next generation intelligent welding that can partially imitate skilled welders who observe the weld pool to acquire information on the welding process. This study aims at the real-time measurement of the specular three-dimensional (3D) weld pool surface under a strong arc in gas tungsten arc welding (GTAW). An innovative vision system is utilized in this study to project a dot-matrix laser pattern on the specular weld pool surface. Its reflection from the surface is intercepted at a distance from the arc by a diffuse plane. The intercepted laser dots illuminate this plane producing an image showing the reflection pattern. The deformation of this reflection pattern from the projected pattern (e.g. the dot matrix) is used to derive the 3D shape of the reflection surface, i.e., the weld pool surface. Based on careful analysis, the underlying reconstruction problem is formulated mathematically. An analytic solution is proposed to solve this formulated problem resulting in the weld pool surface being reconstructed on average in 3.04 ms during welding experiments. A vision-based monitoring system is thus established to measure the weld pool surface in GTAW in real-time. In order to verify the effectiveness of the proposed reconstruction algorithm, first numerical simulation is conducted. The proposed algorithm is then tested on a spherical convex mirror with a priori knowledge of its geometry. The detailed analysis of the measurement error validates the accuracy of the proposed algorithm. Results from the real-time experiments verify the robustness of the proposed reconstruction algorithm. (paper)

Real-time and high accuracy frequency measurements for intermediate frequency narrowband signals

Science.gov (United States)

Tian, Jing; Meng, Xiaofeng; Nie, Jing; Lin, Liwei

2018-01-01

Real-time and accurate measurements of intermediate frequency signals based on microprocessors are difficult due to the computational complexity and limited time constraints. In this paper, a fast and precise methodology based on the sigma-delta modulator is designed and implemented by first generating the twiddle factors using the designed recursive scheme. This scheme requires zero times of multiplications and only half amounts of addition operations by using the discrete Fourier transform (DFT) and the combination of the Rife algorithm and Fourier coefficient interpolation as compared with conventional methods such as DFT and Fast Fourier Transform. Experimentally, when the sampling frequency is 10 MHz, the real-time frequency measurements with intermediate frequency and narrowband signals have a measurement mean squared error of ±2.4 Hz. Furthermore, a single measurement of the whole system only requires approximately 0.3 s to achieve fast iteration, high precision, and less calculation time.

Real-time long term measurement using integrated framework for ubiquitous smart monitoring

Science.gov (United States)

Heo, Gwanghee; Lee, Giu; Lee, Woosang; Jeon, Joonryong; Kim, Pil-Joong

2007-04-01

Ubiquitous monitoring combining internet technologies and wireless communication is one of the most promising technologies of infrastructure health monitoring against the natural of man-made hazards. In this paper, an integrated framework of the ubiquitous monitoring is developed for real-time long term measurement in internet environment. This framework develops a wireless sensor system based on Bluetooth technology and sends measured acceleration data to the host computer through TCP/IP protocol. And it is also designed to respond to the request of web user on real time basis. In order to verify this system, real time monitoring tests are carried out on a prototype self-anchored suspension bridge. Also, wireless measurement system is analyzed to estimate its sensing capacity and evaluate its performance for monitoring purpose. Based on the evaluation, this paper proposes the effective strategies for integrated framework in order to detect structural deficiencies and to design an early warning system.

Exploring Valid Reference Genes for Quantitative Real-Time PCR Analysis in Sesamia inferens (Lepidoptera: Noctuidae)

OpenAIRE

Sun, Meng; Lu, Ming-Xing; Tang, Xiao-Tian; Du, Yu-Zhou

2015-01-01

The pink stem borer, Sesamia inferens, which is endemic in China and other parts of Asia, is a major pest of rice and causes significant yield loss in this host plant. Very few studies have addressed gene expression in S. inferens. Quantitative real-time PCR (qRT-PCR) is currently the most accurate and sensitive method for gene expression analysis. In qRT-PCR, data are normalized using reference genes, which help control for internal differences and reduce error between samples. In this study...

[Quantitative fluorogenic real-time PCR assay for respiratory syncytial virus detection].

Science.gov (United States)

Zhang, Qi-wei; You, Shang-you; Sun, Ji-min; Wu, Qi; Yu, Chun-hua; Zhang, Chu-yu

2005-07-01

To Establish a rapid and objective quantitative fluorogenic real-time PCR assay for early detection of human respiratory syncytial virus (hRSV). Two pairs of primers and one TaqMan Fluorogenic probe that are specific for the recognition of the most conservative N gene of hRSV for virus detection with LighCycler PCR in 93 nasopharyngeal secretion specimens collected from infants and young children. The assay was compared with virus isolation, routine PCR, nested PCR, and enzyme-linked immunosorbent assay (ELISA). This TaqMan assay had a sensitivity of 1 x 10(2) cDNA copies/microl with a dynamic range between 1 x 10(2) and 1 x 10(7) cDNA copies/microl, which was the same as that of nested PCR, but 10 times more sensitive than routine PCR. The specificity of the assay was evaluated by comparing hRSV with polivirus type 1, coxsackie virus type 2, influenza A, influenza B and adenovirus type 7. A PCR product of the expected size (195 bp) was produced and fluorescence signal detected for hRSV, but not for any of the other viruses. The results in LightCycler and Rotor-Gene instrument were consistent. Forty-four specimens (43.9%) were hRSV-positive with this assay and 4 (4/93,4.3%) were hRSV-positive with ELISA, showing rather low correlation between the two methods. No visible relation was found between the concentration of hRSV RNA and severity of the disease. This assay is rapid, sensitive, specific and quantitative, and has the potential of wide application for early diagnosis of hRSV infection and evaluation of the therapeutic effect.

A real-time, quantitative PCR protocol for assessing the relative parasitemia of Leucocytozoon in waterfowl

Science.gov (United States)

Smith, Matthew M.; Schmutz, Joel A.; Apelgren, Chloe; Ramey, Andy M.

2015-01-01

Microscopic examination of blood smears can be effective at diagnosing and quantifying hematozoa infections. However, this method requires highly trained observers, is time consuming, and may be inaccurate for detection of infections at low levels of parasitemia. To develop a molecular methodology for identifying and quantifying Leucocytozoon parasite infection in wild waterfowl (Anseriformes), we designed a real-time, quantitative PCR protocol to amplify Leucocytozoon mitochondrial DNA using TaqMan fluorogenic probes and validated our methodology using blood samples collected from waterfowl in interior Alaska during late summer and autumn (n = 105). By comparing our qPCR results to those derived from a widely used nested PCR protocol, we determined that our assay showed high levels of sensitivity (91%) and specificity (100%) in detecting Leucocytozoon DNA from host blood samples. Additionally, results of a linear regression revealed significant correlation between the raw measure of parasitemia produced by our qPCR assay (Ct values) and numbers of parasites observed on blood smears (R2 = 0.694, P = 0.003), indicating that our assay can reliably determine the relative parasitemia levels among samples. This methodology provides a powerful new tool for studies assessing effects of haemosporidian infection in wild avian species.

Real-time scatter measurement and correction in film radiography

International Nuclear Information System (INIS)

Shaw, C.G.

1987-01-01

A technique for real-time scatter measurement and correction in scanning film radiography is described. With this technique, collimated x-ray fan beams are used to partially reject scattered radiation. Photodiodes are attached to the aft-collimator for sampled scatter measurement. Such measurement allows the scatter distribution to be reconstructed and subtracted from digitized film image data for accurate transmission measurement. In this presentation the authors discuss the physical and technical considerations of this scatter correction technique. Examples are shown that demonstrate the feasibility of the technique. Improved x-ray transmission measurement and dual-energy subtraction imaging are demonstrated with phantoms

AUTOMATED CONTROL AND REAL-TIME DATA PROCESSING OF WIRE SCANNER/HALO SCRAPER MEASUREMENTS

International Nuclear Information System (INIS)

Day, L.A.; Gilpatrick, J.D.

2001-01-01

The Low-Energy Demonstration Accelerator (LEDA), assembled and operating at Los Alamos National Laboratory, provides the platform for obtaining measurements of high-power proton beam-halo formation. Control system software and hardware have been integrated and customized to enable the production of real-time beam-halo profiles. The Experimental Physics and Industrial Control System (EPICS) hosted on a VXI platform, Interactive Data Language (IDL) programs hosted on UNIX platforms, and LabVIEW (LV) Virtual Instruments hosted on a PC platform have been integrated and customized to provide real-time, synchronous motor control, data acquisition, and data analysis of data acquired through specialized DSP instrumentation. These modules communicate through EPICS Channel Access (CA) communication protocol extensions to control and manage execution flow ensuring synchronous data acquisition and real-time processing of measurement data. This paper describes the software integration and management scheme implemented to produce these real-time beam profiles

MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments

NARCIS (Netherlands)

Bustin, S.A.; Beaulieu, J.F.; Huggett, J.; Jaggi, R.; Kibenge, F.S.; Olsvik, P.A.; Penning, L.C.; Toegel, S.

2010-01-01

MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments Stephen A Bustin1 , Jean-François Beaulieu2 , Jim Huggett3 , Rolf Jaggi4 , Frederick SB Kibenge5 , Pål A Olsvik6 , Louis C Penning7 and Stefan Toegel8 1 Centre for

Real-Time Mapping Spectroscopy on the Ground, in the Air, and in Space

Science.gov (United States)

Thompson, D. R.; Allwood, A.; Chien, S.; Green, R. O.; Wettergreen, D. S.

2016-12-01

Real-time data interpretation can benefit both remote in situ exploration and remote sensing. Basic analyses at the sensor can monitor instrument performance and reveal invisible science phenomena in real time. This promotes situational awareness for remote robotic explorers or campaign decision makers, enabling adaptive data collection, reduced downlink requirements, and coordinated multi-instrument observations. Fast analysis is ideal for mapping spectrometers providing unambiguous, quantitative geophysical measurements. This presentation surveys recent computational advances in real-time spectroscopic analysis for Earth science and planetary exploration. Spectral analysis at the sensor enables new operations concepts that significantly improve science yield. Applications include real-time detection of fugitive greenhouse emissions by airborne monitoring, real-time cloud screening and mineralogical mapping by orbital spectrometers, and adaptive measurement by the PIXL instrument on the Mars 2020 rover. Copyright 2016 California Institute of Technology. All Rights Reserved. We acknowledge support of the US Government, NASA, the Earth Science Division and Terrestrial Ecology program.

Real-time diamagnetic flux measurements on ASDEX Upgrade.

Science.gov (United States)

Giannone, L; Geiger, B; Bilato, R; Maraschek, M; Odstrčil, T; Fischer, R; Fuchs, J C; McCarthy, P J; Mertens, V; Schuhbeck, K H

2016-05-01

Real-time diamagnetic flux measurements are now available on ASDEX Upgrade. In contrast to the majority of diamagnetic flux measurements on other tokamaks, no analog summation of signals is necessary for measuring the change in toroidal flux or for removing contributions arising from unwanted coupling to the plasma and poloidal field coil currents. To achieve the highest possible sensitivity, the diamagnetic measurement and compensation coil integrators are triggered shortly before plasma initiation when the toroidal field coil current is close to its maximum. In this way, the integration time can be chosen to measure only the small changes in flux due to the presence of plasma. Two identical plasma discharges with positive and negative magnetic field have shown that the alignment error with respect to the plasma current is negligible. The measured diamagnetic flux is compared to that predicted by TRANSP simulations. The poloidal beta inferred from the diamagnetic flux measurement is compared to the values calculated from magnetic equilibrium reconstruction codes. The diamagnetic flux measurement and TRANSP simulation can be used together to estimate the coupled power in discharges with dominant ion cyclotron resonance heating.

Chemisorption of iodine-125 to gold nanoparticles allows for real-time quantitation and potential use in nanomedicine

Energy Technology Data Exchange (ETDEWEB)

Walsh, Adrian A, E-mail: a.walsh@nanobiosols.com [Liverpool Science Park, Nano Biosols Ltd (United Kingdom)

2017-04-15

Gold nanoparticles have been available for many years as a research tool in the life sciences due to their electron density and optical properties. New applications are continually being developed, particularly in nanomedicine. One drawback is the need for an easy, real-time quantitation method for gold nanoparticles so that the effects observed in in vitro cell toxicity assays and cell uptake studies can be interpreted quantitatively in terms of nanoparticle loading. One potential method of quantifying gold nanoparticles in real time is by chemisorption of iodine-125, a gamma emitter, to the nanoparticles. This paper revisits the labelling of gold nanoparticles with iodine-125, first described 30 years ago and never fully exploited since. We explore the chemical properties and usefulness in quantifying bio-functionalised gold nanoparticle binding in a quick and simple manner. The gold particles were labelled specifically and quantitatively simply by mixing the two items. The nature of the labelling is chemisorption and is robust, remaining bound over several weeks in a variety of cell culture media. Chemisorption was confirmed as potassium iodide can remove the label whereas sodium chloride and many other buffers had no effect. Particles precoated in polymers or proteins can be labelled just as efficiently allowing for post-labelling experiments in situ rather than using radioactive gold atoms in the production process. We also demonstrate that interparticle exchange of I-125 between different size particles does not appear to take place confirming the affinity of the binding.

Chemisorption of iodine-125 to gold nanoparticles allows for real-time quantitation and potential use in nanomedicine

Science.gov (United States)

Walsh, Adrian A.

2017-04-01

Gold nanoparticles have been available for many years as a research tool in the life sciences due to their electron density and optical properties. New applications are continually being developed, particularly in nanomedicine. One drawback is the need for an easy, real-time quantitation method for gold nanoparticles so that the effects observed in in vitro cell toxicity assays and cell uptake studies can be interpreted quantitatively in terms of nanoparticle loading. One potential method of quantifying gold nanoparticles in real time is by chemisorption of iodine-125, a gamma emitter, to the nanoparticles. This paper revisits the labelling of gold nanoparticles with iodine-125, first described 30 years ago and never fully exploited since. We explore the chemical properties and usefulness in quantifying bio-functionalised gold nanoparticle binding in a quick and simple manner. The gold particles were labelled specifically and quantitatively simply by mixing the two items. The nature of the labelling is chemisorption and is robust, remaining bound over several weeks in a variety of cell culture media. Chemisorption was confirmed as potassium iodide can remove the label whereas sodium chloride and many other buffers had no effect. Particles precoated in polymers or proteins can be labelled just as efficiently allowing for post-labelling experiments in situ rather than using radioactive gold atoms in the production process. We also demonstrate that interparticle exchange of I-125 between different size particles does not appear to take place confirming the affinity of the binding.

Chemisorption of iodine-125 to gold nanoparticles allows for real-time quantitation and potential use in nanomedicine

International Nuclear Information System (INIS)

Walsh, Adrian A

2017-01-01

Gold nanoparticles have been available for many years as a research tool in the life sciences due to their electron density and optical properties. New applications are continually being developed, particularly in nanomedicine. One drawback is the need for an easy, real-time quantitation method for gold nanoparticles so that the effects observed in in vitro cell toxicity assays and cell uptake studies can be interpreted quantitatively in terms of nanoparticle loading. One potential method of quantifying gold nanoparticles in real time is by chemisorption of iodine-125, a gamma emitter, to the nanoparticles. This paper revisits the labelling of gold nanoparticles with iodine-125, first described 30 years ago and never fully exploited since. We explore the chemical properties and usefulness in quantifying bio-functionalised gold nanoparticle binding in a quick and simple manner. The gold particles were labelled specifically and quantitatively simply by mixing the two items. The nature of the labelling is chemisorption and is robust, remaining bound over several weeks in a variety of cell culture media. Chemisorption was confirmed as potassium iodide can remove the label whereas sodium chloride and many other buffers had no effect. Particles precoated in polymers or proteins can be labelled just as efficiently allowing for post-labelling experiments in situ rather than using radioactive gold atoms in the production process. We also demonstrate that interparticle exchange of I-125 between different size particles does not appear to take place confirming the affinity of the binding.

Real-time, in vivo measurement of radiation dose during radioimmunotherapy in mice using a miniature MOSFET dosimeter probe

International Nuclear Information System (INIS)

Gladstone, D.J.; Chin, L.M.

1995-01-01

This report presents the first real-time measurement of absorbed radiation dose during radioimmunotherapy in mice. Dose rate and total dose at the center of the tumor were measured after administration of 90 Y-labeled antibodies using a miniature metal oxide semiconductor field-effect transistor radiation dosimeter probe which was inserted into the center of the tumor volume. Continuous real-time measurements were made for as long as 23 h after injection of the radiolabeled antibodies. Comparison of the real-time dose-rate measurements with estimates based on the MIRD formalism indicates good agreement. The real-time measurements are further compared to measurements made in a second experiment in which groups of mice were sacrificed at individual times after injection of the same radiolabeled antibodies. The real-time measurements agree well with the measurements in excised tumors. The real-time measurements have greater time resolution and are much more efficient than traditional uptake measurements. 17 refs., 2 figs

Coexistence of Epstein-Barr virus and Parvovirus B19 in tonsillar tissue samples: quantitative measurement by real-time PCR.

Science.gov (United States)

Sahiner, Fatih; Gümral, Ramazan; Yildizoğlu, Üzeyir; Babayiğit, Mustafa Alparslan; Durmaz, Abdullah; Yiğit, Nuri; Saraçli, Mehmet Ali; Kubar, Ayhan

2014-08-01

In this study, we aimed to investigate the presence and copy number of six different viruses in tonsillar tissue samples removed surgically because of chronic recurrent tonsillitis or chronic obstructive tonsillar hypertrophy. In total, 56 tissue samples (tonsillar core) collected from 44 children and 12 adults were included in this study. The presence of viruses was investigated using a new TaqMan-based quantitative real-time PCR assay. Of the 56 tissue samples, 67.9% (38/56) were positive for at least one of the six viruses. Epstein-Barr virus was the most frequently detected virus, being found in 53.6% (30/56), followed by human Parvovirus B19 21.4% (12/56), human adenovirus 12.5% (7/56), human Cytomegalovirus 5.4% (3/56), BK polyomavirus 1.8% (1/56), and Herpes simplex virus 1.8% (1/56). Precancerous or cancerous changes were not detected in the tonsillar tissue samples by pathologic examination, whereas lymphoid hyperplasia was observed in 24 patients. In contrast to other viruses, B19 virus was present in high copy number in tonsillar tissues. The rates of EBV and B19 virus with high copy number (>500.000 copies/ml) were higher in children than in adults, and a positive relationship was also found between the presence of EBV and the presence of B19 virus with high copy number (P=0.037). It is previously reported that some viral agents are associated with different chronic tonsillar pathologies. In the present study, the presence of B19 virus in tonsillar core samples was investigated quantitatively for the first time, and our data suggests that EBV infections could be associated with B19 virus infections or could facilitate B19 virus replication. However, further detailed studies are needed to clarify this observation. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Quantitation of RHD by real-time polymerase chain reaction for determination of RHD zygosity and RHD mosaicism/chimerism

DEFF Research Database (Denmark)

Krog, Grethe Risum; Clausen, Frederik Banch; Dziegiel, Morten Hanefeld

2007-01-01

Determination of RHD zygosity of the spouse is crucial in preconception counseling of families with history of hemolytic disease of the fetus and newborn caused by anti-D. RHD zygosity can be determined by quantitative real-time polymerase chain reaction (PCR) basically by determining RHD dosage,......, and this feature is relevant in investigating RHD mosaicism and chimerism....

Measurement of the real time fill-pattern at the Australian Synchrotron

International Nuclear Information System (INIS)

Peake, D.J.; Boland, M.J.; LeBlanc, G.S.; Rassool, R.P.

2008-01-01

This article describes the development, commissioning and operation of a Fill-Pattern Monitor (FPM) for the Australian Synchrotron that measures the real-time intensity distribution of the electron bunches in the storage ring. Using a combination of an ultra-fast photodiode and a high-speed digitiser, real-time measurement of the fill-pattern at bunch-by-bunch resolution was achieved. The results compare very well with current methods of measuring the fill-pattern, such as a pick-up style detector. In addition, the FPM is fully integrated into the EPICS control system. The data provided by the FPM gives accurate RF bucket position and relative bunch currents over a wide range of stored beam currents, from 0.01 mA in a single bunch to 200 mA total beam current. The FPM monitors the success of an injection attempt into the storage ring and is used in a feedback loop to determine where to target the next injection. Using the FPM a beam top-up mode was successfully tested, resulting in a near constant beam current by periodic targeted injections over an 8 h shift. Results are presented for dynamically topped up real-time injection, where the beam pattern was squared using an intensity-dependent injection algorithm

Real-time micro-vibration multi-spot synchronous measurement within a region based on heterodyne interference

Science.gov (United States)

Lan, Ma; Xiao, Wen; Chen, Zonghui; Hao, Hongliang; Pan, Feng

2018-01-01

Real-time micro-vibration measurement is widely used in engineering applications. It is very difficult for traditional optical detection methods to achieve real-time need in a relatively high frequency and multi-spot synchronous measurement of a region at the same time,especially at the nanoscale. Based on the method of heterodyne interference, an experimental system of real-time measurement of micro - vibration is constructed to satisfy the demand in engineering applications. The vibration response signal is measured by combing optical heterodyne interferometry and a high-speed CMOS-DVR image acquisition system. Then, by extracting and processing multiple pixels at the same time, four digital demodulation technique are implemented to simultaneously acquire the vibrating velocity of the target from the recorded sequences of images. Different kinds of demodulation algorithms are analyzed and the results show that these four demodulation algorithms are suitable for different interference signals. Both autocorrelation algorithm and cross-correlation algorithm meet the needs of real-time measurements. The autocorrelation algorithm demodulates the frequency more accurately, while the cross-correlation algorithm is more accurate in solving the amplitude.

Real-time PCR assays for the quantitation of rDNA from apricot and other plant species in marzipan.

Science.gov (United States)

Haase, Ilka; Brüning, Philipp; Matissek, Reinhard; Fischer, Markus

2013-04-10

Marzipan or marzipan raw paste is a typical German sweet which is consumed directly or is used as an ingredient in the bakery industry/confectionery (e.g., in stollen) and as filling for chocolate candies. Almonds (blanched and pealed) and sugar are the only ingredients for marzipan production according to German food guidelines. Especially for the confectionery industry, the use of persipan, which contains apricot or peach kernels instead of almonds, is preferred due to its stronger aroma. In most of the companies, both raw pastes are produced, in most cases on the same production line, running the risk of an unintended cross contamination. Additionally, due to high almond market values, dilutions of marzipan with cheaper seeds may occur. Especially in the case of apricot and almond, the close relationship of both species is a challenge for the analysis. DNA based methods for the qualitative detection of apricot, peach, pea, bean, lupine, soy, cashew, pistachio, and chickpea in marzipan have recently been published. In this study, different quantitation strategies on the basis of real-time PCR have been evaluated and a relative quantitation method with a reference amplification product was shown to give the best results. As the real-time PCR is based on the high copy rDNA-cluster, even contaminations <1% can be reliably quantitated.

Real-Time Alpine Measurement System Using Wireless Sensor Networks

Directory of Open Access Journals (Sweden)

Sami A. Malek

2017-11-01

Full Text Available Monitoring the snow pack is crucial for many stakeholders, whether for hydro-power optimization, water management or flood control. Traditional forecasting relies on regression methods, which often results in snow melt runoff predictions of low accuracy in non-average years. Existing ground-based real-time measurement systems do not cover enough physiographic variability and are mostly installed at low elevations. We present the hardware and software design of a state-of-the-art distributed Wireless Sensor Network (WSN-based autonomous measurement system with real-time remote data transmission that gathers data of snow depth, air temperature, air relative humidity, soil moisture, soil temperature, and solar radiation in physiographically representative locations. Elevation, aspect, slope and vegetation are used to select network locations, and distribute sensors throughout a given network location, since they govern snow pack variability at various scales. Three WSNs were installed in the Sierra Nevada of Northern California throughout the North Fork of the Feather River, upstream of the Oroville dam and multiple powerhouses along the river. The WSNs gathered hydrologic variables and network health statistics throughout the 2017 water year, one of northern Sierra’s wettest years on record. These networks leverage an ultra-low-power wireless technology to interconnect their components and offer recovery features, resilience to data loss due to weather and wildlife disturbances and real-time topological visualizations of the network health. Data show considerable spatial variability of snow depth, even within a 1 km 2 network location. Combined with existing systems, these WSNs can better detect precipitation timing and phase in, monitor sub-daily dynamics of infiltration and surface runoff during precipitation or snow melt, and inform hydro power managers about actual ablation and end-of-season date across the landscape.

Real-Time Alpine Measurement System Using Wireless Sensor Networks.

Science.gov (United States)

Malek, Sami A; Avanzi, Francesco; Brun-Laguna, Keoma; Maurer, Tessa; Oroza, Carlos A; Hartsough, Peter C; Watteyne, Thomas; Glaser, Steven D

2017-11-09

Monitoring the snow pack is crucial for many stakeholders, whether for hydro-power optimization, water management or flood control. Traditional forecasting relies on regression methods, which often results in snow melt runoff predictions of low accuracy in non-average years. Existing ground-based real-time measurement systems do not cover enough physiographic variability and are mostly installed at low elevations. We present the hardware and software design of a state-of-the-art distributed Wireless Sensor Network (WSN)-based autonomous measurement system with real-time remote data transmission that gathers data of snow depth, air temperature, air relative humidity, soil moisture, soil temperature, and solar radiation in physiographically representative locations. Elevation, aspect, slope and vegetation are used to select network locations, and distribute sensors throughout a given network location, since they govern snow pack variability at various scales. Three WSNs were installed in the Sierra Nevada of Northern California throughout the North Fork of the Feather River, upstream of the Oroville dam and multiple powerhouses along the river. The WSNs gathered hydrologic variables and network health statistics throughout the 2017 water year, one of northern Sierra's wettest years on record. These networks leverage an ultra-low-power wireless technology to interconnect their components and offer recovery features, resilience to data loss due to weather and wildlife disturbances and real-time topological visualizations of the network health. Data show considerable spatial variability of snow depth, even within a 1 km 2 network location. Combined with existing systems, these WSNs can better detect precipitation timing and phase in, monitor sub-daily dynamics of infiltration and surface runoff during precipitation or snow melt, and inform hydro power managers about actual ablation and end-of-season date across the landscape.

Real-time and quantitative isotropic spatial resolution susceptibility imaging for magnetic nanoparticles

Science.gov (United States)

Pi, Shiqiang; Liu, Wenzhong; Jiang, Tao

2018-03-01

The magnetic transparency of biological tissue allows the magnetic nanoparticle (MNP) to be a promising functional sensor and contrast agent. The complex susceptibility of MNPs, strongly influenced by particle concentration, excitation magnetic field and their surrounding microenvironment, provides significant implications for biomedical applications. Therefore, magnetic susceptibility imaging of high spatial resolution will give more detailed information during the process of MNP-aided diagnosis and therapy. In this study, we present a novel spatial magnetic susceptibility extraction method for MNPs under a gradient magnetic field, a low-frequency drive magnetic field, and a weak strength high-frequency magnetic field. Based on this novel method, a magnetic particle susceptibility imaging (MPSI) of millimeter-level spatial resolution (<3 mm) was achieved using our homemade imaging system. Corroborated by the experimental results, the MPSI shows real-time (1 s per frame acquisition) and quantitative abilities, and isotropic high resolution.

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

Directory of Open Access Journals (Sweden)

Dial Stacey L

2008-07-01

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

Quantitative EEG analysis using error reduction ratio-causality test; validation on simulated and real EEG data.

Science.gov (United States)

Sarrigiannis, Ptolemaios G; Zhao, Yifan; Wei, Hua-Liang; Billings, Stephen A; Fotheringham, Jayne; Hadjivassiliou, Marios

2014-01-01

To introduce a new method of quantitative EEG analysis in the time domain, the error reduction ratio (ERR)-causality test. To compare performance against cross-correlation and coherence with phase measures. A simulation example was used as a gold standard to assess the performance of ERR-causality, against cross-correlation and coherence. The methods were then applied to real EEG data. Analysis of both simulated and real EEG data demonstrates that ERR-causality successfully detects dynamically evolving changes between two signals, with very high time resolution, dependent on the sampling rate of the data. Our method can properly detect both linear and non-linear effects, encountered during analysis of focal and generalised seizures. We introduce a new quantitative EEG method of analysis. It detects real time levels of synchronisation in the linear and non-linear domains. It computes directionality of information flow with corresponding time lags. This novel dynamic real time EEG signal analysis unveils hidden neural network interactions with a very high time resolution. These interactions cannot be adequately resolved by the traditional methods of coherence and cross-correlation, which provide limited results in the presence of non-linear effects and lack fidelity for changes appearing over small periods of time. Copyright © 2013 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

Dynamic fiber Bragg grating strain sensor interrogation with real-time measurement

Science.gov (United States)

Park, Jinwoo; Kwon, Yong Seok; Ko, Myeong Ock; Jeon, Min Yong

2017-11-01

We demonstrate a 1550 nm band resonance Fourier-domain mode-locked (FDML) fiber laser with fiber Bragg grating (FBG) array. Using the FDML fiber laser, we successfully demonstrate real-time monitoring of dynamic FBG strain sensor interrogation for structural health monitoring. The resonance FDML fiber laser consists of six multiplexed FBGs, which are arranged in series with delay fiber lengths. It is operated by driving the fiber Fabry-Perot tunable filter (FFP-TF) with a sinusoidal waveform at a frequency corresponding to the round-trip time of the laser cavity. Each FBG forms a laser cavity independently in the FDML fiber laser because the light travels different length for each FBG. The very closely positioned two FBGs in a pair are operated simultaneously with a frequency in the FDML fiber laser. The spatial positions of the sensing pair can be distinguished from the variation of the applied frequency to the FFP-TF. One of the FBGs in the pair is used as a reference signal and the other one is fixed on the piezoelectric transducer stack to apply the dynamic strain. We successfully achieve real-time measurement of the abrupt change of the frequencies applied to the FBG without any signal processing delay. The real-time monitoring system is displayed simultaneously on the monitor for the variation of the two peaks, the modulation interval of the two peaks, and their fast Fourier transform spectrum. The frequency resolution of the dynamic variation could reach up to 0.5 Hz for 2 s integration time. It depends on the integration time to measure the dynamic variation. We believe that the real-time monitoring system will have a potential application for structural health monitoring.

Atomic force microscope cantilever as an encoding sensor for real-time displacement measurement

International Nuclear Information System (INIS)

Chen, Xiaomei; Koenders, Ludger; Wolff, Helmut; Haertig, Frank; Schilling, Meinhard

2010-01-01

A tuning fork-based atomic force microscope cantilever has been investigated for application as an encoding sensor for real-time displacement measurement. The algorithm used to encode the displacement is based on the direct count of the integer pitches of a known grating, and the calculation of the fractional parts of a pitch at the beginning and during displacement. A cross-correlation technique has been adopted and applied to the real-time signal filtering process for the determination of the pitch during scanning by using a half sinusoidal waveform template. For the first investigation, a 1D sinusoidal grating with the pitch of 300 nm is used. The repeatability of displacement measurements over a distance of 70 µm is better than 2.2 nm. As the first application, the real-time displacement of a scanning stage is measured by the new encoding principle as it is moved in an open-loop mode and closed-loop mode based on its built-in capacitance sensor

In-situ real time measurements of net erosion rates of copper during hydrogen plasma exposure

Science.gov (United States)

Kesler, Leigh; Wright, Graham; Peterson, Ethan; Whyte, Dennis

2013-10-01

In order to properly understand the dynamics of net erosion/deposition in fusion reactors, such as tokamaks, a diagnostic measuring the real time rates of net erosion/deposition during plasma exposure is necessary. The DIONISOS experiment produces real time measurements of net erosion/deposition by using Rutherford backscattering spectroscopy (RBS) ion beam analysis simultaneously with plasma exposure from a helicon plasma source. This in-situ method improves on ex-situ weight loss measurements by allowing measurement of possible synergistic effects of high ion implantation rates and net erosion rate and by giving a real time response to changes in plasma parameters. Previous work has validated this new technique for measuring copper (Cu) erosion from helium (He) plasma ion bombardment. This technique is now extended to measure copper erosion due to deuterium and hydrogen plasma ion exposure. Targets used were a 1.5 μm Cu layer on an aluminum substrate. Cu layer thickness is tracked in real time using 1.2 MeV proton RBS. Measured erosion rates will be compared to results from literature and He erosion rates. Supported by US DoE award DE-SC00-02060.

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.

Real-time particle volume fraction measurement in centrifuges by wireless electrical resistance detector

International Nuclear Information System (INIS)

Nagae, Fumiya; Okawa, Kazuya; Matsuno, Shinsuke; Takei, Masahiro; Zhao Tong; Ichijo, Noriaki

2015-01-01

In this study, wireless electrical resistance detector is developed as first step in order to develop electrical resistance tomography (ERT) that are attached wireless communication, and miniaturized. And the particle volume fraction measurement results appropriateness is qualitatively examined. The real-time particle volume fraction measurement is essential for centrifuges, because rotational velocity and supply should be controlled based on the results in order to obtain the effective separation, shorten process time and save energy. However, a technique for the particle volume fraction measurement in centrifuges has not existed yet. In other words, the real-time particle volume fraction measurement in centrifuges becomes innovative technologies. The experiment device reproduces centrifugation in two-phase using particle and salt solution as measuring object. The particle concentration is measured changing rotational velocity, supply and measurement section position. The measured concentration changes coincide with anticipated tendency of concentration changes. Therefore the particle volume fraction measurement results appropriateness are qualitatively indicated. (author)

Expression analysis of fusarium wilt resistance gene in melon by real-time quantitative pcr

International Nuclear Information System (INIS)

Wang, X.; Xu, B.; Zhao, L.; Gao, P.; Luan, F.

2014-01-01

Melon Actin gene was used as a reference gene, to explore the gene expression profiles of the Fom-2 gene in roots, stems, and leaves of melon MR-1 under induction by Fusarium oxysporum f. sp. melonis. Monitoring using real-time quantitative PCR showed similar accumulation patterns of Fom-2 in roots, stems, and leaves over the observation period of 1 to 11 days; the expression level in stems was the highest. The expression of the Fom-2 gene was strengthened by the prolongation of induction time. In stems, the expression of Fom-2 was 5.737 times higher than in the control at three days; in roots, expression of Fom-2 was 5.617 times higher than in the control at five days. Similarly, the expression of Fom-2 in leaves obviously increased. It was 4.441 times higher than in the control at 5 days. The expression of Fom-2 was non-tissue specific, up-regulated under induction by Fusarium, and related to early resistance to Fusarium wilt. (author)

Fluoro-luminometric real-time measurement of bacterial viability and killing.

Science.gov (United States)

Lehtinen, Janne; Virta, Marko; Lilius, Esa Matti

2003-10-01

The viability and killing of Escherichia coli was measured on a real-time basis using a fluoro-luminometric device, which allows successive measurements of fluorescence and bioluminescence without user intervention. Bacteria were made fluorescent and bioluminescent by expression of gfp and insect luciferase (lucFF) genes. The green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein, which accumulates in cells during growth, and therefore the measured fluorescence signal was proportional to the total number of cells. The luciferase reaction is dependent of ATP produced by living cells, so that the bioluminescence level was a direct measure of the viable cells. In contrast to the bacterial luciferase, the insect luciferase uses a water-soluble and nonvolatile substrate, which makes automated multi-well microplate assay possible. For the validation of the assay, the proportion of living and dead cell populations was experimentally modified by incubating E. coli cells in the presence of various ethanol concentrations. Bacterial viability and killing measured by a fluoro-luminometric assay correlated fairly well with the reference methods: conventional plate counting, optical density measurement and various flow cytometric analyses. The real-time assay described here allows following the changes in bacterial cultures and assessing the bactericidal and other effects of various chemical, immunological and physical agents simultaneously in large numbers of samples.

Verus: A Tool for Quantitative Analysis of Finite-State Real-Time Systems.

Science.gov (United States)

1996-08-12

Symbolic model checking is a technique for verifying finite-state concurrent systems that has been extended to handle real - time systems . Models with...up to 10(exp 30) states can often be verified in minutes. In this paper, we present a new tool to analyze real - time systems , based on this technique...We have designed a language, called Verus, for the description of real - time systems . Such a description is compiled into a state-transition graph and

Comparison of Abbott and Da-an real-time PCR for quantitating serum HBV DNA.

Science.gov (United States)

Qiu, Ning; Li, Rui; Yu, Jian-Guo; Yang, Wen; Zhang, Wei; An, Yong; Li, Tong; Liu, Xue-En; Zhuang, Hui

2014-09-07

To compare the performance of the Da-an real-time hepatitis B virus (HBV) DNA assay and Abbott RealTime HBV assay. HBV DNA standards as well as a total of 180 clinical serum samples from patients with chronic hepatitis B were measured using the Abbott and Da-an real-time polymerase chain reaction (PCR) assays. Correlation and Bland-Altman plot analysis was used to compare the performance of the Abbott and Da-an assays. The HBV DNA levels were logarithmically transformed for analysis. All statistical analyses were performed using SPSS for Windows version 18.0. The correlation between the two assays was analyzed by Pearson's correlation and linear regression. The Bland-Altman plots were used for the analysis of agreement between the two assays. A P value of Da-an assay were significantly correlated with the expected values of HBV DNA standards (r = 0.999, for Abbott; r = 0.987, for Da-an, P Da-an assay. Moreover, HBV DNA levels measured by the Abbott assay were significantly higher than those of the Da-an assay (6.23 ± 1.76 log IU/mL vs 5.46 ± 1.55 log IU/mL, P Da-an assay presented lower sensitivity and a narrower linear range as compared to the Abbott assay, suggesting the need to be improved.

Real-time microscopic 3D shape measurement based on optimized pulse-width-modulation binary fringe projection

Science.gov (United States)

Hu, Yan; Chen, Qian; Feng, Shijie; Tao, Tianyang; Li, Hui; Zuo, Chao

2017-07-01

In recent years, tremendous progress has been made in 3D measurement techniques, contributing to the realization of faster and more accurate 3D measurement. As a representative of these techniques, fringe projection profilometry (FPP) has become a commonly used method for real-time 3D measurement, such as real-time quality control and online inspection. To date, most related research has been concerned with macroscopic 3D measurement, but microscopic 3D measurement, especially real-time microscopic 3D measurement, is rarely reported. However, microscopic 3D measurement plays an important role in 3D metrology and is indispensable in some applications in measuring micro scale objects like the accurate metrology of MEMS components of the final devices to ensure their proper performance. In this paper, we proposed a method which effectively combines optimized binary structured patterns with a number-theoretical phase unwrapping algorithm to realize real-time microscopic 3D measurement. A slight defocusing of our optimized binary patterns can considerably alleviate the measurement error based on four-step phase-shifting FPP, providing the binary patterns with a comparable performance to ideal sinusoidal patterns. The static measurement accuracy can reach 8 μm, and the experimental results of a vibrating earphone diaphragm reveal that our system can successfully realize real-time 3D measurement of 120 frames per second (FPS) with a measurement range of 8~\\text{mm}× 6~\\text{mm} in lateral and 8 mm in depth.

A real-time BWR [boiling water reactor] stability measurement system

International Nuclear Information System (INIS)

March-Leuba, J.; King, W.T.

1987-01-01

This paper describes the characteristics of a portable, real-time system used for nonperturbational measurements of stability in boiling water reactors. The algorithm used in this system estimates the closed-loop asymptotic decay ratio using only the naturally occurring neutron noise and it is based on the univariate autoregressive methodology

Evaluation of Real-time Measurement Liver Tumor's Movement and SynchronyTM System's Accuracy of Radiosurgery using a Robot CyberKnife

International Nuclear Information System (INIS)

Kim, Gha Jung; Shim, Su Jung; Kim, Jeong Ho; Min, Chul Kee; Chung, Weon Kuu

2008-01-01

This study aimed to quantitatively measure the movement of tumors in real-time and evaluate the treatment accuracy, during the treatment of a liver tumor patient, who underwent radiosurgery with a Synchrony Respiratory motion tracking system of a robot CyberKnife. Materials and Methods: The study subjects included 24 liver tumor patients who underwent CyberKnife treatment, which included 64 times of treatment with the Synchrony Respiratory motion tracking system (SynchronyTM). The treatment involved inserting 4 to 6 acupuncture needles into the vicinity of the liver tumor in all the patients using ultrasonography as a guide. A treatment plan was set up using the CT images for treatment planning uses. The position of the acupuncture needle was identified for every treatment time by Digitally Reconstructed Radiography (DRR) prepared at the time of treatment planning and X-ray images photographed in real-time. Subsequent results were stored through a Motion Tracking System (MTS) using the Mtsmain.log treatment file. In this way, movement of the tumor was measured. Besides, the accuracy of radiosurgery using CyberKnife was evaluated by the correlation errors between the real-time positions of the acupuncture needles and the predicted coordinates. Results: The maximum and the average translational movement of the liver tumor were measured 23.5 mm and 13.9±5.5 mm, respectively from the superior to the inferior direction, 3.9 mm and 1.9±0.9 mm, respectively from left to right, and 8.3 mm and 4.9±1.9 mm, respectively from the anterior to the posterior direction. The maximum and the average rotational movement of the liver tumor were measured to be 3.3o and 2.6±1.3o, respectively for X (Left-Right) axis rotation, 4.8o and 2.3±1.0o, respectively for Y (Cranio-Caudal) axis rotation, 3.9o and 2.8±1.1o, respectively for Z (Anterior-Posterior) axis rotation. In addition, the average correlation error, which represents the treatment's accuracy was 1.1±0.7 mm. Conclusion

Species identification of Cannabis sativa using real-time quantitative PCR (qPCR).

Science.gov (United States)

Johnson, Christopher E; Premasuthan, Amritha; Satkoski Trask, Jessica; Kanthaswamy, Sree

2013-03-01

Most narcotics-related cases in the United States involve Cannabis sativa. Material is typically identified based on the cystolithic hairs on the leaves and with chemical tests to identify of the presence of cannabinoids. Suspect seeds are germinated into a viable plant so that morphological and chemical tests can be conducted. Seed germination, however, causes undue analytical delays. DNA analyses that involve the chloroplast and nuclear genomes have been developed for identification of C. sativa materials, but they require several nanograms of template DNA. Using the trnL 3' exon-trnF intragenic spacer regions within the C. sativa chloroplast, we have developed a real-time quantitative PCR assay that is capable of identifying picogram amounts of chloroplast DNA for species determination of suspected C. sativa material. This assay provides forensic science laboratories with a quick and reliable method to identify an unknown sample as C. sativa. © 2013 American Academy of Forensic Sciences.

A Human Fecal Contamination Score for Ranking Recreational Sites using the HF183/BacR287 Quantitative Real-Time PCR Method

Science.gov (United States)

Human fecal pollution of recreational waters remains a public health concern worldwide. As a result, there is a growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for water quality research and manag...

Species-specific detection and quantification of common barnacle larvae from the Japanese coast using quantitative real-time PCR.

Science.gov (United States)

Endo, Noriyuki; Sato, Kana; Matsumura, Kiyotaka; Yoshimura, Erina; Odaka, Yukiko; Nogata, Yasuyuki

2010-11-01

Species-specific detection and quantification methods for barnacle larvae using quantitative real-time polymerase chain reaction (qPCR) were developed. Species-specific primers for qPCR were designed for 13 barnacle species in the mitochondrial 12S ribosomal RNA gene region. Primer specificity was examined by PCR using template DNA extracted from each of the 13 barnacle species, other unidentified barnacle species, and field collected zooplankton samples. The resulting PCR products comprised single bands following agarose gel electrophoresis when the templates corresponded to primers. The amplifications were highly species-specific even for the field plankton samples. The field plankton samples were subjected to qPCR assay. The calculated DNA contents for each barnacle species were closely correlated with the number of larvae measured by microscopic examination. The method could be applied to quantify barnacle larvae in natural plankton samples.

Comparison of three-way and four-way calibration for the real-time quantitative analysis of drug hydrolysis in complex dynamic samples by excitation-emission matrix fluorescence

Science.gov (United States)

Yin, Xiao-Li; Gu, Hui-Wen; Liu, Xiao-Lu; Zhang, Shan-Hui; Wu, Hai-Long

2018-03-01

Multiway calibration in combination with spectroscopic technique is an attractive tool for online or real-time monitoring of target analyte(s) in complex samples. However, how to choose a suitable multiway calibration method for the resolution of spectroscopic-kinetic data is a troubling problem in practical application. In this work, for the first time, three-way and four-way fluorescence-kinetic data arrays were generated during the real-time monitoring of the hydrolysis of irinotecan (CPT-11) in human plasma by excitation-emission matrix fluorescence. Alternating normalization-weighted error (ANWE) and alternating penalty trilinear decomposition (APTLD) were used as three-way calibration for the decomposition of the three-way kinetic data array, whereas alternating weighted residual constraint quadrilinear decomposition (AWRCQLD) and alternating penalty quadrilinear decomposition (APQLD) were applied as four-way calibration to the four-way kinetic data array. The quantitative results of the two kinds of calibration models were fully compared from the perspective of predicted real-time concentrations, spiked recoveries of initial concentration, and analytical figures of merit. The comparison study demonstrated that both three-way and four-way calibration models could achieve real-time quantitative analysis of the hydrolysis of CPT-11 in human plasma under certain conditions. However, it was also found that both of them possess some critical advantages and shortcomings during the process of dynamic analysis. The conclusions obtained in this paper can provide some helpful guidance for the reasonable selection of multiway calibration models to achieve the real-time quantitative analysis of target analyte(s) in complex dynamic systems.

Validation of Reference Genes for Quantitative Expression Analysis by Real-Time RT-PCR in Four Lepidopteran Insects

OpenAIRE

Teng, Xiaolu; Zhang, Zan; He, Guiling; Yang, Liwen; Li, Fei

2012-01-01

Quantitative real-time polymerase chain reaction (qPCR) is an efficient and widely used technique to monitor gene expression. Housekeeping genes (HKGs) are often empirically selected as the reference genes for data normalization. However, the suitability of HKGs used as the reference genes has been seldom validated. Here, six HKGs were chosen (actin A3, actin A1, GAPDH, G3PDH, E2F, rp49) in four lepidopteran insects Bombyx mori L. (Lepidoptera: Bombycidae), Plutella xylostella L. (Plutellidae...

Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods.

Science.gov (United States)

Edagawa, Akiko; Kimura, Akio; Kawabuchi-Kurata, Takako; Adachi, Shinichi; Furuhata, Katsunori; Miyamoto, Hiroshi

2015-10-19

We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR), and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%). Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%). In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8%) compared with real-time qPCR alone (46/68, 67.6%). Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1%) compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%). Legionella was not detected in the remaining six samples (6/68, 8.8%), irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

Detection of Ophiocordyceps sinensis in soil by quantitative real-time PCR.

Science.gov (United States)

Peng, Qingyun; Zhong, Xin; Lei, Wei; Zhang, Guren; Liu, Xin

2013-03-01

Ophiocordyceps sinensis, one of the best known entomopathogenic fungi in traditional Chinese medicine, parasitizes larvae of the moth genus Thitarodes, which lives in soil tunnels. However, little is known about the spatial distribution of O. sinensis in the soil. We established a protocol for DNA extraction, purification, and quantification of O. sinensis in soil with quantitative real-time PCR targeting the internal transcribed spacer region. The method was assessed using 34 soil samples from Tibet. No inhibitory effects in purified soil DNA extracts were detected. The standard curve method for absolute DNA quantification generated crossing point values that were strongly and linearly correlated to the log10 of the initial amount of O. sinensis genomic DNA (r(2) = 0.999) over 7 orders of magnitude (4 × 10(1) to 4 × 10(7) fg). The amplification efficiency and y-intercept value of the standard curve were 1.953 and 37.70, respectively. The amount of O. sinensis genomic DNA decreased with increasing soil depth and horizontal distance from a sclerotium (P protocol is rapid, specific, sensitive, and provides a powerful tool for quantification of O. sinensis from soil.

IMU-based Real-time Pose Measurement system for Anterior Pelvic Plane in Total Hip Replacement Surgeries.

Science.gov (United States)

Zhe Cao; Shaojie Su; Hao Tang; Yixin Zhou; Zhihua Wang; Hong Chen

2017-07-01

With the aging of population, the number of Total Hip Replacement Surgeries (THR) increased year by year. In THR, inaccurate position of the implanted prosthesis may lead to the failure of the operation. In order to reduce the failure rate and acquire the real-time pose of Anterior Pelvic Plane (APP), we propose a measurement system in this paper. The measurement system includes two parts: Initial Pose Measurement Instrument (IPMI) and Real-time Pose Measurement Instrument (RPMI). IPMI is used to acquire the initial pose of the APP, and RPMI is used to estimate the real-time pose of the APP. Both are composed of an Inertial Measurement Unit (IMU) and magnetometer sensors. To estimate the attitude of the measurement system, the Extended Kalman Filter (EKF) is adopted in this paper. The real-time pose of the APP could be acquired together with the algorithm designed in the paper. The experiment results show that the Root Mean Square Error (RMSE) is within 1.6 degrees, which meets the requirement of THR operations.

Clinical evaluation of a quantitative real time polymerase chain reaction assay for diagnosis of primary Epstein-Barr virus infection in children.

Science.gov (United States)

Pitetti, Raymond D; Laus, Stella; Wadowsky, Robert M

2003-08-01

Epstein-Barr virus (EBV) infectious mononucleosis is often diagnosed based on characteristic clinical features and either a positive heterophil antibody test or serology, both of which can be unreliable in young children. Real time quantitative PCR assays that measure EBV DNA load in serum or plasma are highly sensitive in young children, but serum and plasma contain inhibitors of PCR which must be removed by DNA extraction techniques. A real time TaqMan PCR assay was designed and evaluated for simultaneously measuring EBV DNA load and validating the removal of PCR inhibitors from serum samples. A serum sample was available from patients classified serologically as primary EBV infection (n = 28), EBV-seronegative (n = 25) and EBV-seropositive (n = 26). Patients were classified as having EBV infectious mononucleosis if they had specified clinical findings and > or =10% atypical lymphocytes in peripheral blood or had a positive Monospot test result. DNA was purified by a spin column method and tested in PCR reactions with primers for EBV DNA polymerase gene and internal control targets. Amplification of the two PCR products was measured in real time with separate TaqMan DNA probes labeled with various fluorescent reporters. The mean age of study patients was 9 years, 4 months. Twenty-one (75%) of the patients in the primary EBV infection group, one (4%) of the seronegatives and none of the seropositives had detectable EBV DNA. Within the primary infection group, those with detectable virus were more likely than those without detectable virus to have evidence of lymphadenopathy (14 of 16 vs.1 of 5; P = 0.011), higher mean atypical (11.7 vs.0.9%; P = 0.002) and absolute atypical (1.5 vs.0.1 x 109/l; P = 0.004) lymphocyte count, higher mean absolute lymphocyte count (4.7 vs.2.3 x 109/l; P = 0.026) and higher mean aspartate aminotransferase value (119.8 vs.37.3 IU/l; P = 0.036). Ten patients, all in the primary infection group, had EBV infectious mononucleosis, and all

Quantitative Evaluation of Fire and EMS Mobilization Times

CERN Document Server

Upson, Robert

2010-01-01

Quantitative Evaluation of Fire and EMS Mobilization Times presents comprehensive empirical data on fire emergency and EMS call processing and turnout times, and aims to improve the operational benchmarks of NFPA peer consensus standards through a close examination of real-world data. The book also identifies and analyzes the elements that can influence EMS mobilization response times. Quantitative Evaluation of Fire and EMS Mobilization Times is intended for practitioners as a tool for analyzing fire emergency response times and developing methods for improving them. Researchers working in a

Real-time measurements and their effects on state estimation of distribution power system

DEFF Research Database (Denmark)

Han, Xue; You, Shi; Thordarson, Fannar

2013-01-01

between the estimated values (voltage and injected power) and the measurements are applied to evaluate the accuracy of the estimated grid states. Eventually, some suggestions are provided for the distribution grid operators on placing the real-time meters in the distribution grid.......This paper aims at analyzing the potential value of using different real-time metering and measuring instruments applied in the low voltage distribution networks for state-estimation. An algorithm is presented to evaluate different combinations of metering data using a tailored state estimator....... It is followed by a case study based on the proposed algorithm. A real distribution grid feeder with different types of meters installed either in the cabinets or at the customer side is selected for simulation and analysis. Standard load templates are used to initiate the state estimation. The deviations...

Visually estimated ejection fraction by two dimensional and triplane echocardiography is closely correlated with quantitative ejection fraction by real-time three dimensional echocardiography.

Science.gov (United States)

Shahgaldi, Kambiz; Gudmundsson, Petri; Manouras, Aristomenis; Brodin, Lars-Ake; Winter, Reidar

2009-08-25

Visual assessment of left ventricular ejection fraction (LVEF) is often used in clinical routine despite general recommendations to use quantitative biplane Simpsons (BPS) measurements. Even thou quantitative methods are well validated and from many reasons preferable, the feasibility of visual assessment (eyeballing) is superior. There is to date only sparse data comparing visual EF assessment in comparison to quantitative methods available. The aim of this study was to compare visual EF assessment by two-dimensional echocardiography (2DE) and triplane echocardiography (TPE) using quantitative real-time three-dimensional echocardiography (RT3DE) as the reference method. Thirty patients were enrolled in the study. Eyeballing EF was assessed using apical 4-and 2 chamber views and TP mode by two experienced readers blinded to all clinical data. The measurements were compared to quantitative RT3DE. There were an excellent correlation between eyeballing EF by 2D and TP vs 3DE (r = 0.91 and 0.95 respectively) without any significant bias (-0.5 +/- 3.7% and -0.2 +/- 2.9% respectively). Intraobserver variability was 3.8% for eyeballing 2DE, 3.2% for eyeballing TP and 2.3% for quantitative 3D-EF. Interobserver variability was 7.5% for eyeballing 2D and 8.4% for eyeballing TP. Visual estimation of LVEF both using 2D and TP by an experienced reader correlates well with quantitative EF determined by RT3DE. There is an apparent trend towards a smaller variability using TP in comparison to 2D, this was however not statistically significant.

Real time NO emissions measurement during cold start in LPG SI engine

International Nuclear Information System (INIS)

Li, Gong; Liu, Zhimin; Li, Zhilong; Qiu, Dongping; Li, Liguang

2007-01-01

To identify combustion occurrence is very important. Traditionally, cylinder pressure has been used as a criterion of combustion occurrence, but it can be unreliable when identifying lean mixture combustion (there is little difference in the cylinder pressure trace between the firing cycle and motoring cycles at the lean combustion limit). This is particularly important for fuels like LPG, which have a good capacity for lean combustion. In this study, a fast response NO detector, CambustionfNOx400, based on the chemiluminescence method, was used to measure real time NO emissions in order to evaluate the technique as a criterion for establishing combustion occurrence. At the same time, this paper presents an investigation of the characteristics of real time NO emissions of the first firing cycle during cold start in a LPG SI engine to determine the optimal excess air factor of the first firing cycle, and the cylinder pressure and crank shaft speed of the engine were measured and recorded. Test results show that the excess air ratio directly influences the cylinder pressure, engine speed and NO emissions of the first firing cycle. As the excess air coefficient is reduced from the lean misfiring limit, NO emissions increase quickly, then reduce quickly and then reduce slowly. NO emissions generally increase with peak cylinder pressure, even at constant excess air coefficient. Real time NO emissions can be used to identify cylinder combustion and misfire occurrence during engine cranking, even at the dilute combustion limit, and real time NO emission can be used to understand the combustion and misfire occurrence. (author)

Statistical aspects of quantitative real-time PCR experiment design

Czech Academy of Sciences Publication Activity Database

Kitchen, R.R.; Kubista, Mikael; Tichopád, Aleš

2010-01-01

Roč. 50, č. 4 (2010), s. 231-236 ISSN 1046-2023 R&D Projects: GA AV ČR IAA500520809 Institutional research plan: CEZ:AV0Z50520701 Keywords : Real-time PCR * Experiment design * Nested analysis of variance Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.527, year: 2010

Method and apparatus for real-time measurement of fuel gas compositions and heating values

Science.gov (United States)

Zelepouga, Serguei; Pratapas, John M.; Saveliev, Alexei V.; Jangale, Vilas V.

2016-03-22

An exemplary embodiment can be an apparatus for real-time, in situ measurement of gas compositions and heating values. The apparatus includes a near infrared sensor for measuring concentrations of hydrocarbons and carbon dioxide, a mid infrared sensor for measuring concentrations of carbon monoxide and a semiconductor based sensor for measuring concentrations of hydrogen gas. A data processor having a computer program for reducing the effects of cross-sensitivities of the sensors to components other than target components of the sensors is also included. Also provided are corresponding or associated methods for real-time, in situ determination of a composition and heating value of a fuel gas.

A real-time assessment of measurement uncertainty in the experimental characterization of sprays

International Nuclear Information System (INIS)

Panão, M R O; Moreira, A L N

2008-01-01

This work addresses the estimation of the measurement uncertainty of discrete probability distributions used in the characterization of sprays. A real-time assessment of this measurement uncertainty is further investigated, particularly concerning the informative quality of the measured distribution and the influence of acquiring additional information on the knowledge retrieved from statistical analysis. The informative quality is associated with the entropy concept as understood in information theory (Shannon entropy), normalized by the entropy of the most informative experiment. A new empirical correlation is derived between the error accuracy of a discrete cumulative probability distribution and the normalized Shannon entropy. The results include case studies using: (i) spray impingement measurements to study the applicability of the real-time assessment of measurement uncertainty, and (ii) the simulation of discrete probability distributions of unknown shape or function to test the applicability of the new correlation

Digital video timing analyzer for the evaluation of PC-based real-time simulation systems

Science.gov (United States)

Jones, Shawn R.; Crosby, Jay L.; Terry, John E., Jr.

2009-05-01

Due to the rapid acceleration in technology and the drop in costs, the use of commercial off-the-shelf (COTS) PC-based hardware and software components for digital and hardware-in-the-loop (HWIL) simulations has increased. However, the increase in PC-based components creates new challenges for HWIL test facilities such as cost-effective hardware and software selection, system configuration and integration, performance testing, and simulation verification/validation. This paper will discuss how the Digital Video Timing Analyzer (DiViTA) installed in the Aviation and Missile Research, Development and Engineering Center (AMRDEC) provides quantitative characterization data for PC-based real-time scene generation systems. An overview of the DiViTA is provided followed by details on measurement techniques, applications, and real-world examples of system benefits.

Comparison of three-way and four-way calibration for the real-time quantitative analysis of drug hydrolysis in complex dynamic samples by excitation-emission matrix fluorescence.

Science.gov (United States)

Yin, Xiao-Li; Gu, Hui-Wen; Liu, Xiao-Lu; Zhang, Shan-Hui; Wu, Hai-Long

2018-03-05

Multiway calibration in combination with spectroscopic technique is an attractive tool for online or real-time monitoring of target analyte(s) in complex samples. However, how to choose a suitable multiway calibration method for the resolution of spectroscopic-kinetic data is a troubling problem in practical application. In this work, for the first time, three-way and four-way fluorescence-kinetic data arrays were generated during the real-time monitoring of the hydrolysis of irinotecan (CPT-11) in human plasma by excitation-emission matrix fluorescence. Alternating normalization-weighted error (ANWE) and alternating penalty trilinear decomposition (APTLD) were used as three-way calibration for the decomposition of the three-way kinetic data array, whereas alternating weighted residual constraint quadrilinear decomposition (AWRCQLD) and alternating penalty quadrilinear decomposition (APQLD) were applied as four-way calibration to the four-way kinetic data array. The quantitative results of the two kinds of calibration models were fully compared from the perspective of predicted real-time concentrations, spiked recoveries of initial concentration, and analytical figures of merit. The comparison study demonstrated that both three-way and four-way calibration models could achieve real-time quantitative analysis of the hydrolysis of CPT-11 in human plasma under certain conditions. However, it was also found that both of them possess some critical advantages and shortcomings during the process of dynamic analysis. The conclusions obtained in this paper can provide some helpful guidance for the reasonable selection of multiway calibration models to achieve the real-time quantitative analysis of target analyte(s) in complex dynamic systems. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparative study of β-glucan induced respiratory burst measured by nitroblue tetrazolium assay and real-time luminol-enhanced chemiluminescence assay in common carp (Cyprinus carpio L.).

Science.gov (United States)

Vera-Jimenez, N I; Pietretti, D; Wiegertjes, G F; Nielsen, M E

2013-05-01

The respiratory burst is an important feature of the immune system. The increase in cellular oxygen uptake that marks the initiation of the respiratory burst is followed by the production of reactive oxygen species (ROS) such as superoxide anion and hydrogen peroxide which plays a role in the clearance of pathogens and tissue regeneration processes. Therefore, the respiratory burst and associated ROS constitute important indicators of fish health status. This paper compares two methods for quantitation of ROS produced during the respiratory burst in common carp: the widely used, single-point measurement based on the intracellular reduction of nitroblue tetrazolium (NBT) and a real-time luminol-enhanced assay based on the detection of native chemiluminescence. Both assays allowed for detection of dose-dependent changes in magnitude of the respiratory burst response induced by β-glucans in head kidney cells of carp. However, whereas the NBT assay was shown to detect the production of only superoxide anions, the real-time luminol-enhanced assay could detect the production of both superoxide anions and hydrogen peroxide. Only the chemiluminescence assay could reliably record the production of ROS on a real-time scale at frequent and continual time intervals for time course experiments, providing more detailed information on the respiratory burst response. The real-time chemiluminescence assay was used to measure respiratory burst activity in macrophage and neutrophilic granulocyte-enriched head kidney cell fractions and total head kidney cell suspensions and proved to be a fast, reliable, automated multiwell microplate assay to quantitate fish health status modulated by β-glucans. Copyright © 2013 Elsevier Ltd. All rights reserved.

Fast real-time polymerase chain reaction for quantitative detection of Lactobacillus delbrueckii bacteriophages in milk.

Science.gov (United States)

Martín, Maria Cruz; del Rio, Beatriz; Martínez, Noelia; Magadán, Alfonso H; Alvarez, Miguel A

2008-12-01

One of the main microbiological problems of the dairy industry is the susceptibility of starter bacteria to virus infections. Lactobacillus delbrueckii, a component of thermophilic starter cultures used in the manufacture of several fermented dairy products, including yogurt, is also sensitive to bacteriophage attacks. To avoid the problems associated with these viruses, quick and sensitive detection methods are necessary. In the present study, a fast real-time quantitative polymerase chain reaction assay for the direct detection and quantification of L. delbrueckii phages in milk was developed. A set of primers and a TaqMan MGB probe was designed, based on the lysin gene sequence of different L. delbrueckii phages. The results show the proposed method to be a rapid (total processing time 30 min), specific and highly sensitive technique for detecting L. delbrueckii phages in milk.

Real-Time Aerodynamic Parameter Estimation without Air Flow Angle Measurements

Science.gov (United States)

Morelli, Eugene A.

2010-01-01

A technique for estimating aerodynamic parameters in real time from flight data without air flow angle measurements is described and demonstrated. The method is applied to simulated F-16 data, and to flight data from a subscale jet transport aircraft. Modeling results obtained with the new approach using flight data without air flow angle measurements were compared to modeling results computed conventionally using flight data that included air flow angle measurements. Comparisons demonstrated that the new technique can provide accurate aerodynamic modeling results without air flow angle measurements, which are often difficult and expensive to obtain. Implications for efficient flight testing and flight safety are discussed.

Investigations on abundance and activity of microbial sponge symbionts using quantitative real - time PCR

DEFF Research Database (Denmark)

Kumala, Lars; Hentschel, Ute; Bayer, Kristina

Marine sponges are hosts to dense and diverse microbial consortia that are likely to play a key role in the metabolic processes of the host sponge due to their enormous abundance. Common symbioses between nitrogen transforming microorganisms and sponges indicate complex nitrogen cycling within...... the host. Of particular interest is determining the community structure and function of microbial symbionts in order to gain deeper insight into host-symbiont interactions. We investigated the abundance and activity of microbial symbionts in two Mediterranean sponge species using quantitative real-time PCR....... An absolute quantification of functional genes and transcripts in archaeal and bacterial symbionts was conducted to determine their involvement in nitrification and denitrification, comparing the low microbial abundance (LMA) sponge Dysidea avara with the high microbial abundance (HMA) representative Aplysina...

Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods

Directory of Open Access Journals (Sweden)

Akiko Edagawa

2015-10-01

Full Text Available We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR, and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%. Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%. In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8% compared with real-time qPCR alone (46/68, 67.6%. Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1% compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%. Legionella was not detected in the remaining six samples (6/68, 8.8%, irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

Selection of Suitable Reference Genes for Quantitative Real-time PCR in Sapium sebiferum

Directory of Open Access Journals (Sweden)

Xue Chen

2017-05-01

Full Text Available Chinese tallow (Sapium sebiferum L. is a promising landscape and bioenergy plant. Measuring gene expression by quantitative real-time polymerase chain reaction (qRT-PCR can provide valuable information on gene function. Stably expressed reference genes for normalization are a prerequisite for ensuring the accuracy of the target gene expression level among different samples. However, the reference genes in Chinese tallow have not been systematically validated. In this study, 12 candidate reference genes (18S, GAPDH, UBQ, RPS15, SAND, TIP41, 60S, ACT7, PDF2, APT, TBP, and TUB were investigated with qRT-PCR in 18 samples, including those from different tissues, from plants treated with sucrose and cold stresses. The data were calculated with four common algorithms, geNorm, BestKeeper, NormFinder, and the delta cycle threshold (ΔCt. TIP41 and GAPDH were the most stable for the tissue-specific experiment, GAPDH and 60S for cold treatment, and GAPDH and UBQ for sucrose stresses, while the least stable genes were 60S, TIP41, and 18S respectively. The comprehensive results showed APT, GAPDH, and UBQ to be the top-ranked stable genes across all the samples. The stability of 60S was the lowest during all experiments. These selected reference genes were further validated by comparing the expression profiles of the chalcone synthase gene in Chinese tallow in different samples. The results will help to improve the accuracy of gene expression studies in Chinese tallow.

Identification of three novel OA1 gene mutations identified in three families misdiagnosed with congenital nystagmus and carrier status determination by real-time quantitative PCR assay

Directory of Open Access Journals (Sweden)

Hamel Christian

2003-01-01

Full Text Available Abstract Background X-linked ocular albinism type 1 (OA1 is caused by mutations in OA1 gene, which encodes a membrane glycoprotein localised to melanosomes. OA1 mainly affects pigment production in the eye, resulting in optic changes associated with albinism including hypopigmentation of the retina, nystagmus, strabismus, foveal hypoplasia, abnormal crossing of the optic fibers and reduced visual acuity. Affected Caucasian males usually appear to have normal skin and hair pigment. Results We identified three previously undescribed mutations consisting of two intragenic deletions (one encompassing exon 6, the other encompassing exons 7–8, and a point mutation (310delG in exon 2. We report the development of a new method for diagnosis of heterozygous deletions in OA1 gene based on measurement of gene copy number using real-time quantitative PCR from genomic DNA. Conclusion The identification of OA1 mutations in families earlier reported as families with hereditary nystagmus indicate that ocular albinism type 1 is probably underdiagnosed. Our method of real-time quantitative PCR of OA1 exons with DMD exon as external standard performed on the LightCycler™ allows quick and accurate carrier-status assessment for at-risk females.

Accuracy of real time radiography burning rate measurement

Science.gov (United States)

Olaniyi, Bisola

The design of a solid propellant rocket motor requires the determination of a propellant's burning-rate and its dependency upon environmental parameters. The requirement that the burning-rate be physically measured, establishes the need for methods and equipment to obtain such data. A literature review reveals that no measurement has provided the desired burning rate accuracy. In the current study, flash x-ray modeling and digitized film-density data were employed to predict motor-port area to length ratio. The pre-fired port-areas and base burning rate were within 2.5% and 1.2% of their known values, respectively. To verify the accuracy of the method, a continuous x-ray and a solid propellant rocket motor model (Plexiglas cylinder) were used. The solid propellant motor model was translated laterally through a real-time radiography system at different speeds simulating different burning rates. X-ray images were captured and the burning-rate was then determined. The measured burning rate was within 1.65% of the known values.

A quantitative method to evaluate mesenchymal stem cell lipofection using real-time PCR.

Science.gov (United States)

Ribeiro, S C; Mendes, R; Madeira, C; Monteiro, G A; da Silva, C L; Cabral, J M S

2010-01-01

Genetic modification of human mesenchymal stem cells (MSC) is a powerful tool to improve the therapeutic utility of these cells and to increase the knowledge on their regulation mechanisms. In this context, strong efforts have been made recently to develop efficient nonviral gene delivery systems. Although several studies addressed this question most of them use the end product of a reporter gene instead of the DNA uptake quantification to test the transfection efficiency. In this study, we established a method based on quantitative real-time PCR (RT-PCR) to determine the intracellular plasmid DNA copy number in human MSC after lipofection. The procedure requires neither specific cell lysis nor DNA purification. The influence of cell number on the RT-PCR sensitivity was evaluated. The method showed good reproducibility, high sensitivity, and a wide linear range of 75-2.5 x 10⁶ plasmid DNA copies per cell. RT-PCR results were then compared with the percentage of transfected cells assessed by flow cytometry analysis, which showed that flow cytometry-based results are not always proportional to plasmid cellular uptake determined by RT-PCR. This work contributed for the establishment of a rapid quantitative assay to determine intracellular plasmid DNA in stem cells, which will be extremely beneficial for the optimization of gene delivery strategies. © 2010 American Institute of Chemical Engineers

QUANTITATION OF INTRACELLULAR NAD(P)H IN LIVING CELLS CAN MONITOR AN IMBALANCE OF DNA SINGLE STRAND BREAK REPAIR IN REAL TIME

Science.gov (United States)

Quantitation of intracellular NAD(P)H in living cells can monitor an imbalance of DNA single strand break repair in real time.ABSTRACTDNA single strand breaks (SSBs) are one of the most frequent DNA lesions in genomic DNA generated either by oxidative stress or du...

Real-Time Measurement of Host Bioenergetics During Mycobacterium Tuberculosis Infection

Science.gov (United States)

2015-05-01

AWARD NUMBER: W81XWH-13-1-0149 TITLE: “Real-Time Measurement of Host Bioenergetics During Mycobacterium Tuberculosis Infection...successfully adapted metabolic flux analysis using a Seahorse XF96 metabolic flux analyzer to study Mycobacterium tuberculosis energy metabolism in an...Mycobacterium tuberculosis function. In: Systems Biology of Tuberculosis . Editors: J McFadden, D Beste and A Kierzek. 2013. Springer, New York, NY. 2

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...

Visually estimated ejection fraction by two dimensional and triplane echocardiography is closely correlated with quantitative ejection fraction by real-time three dimensional echocardiography

Directory of Open Access Journals (Sweden)

Manouras Aristomenis

2009-08-01

Full Text Available Abstract Background Visual assessment of left ventricular ejection fraction (LVEF is often used in clinical routine despite general recommendations to use quantitative biplane Simpsons (BPS measurements. Even thou quantitative methods are well validated and from many reasons preferable, the feasibility of visual assessment (eyeballing is superior. There is to date only sparse data comparing visual EF assessment in comparison to quantitative methods available. The aim of this study was to compare visual EF assessment by two-dimensional echocardiography (2DE and triplane echocardiography (TPE using quantitative real-time three-dimensional echocardiography (RT3DE as the reference method. Methods Thirty patients were enrolled in the study. Eyeballing EF was assessed using apical 4-and 2 chamber views and TP mode by two experienced readers blinded to all clinical data. The measurements were compared to quantitative RT3DE. Results There were an excellent correlation between eyeballing EF by 2D and TP vs 3DE (r = 0.91 and 0.95 respectively without any significant bias (-0.5 ± 3.7% and -0.2 ± 2.9% respectively. Intraobserver variability was 3.8% for eyeballing 2DE, 3.2% for eyeballing TP and 2.3% for quantitative 3D-EF. Interobserver variability was 7.5% for eyeballing 2D and 8.4% for eyeballing TP. Conclusion Visual estimation of LVEF both using 2D and TP by an experienced reader correlates well with quantitative EF determined by RT3DE. There is an apparent trend towards a smaller variability using TP in comparison to 2D, this was however not statistically significant.

Real-time individualized training vectors for experiential learning.

Energy Technology Data Exchange (ETDEWEB)

Willis, Matt; Tucker, Eilish Marie; Raybourn, Elaine Marie; Glickman, Matthew R.; Fabian, Nathan

2011-01-01

Military training utilizing serious games or virtual worlds potentially generate data that can be mined to better understand how trainees learn in experiential exercises. Few data mining approaches for deployed military training games exist. Opportunities exist to collect and analyze these data, as well as to construct a full-history learner model. Outcomes discussed in the present document include results from a quasi-experimental research study on military game-based experiential learning, the deployment of an online game for training evidence collection, and results from a proof-of-concept pilot study on the development of individualized training vectors. This Lab Directed Research & Development (LDRD) project leveraged products within projects, such as Titan (Network Grand Challenge), Real-Time Feedback and Evaluation System, (America's Army Adaptive Thinking and Leadership, DARWARS Ambush! NK), and Dynamic Bayesian Networks to investigate whether machine learning capabilities could perform real-time, in-game similarity vectors of learner performance, toward adaptation of content delivery, and quantitative measurement of experiential learning.

Real-time measurement and control at JET signal processing and physics analysis for diagnostics

International Nuclear Information System (INIS)

Felton, R.; Joffrin, E.; Murari, A.

2005-01-01

To meet the requirements of the scientific programme, the EFDA JET real-time measurement and control project has developed an integrated set of real-time plasma measurements, experiment control and communication facilities. Traditional experiments collected instrument data during the plasma pulse and calculated physics data after the pulse. The challenge for continuous tokamak operation is to calculate the physics data in real-time, keeping up with the evolution of the plasma. In JET, many plasma diagnostics have been augmented with extra data acquisition and signal-processing systems so that they can both capture instrument data for conventional post-pulse analysis and calculate calibrated, validated physics results in real-time. During the pulse, the systems send sampled data sets into a network, which distributes the data to several destinations. The receiving systems may do further analysis, integrating data from several measurements, or may control the plasma scenario by heating or fuelling. The simplest real-time diagnostic systems apply scale factors to the signals, as with the electron cyclotron emission (ECE) diagnostic's 96 tuned radiometer channels, giving the electron temperature profile. In various spectroscopy diagnostics, spectral features are least-squares-fitted to measure spectra from several lines of sight, within 50 ms. Ion temperatures and rotation speed can be calculated from the line widths and shifts. For diagnostics using modulation techniques, the systems implement digital-signal processing phase trackers, lock-in amplifiers and filters, e.g., the far infrared (FIR) interferometer samples 15 channels at 400 kHz for 30 s, i.e., six million samples per second. Diagnostics have specific lines of sight, spatial channels, and various sampling rates. The heating/fuelling systems have relatively coarse spatial localisation. Analysis systems have been developed to integrate the basic physics data into smaller sets of controllable parameters on a

Real-Time Inhibitor Recession Measurements in Two Space Shuttle Reusable Solid Rocket Motors

Science.gov (United States)

McWhorter, B. B.; Ewing, M. E.; Bolton, D. E.; Albrechtsen, K. U.; Earnest, T. E.; Noble, T. C.; Longaker, M.

2003-01-01

Real-time internal motor insulation char line recession measurements have been evaluated for two full-scale static tests of the Space Shuttle Reusable Solid Rocket Motor (RSRM). These char line recession measurements were recorded on the forward facing propellant grain inhibitors to better understand the thermal performance of these inhibitors. The RSRM propellant grain inhibitors are designed to erode away during motor operation, thus making it difficult to use post-fire observations to determine inhibitor thermal performance. Therefore, this new internal motor instrumentation is invaluable in establishing an accurate understanding of inhibitor recession versus motor operation time. The data for the first test was presented at the 37th AIAA/ASME/SAE/ASEE Joint Propulsion Conference and Exhibit (AIAA 2001-3280) in July 2001. Since that time, a second full scale static test has delivered additional real-time data on inhibitor thermal performance. The evaluation of this data is presented in this paper. The second static test, in contrast to the first test, used a slightly different arrangement of instrumentation in the inhibitors. This instrumentation has yielded a better understanding of the inhibitor time dependent inboard tip recession. Graphs of inhibitor recession profiles with time are presented. Inhibitor thermal ablation models have been created from theoretical principals. The model predictions compare favorably with data from both tests. This verified modeling effort is important to support new inhibitor designs for a five segment Space Shuttle solid rocket motor. The internal instrumentation project on RSRM static tests is providing unique opportunities for other real-time internal motor measurements that could not otherwise be directly quantified.

[Usefulness of a real-time quantitative polymerase-chain reaction (PCR) assay for the diagnosis of congenital and postnatal cytomegalovirus infection].

Science.gov (United States)

Reina, J; Weber, I; Riera, E; Busquets, M; Morales, C

2014-05-01

Cytomegalovirus (CMV) is the main virus causing congenital and postnatal infections in the pediatric population. The aim of this study is to evaluate the usefulness of a quantitative real-time PCR in the diagnosis of these infections using urine as a single sample. We studied all the urine samples of newborns (< 7 days) with suspected congenital infection, and urine of patients with suspected postnatal infection (urine negative at birth). Urines were simultaneously studied by cell culture, qualitative PCR (PCRc), and quantitative real-time PCR (PCRq). We analyzed 332 urine samples (270 to rule out congenital infection and 62 postnatal infections). Of the first, 22 were positive in the PCRq, 19 in the PCRc, and 17 in the culture. PCRq had a sensitivity of 100%, on comparing the culture with the rest of the techniques. Using the PCRq as a reference method, culture had a sensitivity of 77.2%, and PCRc 86.3%. In cases of postnatal infection, PCRq detected 16 positive urines, the PCRq 12, and the cell culture 10. The urines showed viral loads ranging from 2,178 to 116,641 copies/ml. The genomic amplification technique PCRq in real time was more sensitive than the other techniques evaluated. This technique should be considered as a reference (gold standard), leaving the cell culture as a second diagnostic level. The low cost and the automation of PCRq would enable the screening for CMV infection in large neonatal and postnatal populations. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

Detection of Haemophilus influenzae in respiratory secretions from pneumonia patients by quantitative real-time polymerase chain reaction.

Science.gov (United States)

Abdeldaim, Guma M K; Strålin, Kristoffer; Kirsebom, Leif A; Olcén, Per; Blomberg, Jonas; Herrmann, Björn

2009-08-01

A quantitative real-time polymerase chain reaction (PCR) based on the omp P6 gene was developed to detect Haemophilus influenzae. Its specificity was determined by analysis of 29 strains of 11 different Haemophilus spp. and was compared with PCR assays having other target genes: rnpB, 16S rRNA, and bexA. The method was evaluated on nasopharyngeal aspirates from 166 adult patients with community-acquired pneumonia. When 10(4) DNA copies/mL was used as cutoff limit for the method, P6 PCR had a sensitivity of 97.5% and a specificity of 96.0% compared with the culture. Of 20 culture-negative but P6 PCR-positive cases, 18 were confirmed by fucK PCR as H. influenzae. Five (5.9%) of 84 nasopharyngeal aspirates from adult controls tested PCR positive. We conclude that the P6 real-time PCR is both sensitive and specific for identification of H. influenzae in respiratory secretions. Quantification facilitates discrimination between disease-causing H. influenzae strains and commensal colonization.

Advances in LWD pressure measurements: smart, time optimized pretests and on demand real-time transmission applications

Energy Technology Data Exchange (ETDEWEB)

Serafim, Robson; Ferraris, Paolo [Schlumberger, Rio de Janeiro, RJ (Brazil)

2008-07-01

The StethoScope Logging While Drilling (LWD) Pressure Measurement, introduced in Brazil in 2005, has been extensively used in deep water environment to provide reservoir pressure and mobility in real-time. In the last three years the StethoScope service was further enhanced to allow better real time monitoring using a larger transmission rate, higher RT data resolution and remote visualization. In order to guarantee stable formation pressures with a limited test duration under a wide range of conditions, Time Optimized Pretests (TOP) were developed. These tests adjust automatically drawdown and buildup parameters as a function of formation characteristics (pressure/mobility) without requiring any input from the operator. On-demand frame (ODF), an advanced telemetry triggered automatically during the pressure tests, allowed to increase equivalent transmission rate and resolution and to include quality indices computed downhole. This paper is focused on the TOP and ODF Field Test results in Brazil, which proved to be useful and reliable options for better real-time decisions together with remote monitoring visualization implemented by the RTMonitor program. (author)

Comparative study of standard space and real space analysis of quantitative MR brain data.

Science.gov (United States)

Aribisala, Benjamin S; He, Jiabao; Blamire, Andrew M

2011-06-01

To compare the robustness of region of interest (ROI) analysis of magnetic resonance imaging (MRI) brain data in real space with analysis in standard space and to test the hypothesis that standard space image analysis introduces more partial volume effect errors compared to analysis of the same dataset in real space. Twenty healthy adults with no history or evidence of neurological diseases were recruited; high-resolution T(1)-weighted, quantitative T(1), and B(0) field-map measurements were collected. Algorithms were implemented to perform analysis in real and standard space and used to apply a simple standard ROI template to quantitative T(1) datasets. Regional relaxation values and histograms for both gray and white matter tissues classes were then extracted and compared. Regional mean T(1) values for both gray and white matter were significantly lower using real space compared to standard space analysis. Additionally, regional T(1) histograms were more compact in real space, with smaller right-sided tails indicating lower partial volume errors compared to standard space analysis. Standard space analysis of quantitative MRI brain data introduces more partial volume effect errors biasing the analysis of quantitative data compared to analysis of the same dataset in real space. Copyright © 2011 Wiley-Liss, Inc.

Development of a dual-tracer real-time particle dry-deposition measurement technique for simple and complex terrain

International Nuclear Information System (INIS)

Sehmel, G.A.; Hodgson, W.H.; Campbell, J.A.

1979-01-01

Detectors are being developed and tested for measuring the airborne concentrations of lithium particles and SF 6 gas in real time. The airborne lithium detector will be used for real-time measurements of both particle dry-deposition velocities and resuspension rates. Both the lithium and SF 6 detectors will be used for measuring dry deposition in field experiments

Real-time scintillation probe measurement of left ventricular function

International Nuclear Information System (INIS)

Green, M.V.; Ostrow, H.G.; Bacharach, S.L.; Allen, S.I.; Bonow, R.O.; Johnston, G.S.

1981-01-01

The micro-processor based system described in this report was designed for maximum flexibility and utility. While the principle function of the system is to acquire, create, analyze and display (in real-time) left ventricular time activity (or volume) curves, provision is also made to acquire additional physiologic signals (e.g., ECG, flowmeter, etc.) and to calculate and display relationships between these various data. The system was designed for interactive use so that the system user can alter the course of a series of measurements based on previous results. These general capabilities are illustrated with several examples. In the first, LV function was measured continuously in a subject from (supine) rest through exercise and recovery. The second example illustrates the use of the system in acquiring (LV) pressure-volume loops. Several technical problems, such as correction for LV background radiation, appear at present to limit the probes applicability. Even now, however, probe systems are demonstrably useful in the study of global left ventricular function when this function is changing rapidly with time in response to various interventions. (orig.) [de

Quantum measurement and real-time feedback with a spin-register in diamond

NARCIS (Netherlands)

Blok, M.S.

2015-01-01

Gaining precise control over quantum systems is crucial for applications in quantum information processing and quantum sensing and to perform experimental tests of quantum mechanics. The experiments presented in this thesis implement quantum measurements and real-time feedback protocols that can

Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay

Science.gov (United States)

Ogrean, Christy; Jackson, Ben; Covino, James

2010-01-01

The Solaris qPCR Gene Expression Assay is a novel type of primer/probe set, designed to simplify the qPCR process while maintaining the sensitivity and accuracy of the assay. These primer/probe sets are pre-designed to >98% of the human and mouse genomes and feature significant improvements from previously available technologies. These improvements were made possible by virtue of a novel design algorithm, developed by Thermo Scientific bioinformatics experts. Several convenient features have been incorporated into the Solaris qPCR Assay to streamline the process of performing quantitative real-time PCR. First, the protocol is similar to commonly employed alternatives, so the methods used during qPCR are likely to be familiar. Second, the master mix is blue, which makes setting the qPCR reactions easier to track. Third, the thermal cycling conditions are the same for all assays (genes), making it possible to run many samples at a time and reducing the potential for error. Finally, the probe and primer sequence information are provided, simplifying the publication process. Here, we demonstrate how to obtain the appropriate Solaris reagents using the GENEius product search feature found on the ordering web site (www.thermo.com/solaris) and how to use the Solaris reagents for performing qPCR using the standard curve method. PMID:20567213

Protein Analysis Using Real-Time PCR Instrumentation: Incorporation in an Integrated, Inquiry-Based Project

Science.gov (United States)

Southard, Jonathan N.

2014-01-01

Instrumentation for real-time PCR is used primarily for amplification and quantitation of nucleic acids. The capability to measure fluorescence while controlling temperature in multiple samples can also be applied to the analysis of proteins. Conformational stability and changes in stability due to ligand binding are easily assessed. Protein…

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...

Portable Dew Point Mass Spectrometry System for Real-Time Gas and Moisture Analysis

Science.gov (United States)

Arkin, C.; Gillespie, Stacey; Ratzel, Christopher

2010-01-01

A portable instrument incorporates both mass spectrometry and dew point measurement to provide real-time, quantitative gas measurements of helium, nitrogen, oxygen, argon, and carbon dioxide, along with real-time, quantitative moisture analysis. The Portable Dew Point Mass Spectrometry (PDP-MS) system comprises a single quadrupole mass spectrometer and a high vacuum system consisting of a turbopump and a diaphragm-backing pump. A capacitive membrane dew point sensor was placed upstream of the MS, but still within the pressure-flow control pneumatic region. Pressure-flow control was achieved with an upstream precision metering valve, a capacitance diaphragm gauge, and a downstream mass flow controller. User configurable LabVIEW software was developed to provide real-time concentration data for the MS, dew point monitor, and sample delivery system pressure control, pressure and flow monitoring, and recording. The system has been designed to include in situ, NIST-traceable calibration. Certain sample tubing retains sufficient water that even if the sample is dry, the sample tube will desorb water to an amount resulting in moisture concentration errors up to 500 ppm for as long as 10 minutes. It was determined that Bev-A-Line IV was the best sample line to use. As a result of this issue, it is prudent to add a high-level humidity sensor to PDP-MS so such events can be prevented in the future.

Using RADFET for the real-time measurement of gamma radiation dose rate

Science.gov (United States)

Andjelković, Marko S.; Ristić, Goran S.; Jakšić, Aleksandar B.

2015-02-01

RADFETs (RADiation sensitive Field Effect Transistors) are integrating ionizing radiation dosimeters operating on the principle of conversion of radiation-induced threshold voltage shift into absorbed dose. However, one of the major drawbacks of RADFETs is the inability to provide the information on the dose rate in real-time using the conventional absorbed dose measurement technique. The real-time monitoring of dose rate and absorbed dose can be achieved with the current mode dosimeters such as PN and PIN diodes/photodiodes, but these dosimeters have some limitations as absorbed dose meters and hence they are often not a suitable replacement for RADFETs. In that sense, this paper investigates the possibility of using the RADFET as a real-time dose rate meter so that it could be applied for simultaneous online measurement of the dose rate and absorbed dose. A RADFET sample, manufactured by Tyndall National Institute, Cork, Ireland, was tested as a dose rate meter under gamma irradiation from a Co-60 source. The RADFET was configured as a PN junction, such that the drain, gate and source terminals were grounded, while the radiation-induced current was measured at the bulk terminal, whereby the bulk was successively biased with 0 , 10 , 20  and 30 V. In zero-bias mode the radiation-induced current was unstable, but in the biased mode the current response was stable for the investigated dose rates from 0.65  to 32.1 Gy h-1 and up to the total absorbed dose of 25 Gy. The current increased with the dose rate in accordance with the power law, whereas the sensitivity of the current read-out was linear with respect to the applied bias voltage. Comparison with previously analyzed PIN photodiodes has shown that the investigated RADFET is competitive with PIN photodiodes as a gamma radiation dose rate meter and therefore has the potential to be employed for the real-time monitoring of the dose rate and absorbed dose.

Using RADFET for the real-time measurement of gamma radiation dose rate

International Nuclear Information System (INIS)

Andjelković, Marko S; Ristić, Goran S; Jakšić, Aleksandar B

2015-01-01

RADFETs (RADiation sensitive Field Effect Transistors) are integrating ionizing radiation dosimeters operating on the principle of conversion of radiation-induced threshold voltage shift into absorbed dose. However, one of the major drawbacks of RADFETs is the inability to provide the information on the dose rate in real-time using the conventional absorbed dose measurement technique. The real-time monitoring of dose rate and absorbed dose can be achieved with the current mode dosimeters such as PN and PIN diodes/photodiodes, but these dosimeters have some limitations as absorbed dose meters and hence they are often not a suitable replacement for RADFETs. In that sense, this paper investigates the possibility of using the RADFET as a real-time dose rate meter so that it could be applied for simultaneous online measurement of the dose rate and absorbed dose. A RADFET sample, manufactured by Tyndall National Institute, Cork, Ireland, was tested as a dose rate meter under gamma irradiation from a Co-60 source. The RADFET was configured as a PN junction, such that the drain, gate and source terminals were grounded, while the radiation-induced current was measured at the bulk terminal, whereby the bulk was successively biased with 0 , 10 , 20  and 30 V. In zero-bias mode the radiation-induced current was unstable, but in the biased mode the current response was stable for the investigated dose rates from 0.65  to 32.1 Gy h −1 and up to the total absorbed dose of 25 Gy. The current increased with the dose rate in accordance with the power law, whereas the sensitivity of the current read-out was linear with respect to the applied bias voltage. Comparison with previously analyzed PIN photodiodes has shown that the investigated RADFET is competitive with PIN photodiodes as a gamma radiation dose rate meter and therefore has the potential to be employed for the real-time monitoring of the dose rate and absorbed dose. (paper)

Evaluation of Real-time Measurement Liver Tumor's Movement and SynchronyTM System's Accuracy of Radiosurgery using a Robot CyberKnife

Energy Technology Data Exchange (ETDEWEB)

Kim, Gha Jung; Shim, Su Jung; Kim, Jeong Ho; Min, Chul Kee; Chung, Weon Kuu [Konyang University College of Medicine, Daejeon (Korea, Republic of)

2008-12-15

This study aimed to quantitatively measure the movement of tumors in real-time and evaluate the treatment accuracy, during the treatment of a liver tumor patient, who underwent radiosurgery with a Synchrony Respiratory motion tracking system of a robot CyberKnife. Materials and Methods: The study subjects included 24 liver tumor patients who underwent CyberKnife treatment, which included 64 times of treatment with the Synchrony Respiratory motion tracking system (SynchronyTM). The treatment involved inserting 4 to 6 acupuncture needles into the vicinity of the liver tumor in all the patients using ultrasonography as a guide. A treatment plan was set up using the CT images for treatment planning uses. The position of the acupuncture needle was identified for every treatment time by Digitally Reconstructed Radiography (DRR) prepared at the time of treatment planning and X-ray images photographed in real-time. Subsequent results were stored through a Motion Tracking System (MTS) using the Mtsmain.log treatment file. In this way, movement of the tumor was measured. Besides, the accuracy of radiosurgery using CyberKnife was evaluated by the correlation errors between the real-time positions of the acupuncture needles and the predicted coordinates. Results: The maximum and the average translational movement of the liver tumor were measured 23.5 mm and 13.9{+-}5.5 mm, respectively from the superior to the inferior direction, 3.9 mm and 1.9{+-}0.9 mm, respectively from left to right, and 8.3 mm and 4.9{+-}1.9 mm, respectively from the anterior to the posterior direction. The maximum and the average rotational movement of the liver tumor were measured to be 3.3o and 2.6{+-}1.3o, respectively for X (Left-Right) axis rotation, 4.8o and 2.3{+-}1.0o, respectively for Y (Cranio-Caudal) axis rotation, 3.9o and 2.8{+-}1.1o, respectively for Z (Anterior-Posterior) axis rotation. In addition, the average correlation error, which represents the treatment's accuracy was 1

A two-stage method of quantitative flood risk analysis for reservoir real-time operation using ensemble-based hydrologic forecasts

Science.gov (United States)

Liu, P.

2013-12-01

Quantitative analysis of the risk for reservoir real-time operation is a hard task owing to the difficulty of accurate description of inflow uncertainties. The ensemble-based hydrologic forecasts directly depict the inflows not only the marginal distributions but also their persistence via scenarios. This motivates us to analyze the reservoir real-time operating risk with ensemble-based hydrologic forecasts as inputs. A method is developed by using the forecast horizon point to divide the future time into two stages, the forecast lead-time and the unpredicted time. The risk within the forecast lead-time is computed based on counting the failure number of forecast scenarios, and the risk in the unpredicted time is estimated using reservoir routing with the design floods and the reservoir water levels of forecast horizon point. As a result, a two-stage risk analysis method is set up to quantify the entire flood risks by defining the ratio of the number of scenarios that excessive the critical value to the total number of scenarios. The China's Three Gorges Reservoir (TGR) is selected as a case study, where the parameter and precipitation uncertainties are implemented to produce ensemble-based hydrologic forecasts. The Bayesian inference, Markov Chain Monte Carlo, is used to account for the parameter uncertainty. Two reservoir operation schemes, the real operated and scenario optimization, are evaluated for the flood risks and hydropower profits analysis. With the 2010 flood, it is found that the improvement of the hydrologic forecast accuracy is unnecessary to decrease the reservoir real-time operation risk, and most risks are from the forecast lead-time. It is therefore valuable to decrease the avarice of ensemble-based hydrologic forecasts with less bias for a reservoir operational purpose.

A two-step real-time PCR assay for quantitation and genotyping of human parvovirus 4.

Science.gov (United States)

Väisänen, E; Lahtinen, A; Eis-Hübinger, A M; Lappalainen, M; Hedman, K; Söderlund-Venermo, M

2014-01-01

Human parvovirus 4 (PARV4) of the family Parvoviridae was discovered in a plasma sample of a patient with an undiagnosed acute infection in 2005. Currently, three PARV4 genotypes have been identified, however, with an unknown clinical significance. Interestingly, these genotypes seem to differ in epidemiology. In Northern Europe, USA and Asia, genotypes 1 and 2 have been found to occur mainly in persons with a history of injecting drug use or other parenteral exposure. In contrast, genotype 3 appears to be endemic in sub-Saharan Africa, where it infects children and adults without such risk behaviour. In this study, a novel straightforward and cost-efficient molecular assay for both quantitation and genotyping of PARV4 DNA was developed. The two-step method first applies a single-probe pan-PARV4 qPCR for screening and quantitation of this relatively rare virus, and subsequently, only the positive samples undergo a real-time PCR-based multi-probe genotyping. The new qPCR-GT method is highly sensitive and specific regardless of the genotype, and thus being suitable for studying the clinical impact and occurrence of the different PARV4 genotypes. Copyright © 2013 Elsevier B.V. All rights reserved.

Real-time tricolor phase measuring profilometry based on CCD sensitivity calibration

Science.gov (United States)

Zhu, Lin; Cao, Yiping; He, Dawu; Chen, Cheng

2017-02-01

A real-time tricolor phase measuring profilometry (RTPMP) based on charge coupled device (CCD) sensitivity calibration is proposed. Only one colour fringe pattern whose red (R), green (G) and blue (B) components are, respectively, coded as three sinusoidal phase-shifting gratings with an equivalent shifting phase of 2π/3 is needed and sent to an appointed flash memory on a specialized digital light projector (SDLP). A specialized time-division multiplexing timing sequence actively controls the SDLP to project the fringe patterns in R, G and B channels sequentially onto the measured object in one over seventy-two of a second and meanwhile actively controls a high frame rate monochrome CCD camera to capture the corresponding deformed patterns synchronously with the SDLP. So the sufficient information for reconstructing the three-dimensional (3D) shape in one over twenty-four of a second is obtained. Due to the different spectral sensitivity of the CCD camera to RGB lights, the captured deformed patterns from R, G and B channels cannot share the same peak and valley, which will lead to lower accuracy or even failing to reconstruct the 3D shape. So a deformed pattern amending method based on CCD sensitivity calibration is developed to guarantee the accurate 3D reconstruction. The experimental results verify the feasibility of the proposed RTPMP method. The proposed RTPMP method can obtain the 3D shape at over the video frame rate of 24 frames per second, avoid the colour crosstalk completely and be effective for measuring real-time changing object.

Using Indirect Turbulence Measurements for Real-Time Parameter Estimation in Turbulent Air

Science.gov (United States)

Martos, Borja; Morelli, Eugene A.

2012-01-01

The use of indirect turbulence measurements for real-time estimation of parameters in a linear longitudinal dynamics model in atmospheric turbulence was studied. It is shown that measuring the atmospheric turbulence makes it possible to treat the turbulence as a measured explanatory variable in the parameter estimation problem. Commercial off-the-shelf sensors were researched and evaluated, then compared to air data booms. Sources of colored noise in the explanatory variables resulting from typical turbulence measurement techniques were identified and studied. A major source of colored noise in the explanatory variables was identified as frequency dependent upwash and time delay. The resulting upwash and time delay corrections were analyzed and compared to previous time shift dynamic modeling research. Simulation data as well as flight test data in atmospheric turbulence were used to verify the time delay behavior. Recommendations are given for follow on flight research and instrumentation.

Quantitative Real-time PCR detection of putrescine-producing Gram-negative bacteria

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Kristýna Maršálková

2017-01-01

Full Text Available Biogenic amines are indispensable components of living cells; nevertheless these compounds could be toxic for human health in higher concentrations. Putrescine is supposed to be the major biogenic amine associated with microbial food spoilage. Development of reliable, fast and culture-independent molecular methods to detect bacteria producing biogenic amines deserves the attention, especially of the food industry in purpose to protect health. The objective of this study was to verify the newly designed primer sets for detection of two inducible genes adiA and speF together in Salmonella enterica and Escherichia coli genome by Real-time PCR. These forenamed genes encode enzymes in the metabolic pathway which leads to production of putrescine in Gram-negative bacteria. Moreover, relative expression of these genes was studied in E. coli CCM 3954 strain using Real-time PCR. In this study, sets of new primers for the detection two inducible genes (speF and adiA in Salmonella enterica and E. coli by Real-time PCR were designed and tested. Amplification efficiency of a Real-time PCR was calculated from the slope of the standard curves (adiA, speF, gapA. An efficiency in a range from 95 to 105 % for all tested reactions was achieved. The gene expression (R of adiA and speF genes in E. coli was varied depending on culture conditions. The highest gene expression of adiA and speF was observed at 6, 24 and 36 h (RadiA ~ 3, 5, 9; RspeF ~11, 10, 9; respectively after initiation of growth of this bacteria in nutrient broth medium enchired with amino acids. The results show that these primers could be used for relative quantification analysis of E. coli.

Shelf Life of Food Products: From Open Labeling to Real-Time Measurements.

Science.gov (United States)

Corradini, Maria G

2018-03-25

The labels currently used on food and beverage products only provide consumers with a rough guide to their expected shelf lives because they assume that a product only experiences a limited range of predefined handling and storage conditions. These static labels do not take into consideration conditions that might shorten a product's shelf life (such as temperature abuse), which can lead to problems associated with food safety and waste. Advances in shelf-life estimation have the potential to improve the safety, reliability, and sustainability of the food supply. Selection of appropriate kinetic models and data-analysis techniques is essential to predict shelf life, to account for variability in environmental conditions, and to allow real-time monitoring. Novel analytical tools to determine safety and quality attributes in situ coupled with modern tracking technologies and appropriate predictive tools have the potential to provide accurate estimations of the remaining shelf life of a food product in real time. This review summarizes the necessary steps to attain a transition from open labeling to real-time shelf-life measurements.

In situ real-time measurement of physical characteristics of airborne bacterial particles

Science.gov (United States)

Jung, Jae Hee; Lee, Jung Eun

2013-12-01

Bioaerosols, including aerosolized bacteria, viruses, and fungi, are associated with public health and environmental problems. One promising control method to reduce the harmful effects of bioaerosols is thermal inactivation via a continuous-flow high-temperature short-time (HTST) system. However, variations in bioaerosol physical characteristics - for example, the particle size and shape - during the continuous-flow inactivation process can change the transport properties in the air, which can affect particle deposition in the human respiratory system or the filtration efficiency of ventilation systems. Real-time particle monitoring techniques are a desirable alternative to the time-consuming process of microscopic analysis that is conventionally used in sampling and particle characterization. Here, we report in situ real-time optical scattering measurements of the physical characteristics of airborne bacteria particles following an HTST process in a continuous-flow system. Our results demonstrate that the aerodynamic diameter of bacterial aerosols decreases when exposed to a high-temperature environment, and that the shape of the bacterial cells is significantly altered. These variations in physical characteristics using optical scattering measurements were found to be in agreement with the results of scanning electron microscopy analysis.

A triplex quantitative real-time PCR assay for differential detection of human adenovirus serotypes 2, 3 and 7.

Science.gov (United States)

Qiu, Fang-Zhou; Shen, Xin-Xin; Zhao, Meng-Chuan; Zhao, Li; Duan, Su-Xia; Chen, Chen; Qi, Ju-Ju; Li, Gui-Xia; Wang, Le; Feng, Zhi-Shan; Ma, Xue-Jun

2018-05-02

Human adenovirus (HAdV) serotypes 2, 3 and 7 are more prevalent than other serotypes and have been associated with severe pneumonia in pediatric children. Molecular typing of HAdV is not routinely performed in clinical diagnostic laboratories as it is time-consuming and labor-intensive. In the present study, we developed a triplex quantitative real-time PCR assay (tq-PCR) in a single closed tube for differential detection and quantitative analysis of HAdV serotypes 2, 3 and 7. The sensitivity, specificity, reproducibility and clinical performance of tq-PCR were evaluated. The analytical sensitivity of the tq-PCR was 100 copies/reaction for each of HAdV serotypes 2, 3 and 7, and no cross-reaction with other common respiratory viruses or HAdV serotypes 1,4,5,6,31,55 and 57 was observed. The coefficients of variation (CV) of intra-assay and inter-assay were between 0.6% to 3.6%. Of 138 previously-defined HAdV-positive nasopharyngeal aspirates samples tested, the detection agreement between tq-PCR and nested PCR was 96.38% (133/138). The proposed tq-PCR assay is a sensitive, specific and reproducible method and has the potential for clinical use in the rapid and differential detection and quantitation of HAdV serotypes 2, 3 and 7.

Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR.

Directory of Open Access Journals (Sweden)

Yogita Maheshwari

Full Text Available Droplet digital polymerase chain reaction (ddPCR is a method for performing digital PCR that is based on water-oil emulsion droplet technology. It is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. This study evaluated the applicability of ddPCR as a quantitative detection tool for the Spiroplasma citri, causal agent of citrus stubborn disease (CSD in citrus. Two sets of primers, SP1, based on the spiral in housekeeping gene, and a multicopy prophage gene, SpV1 ORF1, were used to evaluate ddPCR in comparison with real time (quantitative PCR (qPCR for S. citri detection in citrus tissues. Standard curve analyses on tenfold dilution series showed that both ddPCR and qPCR exhibited good linearity and efficiency. However, ddPCR had a tenfold greater sensitivity than qPCR and accurately quantified up to one copy of spiralin gene. Receiver operating characteristic analysis indicated that the ddPCR methodology was more robust for diagnosis of CSD and the area under the curve was significantly broader compared to qPCR. Field samples were used to validate ddPCR efficacy and demonstrated that it was equal or better than qPCR to detect S. citri infection in fruit columella due to a higher pathogen titer. The ddPCR assay detected both the S. citri spiralin and the SpV1 ORF1 targets quantitatively with high precision and accuracy compared to qPCR assay. The ddPCR was highly reproducible and repeatable for both the targets and showed higher resilience to PCR inhibitors in citrus tissue extract for the quantification of S. citri compare to qPCR.

Real time determination of the laser ablated mass by means of electric field-perturbation measurement

Science.gov (United States)

Pacheco, P.; Álvarez, J.; Sarmiento, R.; Bredice, F.; Sánchez-Aké, C.; Villagrán-Muniz, M.; Palleschi, V.

2018-04-01

A Nd:YAG ns-pulsed laser was used to ablate Al, Cd and Zn targets, which were placed between the plates of a planar charged capacitor. The plasma generates a transient redistribution of the electrical charges on the plates that can be measured as a voltage drop across a resistor connected to the ground plate. This signal is proportional to the capacitor applied voltage, the distance between the plates and the total number of ions produced in the ablation process which in turn is related to the laser energy and the ablated mass. After a series of pulses, the targets were weighed on a thermogravimetric balance to measure the ablated mass. Our results show that the electrical signal measured on the resistor is univocally related to the ablated mass from the target. Therefore, after a proper calibration depending on the material and the experimental geometry, the electrical signal can be used for real time quantitative measurement of the ablated mass in pulsed laser generated plasma experiments. The experiments were repeated on an aluminum target, with and without the presence of the external electric field in order to determine the possible influence of the applied electric field on the ablated mass.

Detection, quantitation and identification of enteroviruses from surface waters and sponge tissue from the Florida Keys using real-time RT-PCR

Science.gov (United States)

Donaldson, K.A.; Griffin, Dale W.; Paul, J.H.

2002-01-01

A method was developed for the quantitative detection of pathogenic human enteroviruses from surface waters in the Florida Keys using Taqman (R) one-step Reverse transcription (RT)-PCR with the Model 7700 ABI Prism (R) Sequence Detection System. Viruses were directly extracted from unconcentrated grab samples of seawater, from seawater concentrated by vortex flow filtration using a 100kD filter and from sponge tissue. Total RNA was extracted from the samples, purified and concentrated using spin-column chromatography. A 192-196 base pair portion of the 5??? untranscribed region was amplified from these extracts. Enterovirus concentrations were estimated using real-time RT-PCR technology. Nine of 15 sample sites or 60% were positive for the presence of pathogenic human enteroviruses. Considering only near-shore sites, 69% were positive with viral concentrations ranging from 9.3viruses/ml to 83viruses/g of sponge tissue (uncorrected for extraction efficiency). Certain amplicons were selected for cloning and sequencing for identification. Three strains of waterborne enteroviruses were identified as Coxsackievirus A9, Coxsackievirus A16, and Poliovirus Sabin type 1. Time and cost efficiency of this one-step real-time RT-PCR methodology makes this an ideal technique to detect, quantitate and identify pathogenic enteroviruses in recreational waters. Copyright ?? 2002 Elsevier Science Ltd.

Development and validation of a real-time quantitative PCR assay to detect Xanthomonas axonopodis pv. allii from onion seed.

Science.gov (United States)

Robène, Isabelle; Perret, Marion; Jouen, Emmanuel; Escalon, Aline; Maillot, Marie-Véronique; Chabirand, Aude; Moreau, Aurélie; Laurent, Annie; Chiroleu, Frédéric; Pruvost, Olivier

2015-07-01

Bacterial blight of onion is an emerging disease threatening world onion production. The causal agent Xanthomonas axonopodis pv. allii is seed transmitted and a reliable and sensitive tool is needed to monitor seed exchanges. A triplex quantitative real-time PCR assay was developed targeting two X. axonopodis pv. allii-specific markers and an internal control chosen in 5.8S rRNA gene from Alliaceae. Amplification of at least one marker indicates the presence of the bacterium in seed extracts. This real-time PCR assay detected all the 79 X. axonopodis pv. allii strains tested and excluded 85.2% of the 135 non-target strains and particularly all 39 saprophytic and pathogenic bacteria associated with onion. Cross-reactions were mainly obtained for strains assigned to nine phylogenetically related X. axonopodis pathovars. The cycle cut-off was estimated statistically at 36.3 considering a risk of false positive of 1%. The limit of detection obtained in at least 95% of the time (LOD 95%) was 5×10(3) CFU/g (colony forming unit/g). The sensitivity threshold was found to be 1 infected seed in 32,790 seeds. This real-time PCR assay should be useful for preventing the long-distance spread of X. axonopodis pv. allii via contaminated seed lots and determining the epidemiology of the bacterium. Copyright © 2015 Elsevier B.V. All rights reserved.

Simultaneous quantitative assessment of circulating cell-free mitochondrial and nuclear DNA by multiplex real-time PCR

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Peng Xia

2009-01-01

Full Text Available Quantification of circulating nucleic acids in plasma and serum could be used as a non-invasive diagnostic tool for monitoring a wide variety of diseases and conditions. We describe here a rapid, simple and accurate multiplex real-time PCR method for direct synchronized analysis of circulating cell-free (ccf mitochondrial (mtDNA and nuclear (nDNA DNA in plasma and serum samples. The method is based on one-step multiplex real-time PCR using a FAM-labeled MGB probe and primers to amplify the mtDNA sequence of the ATP 8 gene, and a VIC-labeled MGB probe and primers to amplify the nDNA sequence of the glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH gene, in plasma and serum samples simultaneously. The efficiencies of the multiplex assays were measured in serial dilutions. Based on the simulation of the PCR reaction kinetics, the relative quantities of ccf mtDNA were calculated using a very simple equation. Using our optimised real-time PCR conditions, close to 100% efficiency was obtained from the two assays. The two assays performed in the dilution series showed very good and reproducible correlation to each other. This optimised multiplex real-time PCR protocol can be widely used for synchronized quantification of mtDNA and nDNA in different samples, with a very high rate of efficiency.

Development of quantitative real-time PCR for detection and enumeration of Enterobacteriaceae.

Science.gov (United States)

Takahashi, Hajime; Saito, Rumi; Miya, Satoko; Tanaka, Yuichiro; Miyamura, Natsumi; Kuda, Takashi; Kimura, Bon

2017-04-04

The family Enterobacteriaceae, members of which are widely distributed in the environment, includes many important human pathogens. In this study, a rapid real-time PCR method targeting rplP, coding for L16 protein, a component of the ribosome large subunit, was developed for enumerating Enterobacteriaceae strains, and its efficiency was evaluated using naturally contaminated food products. The rplP-targeted real-time PCR amplified Enterobacteriaceae species with Ct values of 14.0-22.8, whereas the Ct values for non-Enterobacteriaceae species were >30, indicating the specificity of this method for the Enterobacteriaceae. Using a calibration curve of Ct=-3.025 (log CFU/g)+37.35, which was calculated from individual plots of the cell numbers in different concentrations of 5 Enterobacteriaceae species, the rplP-targeted real-time PCR was applied to 51 food samples. A Enterobacteriaceae species in foods rapidly and accurately, and therefore, it can be used for the microbiological risk analysis of foods. Copyright © 2017 Elsevier B.V. All rights reserved.

Real-time measurement and monitoring of absorbed dose for electron beams

Science.gov (United States)

Korenev, Sergey; Korenev, Ivan; Rumega, Stanislav; Grossman, Leon

2004-09-01

The real-time method and system for measurement and monitoring of absorbed dose for industrial and research electron accelerators is considered in the report. The system was created on the basis of beam parameters method. The main concept of this method consists in the measurement of dissipated kinetic energy of electrons in the irradiated product, determination of number of electrons and mass of irradiated product in the same cell by following calculation of absorbed dose in the cell. The manual and automation systems for dose measurements are described. The systems are acceptable for all types of electron accelerators.

Real-time measurement and monitoring of absorbed dose for electron beams

Energy Technology Data Exchange (ETDEWEB)

Korenev, Sergey E-mail: sergey_korenev@steris.com; Korenev, Ivan; Rumega, Stanislav; Grossman, Leon

2004-10-01

The real-time method and system for measurement and monitoring of absorbed dose for industrial and research electron accelerators is considered in the report. The system was created on the basis of beam parameters method. The main concept of this method consists in the measurement of dissipated kinetic energy of electrons in the irradiated product, determination of number of electrons and mass of irradiated product in the same cell by following calculation of absorbed dose in the cell. The manual and automation systems for dose measurements are described. The systems are acceptable for all types of electron accelerators.

Real-time measurement and monitoring of absorbed dose for electron beams

International Nuclear Information System (INIS)

Korenev, Sergey; Korenev, Ivan; Rumega, Stanislav; Grossman, Leon

2004-01-01

The real-time method and system for measurement and monitoring of absorbed dose for industrial and research electron accelerators is considered in the report. The system was created on the basis of beam parameters method. The main concept of this method consists in the measurement of dissipated kinetic energy of electrons in the irradiated product, determination of number of electrons and mass of irradiated product in the same cell by following calculation of absorbed dose in the cell. The manual and automation systems for dose measurements are described. The systems are acceptable for all types of electron accelerators

Comparison between culture and a multiplex quantitative real-time polymerase chain reaction assay detecting Ureaplasma urealyticum and U. parvum.

Science.gov (United States)

Frølund, Maria; Björnelius, Eva; Lidbrink, Peter; Ahrens, Peter; Jensen, Jørgen Skov

2014-01-01

A novel multiplex quantitative real-time polymerase chain reaction (qPCR) for simultaneous detection of U. urealyticum and U. parvum was developed and compared with quantitative culture in Shepard's 10 C medium for ureaplasmas in urethral swabs from 129 men and 66 women, and cervical swabs from 61 women. Using culture as the gold standard, the sensitivity of the qPCR was 96% and 95% for female urethral and cervical swabs, respectively. In male urethral swabs the sensitivity was 89%. The corresponding specificities were 100%, 87% and 99%. The qPCR showed a linear increasing DNA copy number with increasing colour-changing units. Although slightly less sensitive than culture, this multiplex qPCR assay detecting U. urealyticum and U. parvum constitutes a simple and fast alternative to the traditional methods for identification of ureaplasmas and allows simultaneous species differentiation and quantitation in clinical samples. Furthermore, specimens overgrown by other bacteria using the culture method can be evaluated in the qPCR.

Comparison between culture and a multiplex quantitative real-time polymerase chain reaction assay detecting Ureaplasma urealyticum and U. parvum.

Directory of Open Access Journals (Sweden)

Maria Frølund

Full Text Available A novel multiplex quantitative real-time polymerase chain reaction (qPCR for simultaneous detection of U. urealyticum and U. parvum was developed and compared with quantitative culture in Shepard's 10 C medium for ureaplasmas in urethral swabs from 129 men and 66 women, and cervical swabs from 61 women. Using culture as the gold standard, the sensitivity of the qPCR was 96% and 95% for female urethral and cervical swabs, respectively. In male urethral swabs the sensitivity was 89%. The corresponding specificities were 100%, 87% and 99%. The qPCR showed a linear increasing DNA copy number with increasing colour-changing units. Although slightly less sensitive than culture, this multiplex qPCR assay detecting U. urealyticum and U. parvum constitutes a simple and fast alternative to the traditional methods for identification of ureaplasmas and allows simultaneous species differentiation and quantitation in clinical samples. Furthermore, specimens overgrown by other bacteria using the culture method can be evaluated in the qPCR.

An integrated real-time diagnostic concept using expert systems, qualitative reasoning and quantitative analysis

International Nuclear Information System (INIS)

Edwards, R.M.; Lee, K.Y.; Kumara, S.; Levine, S.H.

1989-01-01

An approach for an integrated real-time diagnostic system is being developed for inclusion as an integral part of a power plant automatic control system. In order to participate in control decisions and automatic closed loop operation, the diagnostic system must operate in real-time. Thus far, an expert system with real-time capabilities has been developed and installed on a subsystem at the Experimental Breeder Reactor (EBR-II) in Idaho, USA. Real-time simulation testing of advanced power plant concepts at the Pennsylvania State University has been developed and was used to support the expert system development and installation at EBR-II. Recently, the US National Science Foundation (NSF) and the US Department of Energy (DOE) have funded a Penn State research program to further enhance application of real-time diagnostic systems by pursuing implementation in a distributed power plant computer system including microprocessor based controllers. This paper summarizes past, current, planned, and possible future approaches to power plant diagnostic systems research at Penn State. 34 refs., 9 figs

A Real-Time Measurement System for Long-Life Flood Monitoring and Warning Applications

Directory of Open Access Journals (Sweden)

Antonio Skarmeta Gómez

2012-03-01

Full Text Available A flood warning system incorporates telemetered rainfall and flow/water level data measured at various locations in the catchment area. Real-time accurate data collection is required for this use, and sensor networks improve the system capabilities. However, existing sensor nodes struggle to satisfy the hydrological requirements in terms of autonomy, sensor hardware compatibility, reliability and long-range communication. We describe the design and development of a real-time measurement system for flood monitoring, and its deployment in a flash-flood prone 650 km2 semiarid watershed in Southern Spain. A developed low-power and long-range communication device, so-called DatalogV1, provides automatic data gathering and reliable transmission. DatalogV1 incorporates self-monitoring for adapting measurement schedules for consumption management and to capture events of interest. Two tests are used to assess the success of the development. The results show an autonomous and robust monitoring system for long-term collection of water level data inmany sparse locations during flood events.

Detection and quantification of Renibacterium salmoninarum DNA in salmonid tissues by real-time quantitative polymerase chain reaction analysis

Science.gov (United States)

Chase, D.M.; Elliott, D.G.; Pascho, R.J.

2006-01-01

Renibacterium salmoninarum is an important salmonid pathogen that is difficult to culture. We developed and assessed a real-time, quantitative, polymerase chain reaction (qPCR) assay for the detection and enumeration of R. salmoninarum. The qPCR is based on TaqMan technology and amplifies a 69-base pair (bp) region of the gene encoding the major soluble antigen (MSA) of R. salmoninarum. The qPCR assay consistently detected as few as 5 R. salmoninarum cells per reaction in kidney tissue. The specificity of the qPCR was confirmed by testing the DNA extracts from a panel of microorganisms that were either common fish pathogens or reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA). Kidney samples from 38 juvenile Chinook salmon (Oncorhynchus tshawytscha) in a naturally infected population were examined by real-time qPCR, a nested PCR, and ELISA, and prevalences of R. salmoninarum detected were 71, 66, and 71%, respectively. The qPCR should be a valuable tool for evaluating the R. salmoninarum infection status of salmonids.

Evaluation of Reference Genes for Real-Time Quantitative PCR Analysis in Larvae of Spodoptera litura Exposed to Azadirachtin Stress Conditions

OpenAIRE

Benshui Shu; Jingjing Zhang; Gaofeng Cui; Ranran Sun; Veeran Sethuraman; Xin Yi; Guohua Zhong

2018-01-01

Azadirachtin is an efficient and broad-spectrum botanical insecticide against more than 150 kinds of agricultural pests with the effects of mortality, antifeedant and growth regulation. Real-time quantitative polymerase chain reaction (RT-qPCR) could be one of the powerful tools to analyze the gene expression level and investigate the mechanism of azadirachtin at transcriptional level, however, the ideal reference genes are needed to normalize the expression profiling of target genes. In this...

Sequential processing of quantitative phase images for the study of cell behaviour in real-time digital holographic microscopy.

Science.gov (United States)

Zikmund, T; Kvasnica, L; Týč, M; Křížová, A; Colláková, J; Chmelík, R

2014-11-01

Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The study of the cell is based on extraction of the dynamic data on cell behaviour from the time-lapse sequence of the phase images. However, the phase images are affected by the phase aberrations that make the analysis particularly difficult. This is because the phase deformation is prone to change during long-term experiments. Here, we present a novel algorithm for sequential processing of living cells phase images in a time-lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least-squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. All these procedures are performed automatically and applied immediately after obtaining every single phase image. This property of the algorithm is important for real-time cell quantitative phase imaging and instantaneous control of the course of the experiment by playback of the recorded sequence up to actual time. Such operator's intervention is a forerunner of process automation derived from image analysis. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off-axis holographic microscope. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Real-time heart rate measurement for multi-people using compressive tracking

Science.gov (United States)

Liu, Lingling; Zhao, Yuejin; Liu, Ming; Kong, Lingqin; Dong, Liquan; Ma, Feilong; Pang, Zongguang; Cai, Zhi; Zhang, Yachu; Hua, Peng; Yuan, Ruifeng

2017-09-01

The rise of aging population has created a demand for inexpensive, unobtrusive, automated health care solutions. Image PhotoPlethysmoGraphy(IPPG) aids in the development of these solutions by allowing for the extraction of physiological signals from video data. However, the main deficiencies of the recent IPPG methods are non-automated, non-real-time and susceptible to motion artifacts(MA). In this paper, a real-time heart rate(HR) detection method for multiple subjects simultaneously was proposed and realized using the open computer vision(openCV) library, which consists of getting multiple subjects' facial video automatically through a Webcam, detecting the region of interest (ROI) in the video, reducing the false detection rate by our improved Adaboost algorithm, reducing the MA by our improved compress tracking(CT) algorithm, wavelet noise-suppression algorithm for denoising and multi-threads for higher detection speed. For comparison, HR was measured simultaneously using a medical pulse oximetry device for every subject during all sessions. Experimental results on a data set of 30 subjects show that the max average absolute error of heart rate estimation is less than 8 beats per minute (BPM), and the processing speed of every frame has almost reached real-time: the experiments with video recordings of ten subjects under the condition of the pixel resolution of 600× 800 pixels show that the average HR detection time of 10 subjects was about 17 frames per second (fps).

Development of Real-Time Thickness Measuring System for Insulated Pipeline Using Gamma-ray

International Nuclear Information System (INIS)

Jang, Ji Hoon; Kim, Byung Joo; Cho, Kyung Shik; Kim, Gi Dong

2002-01-01

By this study, on-line real-time radiometric system was developed using a 64 channels linear array of solid state detectors to measure wall thickness of insulated piping system. This system uses an Ir-192 as a gamma ray source and detector is composed of BGO scintillator and photodiode. Ir-192 gamma ray source and linear detector array mounted on a computer controlled robotic crawler. The Ir-192 gamma ray source is located on one side of the piping components and the detector array on the other side. The individual detectors of the detector array measure the intensity of the gamma rays after passing through the walls and the insulation of the piping component under measurement. The output of the detector array is amplified by amplifier and transmitted to the computer through cable. This system collects and analyses the data from the detector array in real-time as the crawler travels over the piping system. The maximum measurable length of pipe is 120cm/min. in the case of 1mm scanning interval

Identification of pyrG Used as an Endogenous Reference Gene in Qualitative and Real-Time Quantitative PCR Detection of Pleurotus ostreatus.

Science.gov (United States)

Zheng, Shi; Shan, Luying; Zhuang, Yongliang; Shang, Ying

2018-03-01

As a well-known edible fungus rich in nutrients, Pleurotus ostreatus has been used as an alternative to expensive wild edible fungi. Specifically, the fact that using P. ostreatus instead of other expensive wild edible fungi has damaged the rights and interests of consumers. Among the existing methods for detection of food adulteration, the amplification of endogenous reference gene is the most accurate method. However, an ideal endogenous reference gene for P. ostreatus has yet to be developed. In this study, a DNA extraction method for P. ostreatus was optimized, and pyrG was selected as a species-specific gene through sequence alignment. This gene was subsequently subjected to qualitative and quantitative Polymerase Chain Reaction (PCR) assays with 3 different P. ostreatus varieties and 7 other species. A low detection limit of 5 pg/μL was obtained by TaqMan quantitative PCR, and no pyrG amplification product was observed in the 7 other species. No allelic variation was detected in P. ostreatus varieties. These experiments confirmed that pyrG was an ideal endogenous reference gene for the qualitative and real-time quantitative PCR detection of P. ostreatus. This method was also suitable for the examination of processed P. ostreatus samples and determination of adulteration in wild mushrooms. The pyrG gene was chosen as an ideal endogenous reference gene for the qualitative and real-time quantitative PCR detection of P. ostreatus, and the detection limit was 5 pg/μL for the quantification. This method is used not only for raw materials but also for processed P. ostreatus products and other processed mushroom foods. © 2018 Institute of Food Technologists®.

Real-time motional Stark effect in jet

International Nuclear Information System (INIS)

Alves, D.; Stephen, A.; Hawkes, N.; Dalley, S.; Goodyear, A.; Felton, R.; Joffrin, E.; Fernandes, H.

2004-01-01

The increasing importance of real-time measurements and control systems in JET experiments, regarding e.g. Internal Transport Barrier (ITB) and q-profile control, has motivated the development of a real-time motional Stark effect (MSE) system. The MSE diagnostic allows the measurement of local magnetic fields in different locations along the neutral beam path providing, therefore, local measurement of the current and q-profiles. Recently in JET, an upgrade of the MSE diagnostic has been implemented, incorporating a totally new system which allows the use of this diagnostic as a real-time control tool as well as an extended data source for off-line analysis. This paper will briefly describe the technical features of the real-time diagnostic with main focus on the system architecture, which consists of a VME crate hosting three PowerPC processor boards and a fast ADC, all connected via Front Panel Data Port (FPDP). The DSP algorithm implements a lockin-amplifier required to demodulate the JET MSE signals. Some applications for the system will be covered such as: feeding the real-time equilibrium reconstruction code (EQUINOX) and allowing the full coverage analysis of the Neutral Beam time window. A brief comparison between the real-time MSE analysis and the off-line analysis will also be presented

A digital approach for real time high-rate high-resolution radiation measurements

International Nuclear Information System (INIS)

Gerardi, G.; Abbene, L.

2014-01-01

Modern spectrometers are currently developed by using digital pulse processing (DPP) systems, showing several advantages over traditional analog electronics. The aim of this work is to present digital strategies, in a time domain, for the development of real time high-rate high-resolution spectrometers. We propose a digital method, based on the single delay line (SDL) shaping technique, able to perform multi-parameter analysis with high performance even at high photon counting rates. A robust pulse shape and height analysis (PSHA), applied on single isolated time windows of the detector output waveforms, is presented. The potentialities of the proposed strategy are highlighted through both theoretical and experimental approaches. To strengthen our approach, the implementation of the method on a real-time system together with some experimental results are presented. X-ray spectra measurements with a semiconductor detector are performed both at low and high photon counting rates (up to 1.1 Mcps)

A digital approach for real time high-rate high-resolution radiation measurements

Energy Technology Data Exchange (ETDEWEB)

Gerardi, G.; Abbene, L., E-mail: leonardo.abbene@unipa.it

2014-12-21

Modern spectrometers are currently developed by using digital pulse processing (DPP) systems, showing several advantages over traditional analog electronics. The aim of this work is to present digital strategies, in a time domain, for the development of real time high-rate high-resolution spectrometers. We propose a digital method, based on the single delay line (SDL) shaping technique, able to perform multi-parameter analysis with high performance even at high photon counting rates. A robust pulse shape and height analysis (PSHA), applied on single isolated time windows of the detector output waveforms, is presented. The potentialities of the proposed strategy are highlighted through both theoretical and experimental approaches. To strengthen our approach, the implementation of the method on a real-time system together with some experimental results are presented. X-ray spectra measurements with a semiconductor detector are performed both at low and high photon counting rates (up to 1.1 Mcps)

Real-time depth measurement for micro-holes drilled by lasers

Science.gov (United States)

Lin, Cheng-Hsiang; Powell, Rock A.; Jiang, Lan; Xiao, Hai; Chen, Shean-Jen; Tsai, Hai-Lung

2010-02-01

An optical system based on the confocal principle has been developed for real-time precision measurements of the depth of micro-holes during the laser drilling process. The capability of the measuring system is theoretically predicted by the Gaussian lens formula and experimentally validated to achieve a sensitivity of 0.5 µm. A nanosecond laser system was used to drill holes, and the hole depths were measured by the proposed measuring system and by the cut-and-polish method. The differences between these two measurements are found to be 5.0% for hole depths on the order of tens of microns and 11.2% for hundreds of microns. The discrepancies are caused mainly by the roughness of the bottom surface of the hole and by the existence of debris in the hole. This system can be easily implemented in a laser workstation for the fabrication of 3D microstructures.

Identification of four squid species by quantitative real-time polymerase chain reaction.

Science.gov (United States)

Ye, Jian; Feng, Junli; Liu, Shasha; Zhang, Yanping; Jiang, Xiaona; Dai, Zhiyuan

2016-02-01

Squids are distributed worldwide, including many species of commercial importance, and they are often made into varieties of flavor foods. The rapid identification methods for squid species especially their processed products, however, have not been well developed. In this study, quantitative real-time PCR (qPCR) systems based on specific primers and TaqMan probes have been established for rapid and accurate identification of four common squid species (Ommastrephes bartramii, Dosidicus gigas, Illex argentinus, Todarodes pacificus) in Chinese domestic market. After analyzing mitochondrial genes reported in GenBank, the mitochondrial cytochrome b (Cytb) gene was selected for O. bartramii detection, cytochrome c oxidase subunit I (COI) gene for D. gigas and T. Pacificus detection, ATPase subunit 6 (ATPase 6) gene for I. Argentinus detection, and 12S ribosomal RNA (12S rDNA) gene for designing Ommastrephidae-specific primers and probe. As a result, all the TaqMan systems are of good performance, and efficiency of each reaction was calculated by making standard curves. This method could detect target species either in single or mixed squid specimen, and it was applied to identify 12 squid processed products successfully. Thus, it would play an important role in fulfilling labeling regulations and squid fishery control. Copyright © 2016 Elsevier Ltd. All rights reserved.

Real-time measurement of dynamic structure for Pd-D system in heavy-water electrolysis cell

International Nuclear Information System (INIS)

Wang Jun; Zeng Xianxin; Yang Jilian; Zhang Baisheng; Ruan Jinghui

1993-01-01

The real-time dynamic structure of Pd-D system in D 2 O electrolysis cell is measured on neutron powder diffractometer in CIAE. Diffraction patterns in 2 θ range of 34 degree-95 degree are obtained under the conditions of electrolysing for 0, 3 and 48 A ·h respectively, and the gradual transition of Pd-D system from α-phase to β-phase is observed. The real-time measurements of β peak of (220) reflection show that intensity of β peak almost reaches the saturation point after electrolysing for 0.65 A · h and increases slowly with further electrolysis afterwards

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.

Infrared Tomography: Data Distribution System for Real-time Mass Flow Rate Measurement

Directory of Open Access Journals (Sweden)

Ruzairi Abdul Rahim

2007-06-01

Full Text Available The system developed in this research has the objective of measuring mass flow rate in an online mode. If a single computer is used as data processing unit, a longer time is needed to produce a measurement result. In the research carried out by previous researcher shows about 11.2 seconds is needed to obtain one mass flow rate result in the offline mode (using offline data. This insufficient real-time result will cause problems in a feedback control process when applying the system on industrial plants. To increase the refreshing rate of the measurement result, an investigation on a data distribution system is performed to replace the existing data processing unit.

Real-time measurement of electron beam weld penetration in uranium by acoustic emission monitoring

International Nuclear Information System (INIS)

Whittaker, J.W.; Murphy, J.L.

1991-07-01

High quality electron beam (EB) welds are required in uranium test articles. Acoustic emission (AE) techniques are under development with the goal of measuring weld penetration in real-time. One technique, based on Average Signal Level (ASL) measurement was used to record weld AE signatures. Characteristic AE signatures were recorded for bead-on-plate (BOP) and butt joint (BJ) welds made under varied welding conditions. AE waveforms were sampled to determine what microscopic AE behavior led to the observed macroscopic signature features. Deformation twinning and weld expulsion are two of the main sources of emission. AE behavior was correlated with weld penetration as measured by standard metallographic techniques. The ASL value was found to increase approximately linearly with weld penetration in BJ welds. These results form the basis for a real-time monitoring technique for weld penetration. 5 refs

Environmental Measurement-While-Drilling system for real-time field screening of contaminants

International Nuclear Information System (INIS)

Lockwood, G.J.; Normann, R.A.; Bishop, L.B.; Floran, R.J.; Williams, C.V.

1995-01-01

Sampling during environmental drilling is essential to fully characterize the spatial distribution and migration of near surface contaminants. However, the analysis of these samples is not only expensive, but can take weeks or months when sent to an off-site laboratory. In contrast, measurement-while-drilling (MWD) screening capability could save money and valuable time by quickly distinguishing between contaminated and uncontaminated areas. Real-time measurements provided by a MVM system would enable on-the-spot decisions to be made regarding sampling strategies, enhance worker safety, and provide the added flexibility of being able to ''steer'' the drill bit in or out hazardous zones. During measurement-while-drilling, down-hole sensors are located behind the drill bit and linked by a rapid data transmission system to a computer at the surface. As drilling proceeds, data are collected on the nature and extent of the subsurface contamination in real-time. The down-hole sensor is a Geiger-Mueller tube (GMT) gamma radiation detector. In addition to the GMT signal, the MWD system monitors these required down-hole voltages and two temperatures associated with the detector assembly. The Gamma Ray Detection System (GRDS) and electronics package are discussed in as well as the results of the field test. Finally, our conclusions and discussion of future work are presented

Estimate of the real-time respiratory simulation system in cyberknife image-guided radiosurgery

International Nuclear Information System (INIS)

Min, Chul Kee; Chung, Weon Kuu; Lee, Suk

2010-01-01

The purpose of this study was to evaluate the target accuracy according to the movement with respiration of an actual patient in a quantitative way by developing a real-time respiratory simulation system (RRSS), including a patient customized 3D moving phantom. The real-time respiratory simulation system (RRSS) consists of two robots in order to implement both the movement of body surfaces and the movement of internal organs caused by respiration. The quantitative evaluation for the 3D movement of the RRSS was performed using a real-time laser displacement sensor for each axis. The average difference in the static movement of the RRSS was about 0.01 ∼ 0.06 mm. Also, in the evaluation of the dynamic movement by producing a formalized sine wave with the phase of four seconds per cycle, the difference between the measured and the calculated values for each cycle length in the robot that was in charge of body surfaces and the robot that was in charge of the movement of internal tumors showed 0.10 ∼ 0.55 seconds, and the correlation coefficients between the calculated and the measured values were 0.998 ∼ 0.999. The differences between the maximum and the minimum amplitudes were 0.01 ∼ 0.06 mm, and the reproducibility was within ±0.5 mm. In the case of the application and non-application of respiration, the target errors were -0.05 ∼ 1.05 mm and -0.13 ∼ 0.74 mm, respectively, and the entire target errors were 1.30 mm and 0.79 mm, respectively. Based on the accuracy in the RRSS system, various respiration patterns of patients can be reproduced in real-time. Also, this system can be used as an optimal tool for applying patient customized accuracy management in image-guided radiosurgery.

Estimate of the real-time respiratory simulation system in cyberknife image-guided radiosurgery

Energy Technology Data Exchange (ETDEWEB)

Min, Chul Kee [Konyang Univ. Hospital, Daejeon (Korea, Republic of); Kyonggi University, Seoul (Korea, Republic of); Chung, Weon Kuu [Konyang Univ. Hospital, Daejeon (Korea, Republic of); Lee, Suk [Korea University, Seoul (Korea, Republic of); and others

2010-01-15

The purpose of this study was to evaluate the target accuracy according to the movement with respiration of an actual patient in a quantitative way by developing a real-time respiratory simulation system (RRSS), including a patient customized 3D moving phantom. The real-time respiratory simulation system (RRSS) consists of two robots in order to implement both the movement of body surfaces and the movement of internal organs caused by respiration. The quantitative evaluation for the 3D movement of the RRSS was performed using a real-time laser displacement sensor for each axis. The average difference in the static movement of the RRSS was about 0.01 {approx} 0.06 mm. Also, in the evaluation of the dynamic movement by producing a formalized sine wave with the phase of four seconds per cycle, the difference between the measured and the calculated values for each cycle length in the robot that was in charge of body surfaces and the robot that was in charge of the movement of internal tumors showed 0.10 {approx} 0.55 seconds, and the correlation coefficients between the calculated and the measured values were 0.998 {approx} 0.999. The differences between the maximum and the minimum amplitudes were 0.01 {approx} 0.06 mm, and the reproducibility was within {+-}0.5 mm. In the case of the application and non-application of respiration, the target errors were -0.05 {approx} 1.05 mm and -0.13 {approx} 0.74 mm, respectively, and the entire target errors were 1.30 mm and 0.79 mm, respectively. Based on the accuracy in the RRSS system, various respiration patterns of patients can be reproduced in real-time. Also, this system can be used as an optimal tool for applying patient customized accuracy management in image-guided radiosurgery.

Real-time safety risk assessment based on a real-time location system for hydropower construction sites.

Science.gov (United States)

Jiang, Hanchen; Lin, Peng; Fan, Qixiang; Qiang, Maoshan

2014-01-01

The concern for workers' safety in construction industry is reflected in many studies focusing on static safety risk identification and assessment. However, studies on real-time safety risk assessment aimed at reducing uncertainty and supporting quick response are rare. A method for real-time safety risk assessment (RTSRA) to implement a dynamic evaluation of worker safety states on construction site has been proposed in this paper. The method provides construction managers who are in charge of safety with more abundant information to reduce the uncertainty of the site. A quantitative calculation formula, integrating the influence of static and dynamic hazards and that of safety supervisors, is established to link the safety risk of workers with the locations of on-site assets. By employing the hidden Markov model (HMM), the RTSRA provides a mechanism for processing location data provided by the real-time location system (RTLS) and analyzing the probability distributions of different states in terms of false positives and negatives. Simulation analysis demonstrated the logic of the proposed method and how it works. Application case shows that the proposed RTSRA is both feasible and effective in managing construction project safety concerns.

Quantitative echocardiographic measures in the assessment of single ventricle function post-Fontan: Incorporation into routine clinical practice.

Science.gov (United States)

Rios, Rodrigo; Ginde, Salil; Saudek, David; Loomba, Rohit S; Stelter, Jessica; Frommelt, Peter

2017-01-01

Quantitative echocardiographic measurements of single ventricular (SV) function have not been incorporated into routine clinical practice. A clinical protocol, which included quantitative measurements of SV deformation (global circumferential and longitudinal strain and strain rate), standard deviation of time to peak systolic strain, myocardial performance index (MPI), dP/dT from an atrioventricular valve regurgitant jet, and superior mesenteric artery resistance index, was instituted for all patients with a history of Fontan procedure undergoing echocardiography. All measures were performed real time during clinically indicated studies and were included in clinical reports. A total of 100 consecutive patients (mean age = 11.95±6.8 years, range 17 months-31.3 years) completed the protocol between September 1, 2014 to April 29, 2015. Deformation measures were completed in 100% of the studies, MPI in 93%, dP/dT in 55%, and superior mesenteric artery Doppler in 82%. The studies were reviewed to assess for efficiency in completing the protocol. The average time for image acquisition was 27.4±8.8 (range 10-62 minutes). The average time to perform deformation measures was 10.8±5.5 minutes (range 5-35 minutes) and time from beginning of imaging to report completion was 53.4±13.7 minutes (range 27-107 minutes). There was excellent inter-observer reliability when deformation indices were blindly repeated. Patients with a single left ventricle had significantly higher circumferential strain and strain rate, longitudinal strain and strain rate, and dP/dT compared to a single right ventricle. There were no differences in quantitative indices of ventricular function between patients 10 years post-Fontan. Advanced quantitative assessment of SV function post-Fontan can be consistently and efficiently performed real time during clinically indicated echocardiograms with excellent reliability. © 2016, Wiley Periodicals, Inc.

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.

Hyperspectral Imaging Using Intracellular Spies: Quantitative Real-Time Measurement of Intracellular Parameters In Vivo during Interaction of the Pathogenic Fungus Aspergillus fumigatus with Human Monocytes.

Directory of Open Access Journals (Sweden)

Sara Mohebbi

Full Text Available Hyperspectral imaging (HSI is a technique based on the combination of classical spectroscopy and conventional digital image processing. It is also well suited for the biological assays and quantitative real-time analysis since it provides spectral and spatial data of samples. The method grants detailed information about a sample by recording the entire spectrum in each pixel of the whole image. We applied HSI to quantify the constituent pH variation in a single infected apoptotic monocyte as a model system. Previously, we showed that the human-pathogenic fungus Aspergillus fumigatus conidia interfere with the acidification of phagolysosomes. Here, we extended this finding to monocytes and gained a more detailed analysis of this process. Our data indicate that melanised A. fumigatus conidia have the ability to interfere with apoptosis in human monocytes as they enable the apoptotic cell to recover from mitochondrial acidification and to continue with the cell cycle. We also showed that this ability of A. fumigatus is dependent on the presence of melanin, since a non-pigmented mutant did not stop the progression of apoptosis and consequently, the cell did not recover from the acidic pH. By conducting the current research based on the HSI, we could measure the intracellular pH in an apoptotic infected human monocyte and show the pattern of pH variation during 35 h of measurements. As a conclusion, we showed the importance of melanin for determining the fate of intracellular pH in a single apoptotic cell.

Charm and J/psi cross section measurements at 13 TeV with real-time calibration

CERN Multimedia

CERN. Geneva

2015-01-01

The LHC's sqrt(s) = 13 TeV proton-proton collisions open a new regime in which the predictions of QCD may be precisely tested via production measurements. LHCb undertook an Early 2015 Measurements campaign to coordinate the operational and analysis activities that are required for rapid completion of such production measurements. The Early Measurements campaign is now bearing fruit with the recent publication of J/psi cross-sections and the imminent publication of charm hadron cross-sections. These are the first results to rely on LHCb's new real-time calibration system, in which the sub-detectors are promptly calibrated and the full event reconstruction of the software High Level Trigger has analysis-quality precision. This seminar will discuss the LHCb real-time calibration system and our recent charm and J/psi production measurements at sqrt(s) = 13 TeV.

The quantification of spermatozoa by real-time quantitative PCR, spectrophotometry, and spermatophore cap size.

Science.gov (United States)

Doyle, Jacqueline M; McCormick, Cory R; DeWoody, J Andrew

2011-01-01

Many animals, such as crustaceans, insects, and salamanders, package their sperm into spermatophores, and the number of spermatozoa contained in a spermatophore is relevant to studies of sexual selection and sperm competition. We used two molecular methods, real-time quantitative polymerase chain reaction (RT-qPCR) and spectrophotometry, to estimate sperm numbers from spermatophores. First, we designed gene-specific primers that produced a single amplicon in four species of ambystomatid salamanders. A standard curve generated from cloned amplicons revealed a strong positive relationship between template DNA quantity and cycle threshold, suggesting that RT-qPCR could be used to quantify sperm in a given sample. We then extracted DNA from multiple Ambystoma maculatum spermatophores, performed RT-qPCR on each sample, and estimated template copy numbers (i.e. sperm number) using the standard curve. Second, we used spectrophotometry to determine the number of sperm per spermatophore by measuring DNA concentration relative to the genome size. We documented a significant positive relationship between the estimates of sperm number based on RT-qPCR and those based on spectrophotometry. When these molecular estimates were compared to spermatophore cap size, which in principle could predict the number of sperm contained in the spermatophore, we also found a significant positive relationship between sperm number and spermatophore cap size. This linear model allows estimates of sperm number strictly from cap size, an approach which could greatly simplify the estimation of sperm number in future studies. These methods may help explain variation in fertilization success where sperm competition is mediated by sperm quantity. © 2010 Blackwell Publishing Ltd.

Real-Time Particle Mass Spectrometry Based on Resonant Micro Strings

DEFF Research Database (Denmark)

Schmid, Silvan; Dohn, Søren; Boisen, Anja

2010-01-01

by measuring the resonant frequency shifts of the first two bending modes. The method has been tested by detecting the mass spectrum of micro particles placed on a micro string. This method enables real-time mass spectrometry necessary for applications such as personal monitoring devices for the assessment......Micro- and nanomechanical resonators are widely being used as mass sensors due to their unprecedented mass sensitivity. We present a simple closed-form expression which allows a fast and quantitative calculation of the position and mass of individual particles placed on a micro or nano string...

Real-time adjustment of ventricular restraint therapy in heart failure.

Science.gov (United States)

Ghanta, Ravi K; Lee, Lawrence S; Umakanthan, Ramanan; Laurence, Rita G; Fox, John A; Bolman, Ralph Morton; Cohn, Lawrence H; Chen, Frederick Y

2008-12-01

Current ventricular restraint devices do not allow for either the measurement or adjustment of ventricular restraint level. Periodic adjustment of restraint level post-device implantation may improve therapeutic efficacy. We evaluated the feasibility of an adjustable quantitative ventricular restraint (QVR) technique utilizing a fluid-filled polyurethane epicardial balloon to measure and adjust restraint level post-implantation guided by physiologic parameters. QVR balloons were implanted in nine ovine with post-infarction dilated heart failure. Restraint level was defined by the maximum restraint pressure applied by the balloon to the epicardium at end-diastole. An access line connected the balloon lumen to a subcutaneous portacath to allow percutaneous access. Restraint level was adjusted while left ventricular (LV) end-diastolic volume (EDV) and cardiac output was assessed with simultaneous transthoracic echocardiography. All nine ovine successfully underwent QVR balloon implantation. Post-implantation, restraint level could be measured percutaneously in real-time and dynamically adjusted by instillation and withdrawal of fluid from the balloon lumen. Using simultaneous echocardiography, restraint level could be adjusted based on LV EDV and cardiac output. After QVR therapy for 21 days, LV EDV decreased from 133+/-15 ml to 113+/-17 ml (p<0.05). QVR permits real-time measurement and physiologic adjustment of ventricular restraint therapy after device implantation.

A study of real-time content marketing : formulating real-time content marketing based on content, search and social media

OpenAIRE

Nguyen, Thi Kim Duyen

2015-01-01

The primary objective of this research is to understand profoundly the new concept of content marketing – real-time content marketing on the aspect of the digital marketing experts. Particularly, the research will focus on the real-time content marketing theories and how to build real-time content marketing strategy based on content, search and social media. It also finds out how marketers measure and keep track of conversion rates of their real-time content marketing plan. Practically, th...

Real-time measurement of dust in the workplace using video exposure monitoring: Farming to pharmaceuticals

International Nuclear Information System (INIS)

Walsh, P T; Forth, A R; Clark, R D R; Dowker, K P; Thorpe, A

2009-01-01

Real-time, photometric, portable dust monitors have been employed for video exposure monitoring (VEM) to measure and highlight dust levels generated by work activities, illustrate dust control techniques, and demonstrate good practice. Two workplaces, presenting different challenges for measurement, were used to illustrate the capabilities of VEM: (a) poultry farming activities and (b) powder transfer operations in a pharmaceutical company. For the poultry farm work, the real-time monitors were calibrated with respect to the respirable and inhalable dust concentrations using cyclone and IOM reference samplers respectively. Different rankings of exposure for typical activities were found on the small farm studied here compared to previous exposure measurements at larger poultry farms: these were mainly attributed to the different scales of operation. Large variations in the ratios of respirable, inhalable and real-time monitor TWA concentrations of poultry farm dust for various activities were found. This has implications for the calibration of light-scattering dust monitors with respect to inhalable dust concentration. In the pharmaceutical application, the effectiveness of a curtain barrier for dust control when dispensing powder in a downflow booth was rapidly demonstrated.

Real-Time, Single-Shot Temporal Measurements of Short Electron Bunches, Terahertz CSR and FEL Radiation

CERN Document Server

Berden, G; Van der Meer, A F G

2005-01-01

Electro-optic detection of the Coulomb field of electron bunches is a promising technique for single-shot measurements of the bunch length and shape in the sub-picosecond time domain. This technique has been applied to the measurement of 50 MeV electron bunches in the FELIX free electron laser, showing the longitudinal profile of single bunches of around 650 fs FWHM [Phys. Rev. Lett. 93, 114802 (2004)]. The method is non-destructive and real-time, and therefore ideal for online monitoring of the longitudinal shape of single electron bunches. At FELIX we have used it for real-time optimization of sub-picosecond electron bunches. Electro-optic detection has also been used to measure the electric field profiles of far-infrared (or terahertz) optical pulses generated by the relativistic electrons. We have characterised the far-infrared output of the free electron laser, and more recently, we have measured the temporal profile of terahertz optical pulses generated at one of the bending magnets.

A method for accurate detection of genomic microdeletions using real-time quantitative PCR

Directory of Open Access Journals (Sweden)

Bassett Anne S

2005-12-01

Full Text Available Abstract Background Quantitative Polymerase Chain Reaction (qPCR is a well-established method for quantifying levels of gene expression, but has not been routinely applied to the detection of constitutional copy number alterations of human genomic DNA. Microdeletions or microduplications of the human genome are associated with a variety of genetic disorders. Although, clinical laboratories routinely use fluorescence in situ hybridization (FISH to identify such cryptic genomic alterations, there remains a significant number of individuals in which constitutional genomic imbalance is suspected, based on clinical parameters, but cannot be readily detected using current cytogenetic techniques. Results In this study, a novel application for real-time qPCR is presented that can be used to reproducibly detect chromosomal microdeletions and microduplications. This approach was applied to DNA from a series of patient samples and controls to validate genomic copy number alteration at cytoband 22q11. The study group comprised 12 patients with clinical symptoms of chromosome 22q11 deletion syndrome (22q11DS, 1 patient trisomic for 22q11 and 4 normal controls. 6 of the patients (group 1 had known hemizygous deletions, as detected by standard diagnostic FISH, whilst the remaining 6 patients (group 2 were classified as 22q11DS negative using the clinical FISH assay. Screening of the patients and controls with a set of 10 real time qPCR primers, spanning the 22q11.2-deleted region and flanking sequence, confirmed the FISH assay results for all patients with 100% concordance. Moreover, this qPCR enabled a refinement of the region of deletion at 22q11. Analysis of DNA from chromosome 22 trisomic sample demonstrated genomic duplication within 22q11. Conclusion In this paper we present a qPCR approach for the detection of chromosomal microdeletions and microduplications. The strategic use of in silico modelling for qPCR primer design to avoid regions of repetitive

Culture-independent identification and quantification of Gallibacterium anatis (G. anatis) by real-time quantitative PCR

DEFF Research Database (Denmark)

Wang, Chong; Robles, Francisco; Ramirez, Saul

2016-01-01

Gallibacterium is a genus within the family Pasteurellaceae characterized by a high level of phenotypic and genetic diversity. No diagnostic method has yet been described, which allows species-specific identification of Gallibacterium anatis. The aim of this study was to develop a real...... published conventional PCR method and culture-based identification, respectively. The detection rates were 97%, 78% and 34% for the current qPCR, the conventional PCR and the culture-based identification method, respectively. The qPCR assay was able to detect the gene gyrB in serial dilutions of 10......-time quantitative PCR (qPCR) method allowing species-specific identification and quantification of G. anatis. A G. anatis specific DNA sequence was identified in the gyrase subunit B gene (gyrB) and used to design a TaqMan probe and corresponding primers. The specificity of the assay was tested on 52 bacterial...

Real-time measurements to characterize dynamics of emulsion interface during simulated intestinal digestion.

Science.gov (United States)

Pan, Yuanjie; Nitin, N

2016-05-01

Efficient delivery of bioactives remains a critical challenge due to their limited bioavailability and solubility. While many encapsulation systems are designed to modulate the digestion and release of bioactives within the human gastrointestinal tract, there is limited understanding of how engineered structures influence the delivery of bioactives. The objective of this study was to develop a real-time quantitative method to measure structural changes in emulsion interface during simulated intestinal digestion and to correlate these changes with the release of free fatty acids (FFAs). Fluorescence resonant energy transfer (FRET) was used for rapid in-situ measurement of the structural changes in emulsion interface during simulated intestinal digestion. By using FRET, changes in the intermolecular spacing between the two different fluorescent probes labeled emulsifier were characterized. Changes in FRET measurements were compared with the release of FFAs. The results showed that bile salts and pancreatic lipase interacted immediately with the emulsion droplets and disrupted the emulsion interface as evidenced by reduction in FRET efficacy compared to the control. Similarly, a significant amount of FFAs was released during digestion. Moreover, addition of a second layer of polymers at emulsion interface decreased the extent of interface disruption by bile salts and pancreatic lipase and impacted the amount or rate of FFA release during digestion. These results were consistent with the lower donor/acceptor ratio of the labeled probes from the FRET result. Overall, this study provides a novel approach to analyze the dynamics of emulsion interface during digestion and their relationship with the release of FFAs. Copyright © 2016 Elsevier B.V. All rights reserved.

Real-Time PCR-Based Quantitation Method for the Genetically Modified Soybean Line GTS 40-3-2.

Science.gov (United States)

Kitta, Kazumi; Takabatake, Reona; Mano, Junichi

2016-01-01

This chapter describes a real-time PCR-based method for quantitation of the relative amount of genetically modified (GM) soybean line GTS 40-3-2 [Roundup Ready(®) soybean (RRS)] contained in a batch. The method targets a taxon-specific soybean gene (lectin gene, Le1) and the specific DNA construct junction region between the Petunia hybrida chloroplast transit peptide sequence and the Agrobacterium 5-enolpyruvylshikimate-3-phosphate synthase gene (epsps) sequence present in GTS 40-3-2. The method employs plasmid pMulSL2 as a reference material in order to quantify the relative amount of GTS 40-3-2 in soybean samples using a conversion factor (Cf) equal to the ratio of the RRS-specific DNA to the taxon-specific DNA in representative genuine GTS 40-3-2 seeds.

Strategy for Extracting DNA from Clay Soil and Detecting a Specific Target Sequence via Selective Enrichment and Real-Time (Quantitative) PCR Amplification ▿

Science.gov (United States)

Yankson, Kweku K.; Steck, Todd R.

2009-01-01

We present a simple strategy for isolating and accurately enumerating target DNA from high-clay-content soils: desorption with buffers, an optional magnetic capture hybridization step, and quantitation via real-time PCR. With the developed technique, μg quantities of DNA were extracted from mg samples of pure kaolinite and a field clay soil. PMID:19633108

Real-time power angle determination of salient-pole synchronous machine based on air gap measurements

Energy Technology Data Exchange (ETDEWEB)

Despalatovic, Marin; Jadric, Martin; Terzic, Bozo [FESB University of Split, Faculty of Electrical Engineering, Mechanical Engineering and Naval Architecture, R. Boskovica bb, 21000 Split (Croatia)

2008-11-15

This paper presents a new method for the real-time power angle determination of the salient-pole synchronous machines. This method is based on the terminal voltage and air gap measurements, which are the common features of the hydroturbine generator monitoring system. The raw signal of the air gap sensor is used to detect the rotor displacement with reference to the fundamental component of the terminal voltage. First, the algorithm developed for the real-time power angle determination is tested using the synthetic data obtained by the standard machine model simulation. Thereafter, the experimental investigation is carried out on the 26 MVA utility generator. The validity of the method is verified by comparing with another method, which is based on a tooth gear mounted on the rotor shaft. The proposed real-time algorithm has an adequate accuracy and needs a very short processing time. For applications that do not require real-time processing, such as the estimation of the synchronous machine parameters, the accuracy is additionally increased by applying an off-line data-processing algorithm. (author)

A novel approach for evaluating the performance of real time quantitative loop-mediated isothermal amplification-based methods.

Science.gov (United States)

Nixon, Gavin J; Svenstrup, Helle F; Donald, Carol E; Carder, Caroline; Stephenson, Judith M; Morris-Jones, Stephen; Huggett, Jim F; Foy, Carole A

2014-12-01

Molecular diagnostic measurements are currently underpinned by the polymerase chain reaction (PCR). There are also a number of alternative nucleic acid amplification technologies, which unlike PCR, work at a single temperature. These 'isothermal' methods, reportedly offer potential advantages over PCR such as simplicity, speed and resistance to inhibitors and could also be used for quantitative molecular analysis. However there are currently limited mechanisms to evaluate their quantitative performance, which would assist assay development and study comparisons. This study uses a sexually transmitted infection diagnostic model in combination with an adapted metric termed isothermal doubling time (IDT), akin to PCR efficiency, to compare quantitative PCR and quantitative loop-mediated isothermal amplification (qLAMP) assays, and to quantify the impact of matrix interference. The performance metric described here facilitates the comparison of qLAMP assays that could assist assay development and validation activities.

Field instruments for real time in-situ crude oil concentration measurements

International Nuclear Information System (INIS)

Fuller, C.B.; Bonner, J.S.; Page, C.A.; Arrambide, G.; Sterling, M.C.Jr.; Ojo, T.O.

2003-01-01

Accidental oil spills, contaminant release during resuspension, storms, and harmful algal blooms are all episodic events that can effect coastal margins. It is important to quantitatively describe water and ecological quality evolution and predict the impact to these areas by such events, but traditional sampling methods miss environmental activity during cyclical events. This paper presents a new sampling approach that involves continuous, real-time in-situ monitoring to provide data for development of comprehensive modeling protocols. It gives spill response coordinators greater assurance in making decisions using the latest visualization tools which are based on a good understanding of the physical processes at work in pulsed events. Five sensors for rapid monitoring of crude oil concentrations in aquatic systems were described. The in-situ and ex-situ sensors can measure plume transport and estimate polycyclic aromatic hydrocarbon exposure concentrations to assess risk of toxicity. A brief description and evaluation of the following 5 sensors was provided: the LISST-100 by Sequoia Instrument, a submersible multi-angle laser scattering instrument; the AU-10 field fluorometer by Turner Designs, an ex-situ single wavelength fluorometer; the Flashlamp by WET Labs Inc., an in-situ single wavelength fluorometer; and, the ECO-FL3 and SAFire by WET Labs Inc., two in-situ multiple wavelength fluorometers. These instruments were used to analyze crude oil emissions of various concentrations. All of the instruments followed a linear response within the tested concentration range. At the lowest concentrations the LISST-100 was not as effective as the fluorometers because of limited particle volume for scatter. For the AU-10 field fluorometer, the highest concentrations tested were above the measurement range of the instrument. 6 refs., 5 figs

Identification of real-time diagnostic measures of visual distraction with an automatic eye-tracking system.

Science.gov (United States)

Zhang, Harry; Smith, Matthew R H; Witt, Gerald J

2006-01-01

This study was conducted to identify eye glance measures that are diagnostic of visual distraction. Visual distraction degrades performance, but real-time diagnostic measures have not been identified. In a driving simulator, 14 participants responded to a lead vehicle braking at -2 or -2.7 m/s2 periodically while reading a varying number of words (6-15 words every 13 s) on peripheral displays (with diagonal eccentricities of 24 degrees, 43 degrees, and 75 degrees). As the number of words and display eccentricity increased, total glance duration and reaction time increased and driving performance suffered. Correlation coefficients between several glance measures and reaction time or performance variables were reliably high, indicating that these glance measures are diagnostic of visual distraction. It is predicted that for every 25% increase in total glance duration, reaction time is increased by 0.39 s and standard deviation of lane position is increased by 0.06 m. Potential applications of this research include assessing visual distraction in real time, delivering advisories to distracted drivers to reorient their attention to driving, and using distraction information to adapt forward collision and lane departure warning systems to enhance system effectiveness.

High-Resolution Light Transmission Spectroscopy of Nanoparticles in Real Time

Science.gov (United States)

Tanner, Carol; Sun, Nan; Deatsch, Alison; Li, Frank; Ruggiero, Steven

2017-04-01

As implemented here, Light Transmission Spectroscopy (LTS) is a high-resolution real-time technique for eliminating spectral noise and systematic effects in wide band spectroscopic measurements of nanoparticles. In this work, we combine LTS with spectral inversion for the purpose of characterizing the size, shape, and number of nanoparticles in solution. The apparatus employs a wide-band multi-wavelength light source and grating spectrometers coupled to CCD detectors. The light source ranges from 210 to 2000 nm, and the wavelength dependent light detection system ranges from 200 to 1100 nm with model the total extinction cross-section, and spectral inversion is employed to obtain quantitative particle size distributions. Discussed are the precision, accuracy, resolution, and sensitivity of our results. The technique is quite versatile and can be applied to spectroscopic investigations where wideband, accurate, low-noise, real-time spectra are desired. University of Notre Dame Office of Research, College of Science, Department of Physics, and USDA.

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.

Quantitative real-time PCR identifies a critical region of deletion on 22q13 related to prognosis in oral cancer

DEFF Research Database (Denmark)

Reis, Patricia P; Rogatto, Silvia R; Kowalski, Luiz P

2002-01-01

Quantitative real time PCR was performed on genomic DNA from 40 primary oral carcinomas and the normal adjacent tissues. The target genes ECGFB, DIA1, BIK, and PDGFB and the microsatellite markers D22S274 and D22S277, mapped on 22q13, were selected according to our previous loss of heterozygosity...... findings in head and neck tumors. Quantitative PCR relies on the comparison of the amount of product generated from a target gene and that generated from a disomic reference gene (GAPDH-housekeeping gene). Reactions have been performed with normal control in triplicates, using the 7700 Sequence Detection.......0018) for patients with DIA1 gene loss. Relative copy number losses detected in these sequences may be related to disease progression and a worse prognosis in patients with oral cancer....

Development of a real-time stability measurement system for boiling water reactors

International Nuclear Information System (INIS)

March-Leuba, J.; King, W.T.

1987-01-01

This paper describes the development of a portable, real time system for boiling water reactor (BWR) stability measurements. The system provides a means for the operator to monitor the reactor stability using existing plant instrumentation and commercially available hardware. The noise component (i.e., perturbations around steady state) of the neutron signal in BWRs has been shown to contain information about reactor stability, and several algorithms have been developed to extract that information. For the present work, the authors have used an algorithm that has been implemented on a portable personal computer. This algorithm uses the autocorrelation function of naturally occurring neutron noise (measured without special plant perturbations) and an autoregressive modeling technique to produce the asymptotic DR. For this real-time implementation, neutron noise data is preconditioned (i.e., filtered and amplified) and sampled at a 5-Hz sampling rate using a commercial data-acquisition system. Approximately every 1.5 min, the current (snapshot) autocorrelation function is computed directly from the data, and the average autocorrelation is updated. The current and average DR estimates are evaluated with the same periodicity and are displayed on the screen along with the autocorrelations and average power spectrum of the neutron noise

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.

Near-real-time materials accountancy in an international perspective or will the real near-real-time materials accountancy please stand up

International Nuclear Information System (INIS)

Lovett, J.E.

1981-01-01

During the 1970's the IAEA gave considerable attention to the question of what its quantitative goals should be. These discussions led, in January 1980, to a set of provisional guidelines which considered both abrupt and protracted diversion possibilities. In a search for an effective means of achieving these goals at large bulk processing facilities, two technological concepts have been put forward. One concept, commonly referred to as near-real-time materials accountancy, is reviewed in this paper. 9 refs

Development of a Capacitive Ice Sensor to Measure Ice Growth in Real Time

Directory of Open Access Journals (Sweden)

Xiang Zhi

2015-03-01

Full Text Available This paper presents the development of the capacitive sensor to measure the growth of ice on a fuel pipe surface in real time. The ice sensor consists of pairs of electrodes to detect the change in capacitance and a thermocouple temperature sensor to examine the ice formation situation. In addition, an environmental chamber was specially designed to control the humidity and temperature to simulate the ice formation conditions. From the humidity, a water film is formed on the ice sensor, which results in an increase in capacitance. Ice nucleation occurs, followed by the rapid formation of frost ice that decreases the capacitance suddenly. The capacitance is saturated. The developed ice sensor explains the ice growth providing information about the icing temperature in real time.

Development of a capacitive ice sensor to measure ice growth in real time.

Science.gov (United States)

Zhi, Xiang; Cho, Hyo Chang; Wang, Bo; Ahn, Cheol Hee; Moon, Hyeong Soon; Go, Jeung Sang

2015-03-19

This paper presents the development of the capacitive sensor to measure the growth of ice on a fuel pipe surface in real time. The ice sensor consists of pairs of electrodes to detect the change in capacitance and a thermocouple temperature sensor to examine the ice formation situation. In addition, an environmental chamber was specially designed to control the humidity and temperature to simulate the ice formation conditions. From the humidity, a water film is formed on the ice sensor, which results in an increase in capacitance. Ice nucleation occurs, followed by the rapid formation of frost ice that decreases the capacitance suddenly. The capacitance is saturated. The developed ice sensor explains the ice growth providing information about the icing temperature in real time.

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.

A novel approach for evaluating the performance of real time quantitative loop-mediated isothermal amplification-based methods

Directory of Open Access Journals (Sweden)

Gavin J. Nixon

2014-12-01

Full Text Available Molecular diagnostic measurements are currently underpinned by the polymerase chain reaction (PCR. There are also a number of alternative nucleic acid amplification technologies, which unlike PCR, work at a single temperature. These ‘isothermal’ methods, reportedly offer potential advantages over PCR such as simplicity, speed and resistance to inhibitors and could also be used for quantitative molecular analysis. However there are currently limited mechanisms to evaluate their quantitative performance, which would assist assay development and study comparisons. This study uses a sexually transmitted infection diagnostic model in combination with an adapted metric termed isothermal doubling time (IDT, akin to PCR efficiency, to compare quantitative PCR and quantitative loop-mediated isothermal amplification (qLAMP assays, and to quantify the impact of matrix interference. The performance metric described here facilitates the comparison of qLAMP assays that could assist assay development and validation activities.

IoT for Real-Time Measurement of High-Throughput Liquid Dispensing in Laboratory Environments.

Science.gov (United States)

Shumate, Justin; Baillargeon, Pierre; Spicer, Timothy P; Scampavia, Louis

2018-04-01

Critical to maintaining quality control in high-throughput screening is the need for constant monitoring of liquid-dispensing fidelity. Traditional methods involve operator intervention with gravimetric analysis to monitor the gross accuracy of full plate dispenses, visual verification of contents, or dedicated weigh stations on screening platforms that introduce potential bottlenecks and increase the plate-processing cycle time. We present a unique solution using open-source hardware, software, and 3D printing to automate dispenser accuracy determination by providing real-time dispense weight measurements via a network-connected precision balance. This system uses an Arduino microcontroller to connect a precision balance to a local network. By integrating the precision balance as an Internet of Things (IoT) device, it gains the ability to provide real-time gravimetric summaries of dispensing, generate timely alerts when problems are detected, and capture historical dispensing data for future analysis. All collected data can then be accessed via a web interface for reviewing alerts and dispensing information in real time or remotely for timely intervention of dispense errors. The development of this system also leveraged 3D printing to rapidly prototype sensor brackets, mounting solutions, and component enclosures.

Comparison of high and low virulence serotypes of Actinobacillus pleuropneumoniae by quantitative real-time PCR

DEFF Research Database (Denmark)

Schou, Kirstine Klitgaard; Angen, Øystein; Boye, Mette

PCR data. Preliminary results showed that in both serotype 2 and serotype 6, the toxin producing gene apxIV was the most highly expressed of the investigated genes. The major difference observed between the two serotypes was that apfA, involved in type IV vili production, was significantly upregulated...... of high virulence while serotype 6 strains are normally found to be less pathogenic. To gain an understanding of the differential virulence of serotype 2 and 6, the expression of a panel of Ap genes during infection of porcine epithelial lung cells (SJPL) were examined by quantitative real-time PCR (q...... to be important for early establishment of the bacteria in the host were examined by qPCR. The genes examined were apfA, coding for a subunit of Type IV pili, kdsB coding for a gene involved in lippopolysacceride biosynthesis, and pgaB which is involved in biofilm formation, all three believed to be important...

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

DEFF Research Database (Denmark)

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

2014-01-01

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

Single-cell real-time imaging of transgene expression upon lipofection

Energy Technology Data Exchange (ETDEWEB)

Fiume, Giuseppe [Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127 Pisa (Italy); Di Rienzo, Carmine [Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127 Pisa (Italy); NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Piazza San Silvestro 12, 56127, Pisa (Italy); Marchetti, Laura [Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127 Pisa (Italy); Pozzi, Daniela; Caracciolo, Giulio [Department of Molecular Medicine, “Sapienza” University of Rome, Viale Regina Elena 291, 00161, Rome (Italy); Cardarelli, Francesco, E-mail: francesco.cardarelli@iit.it [Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127 Pisa (Italy)

2016-05-20

Here we address the process of lipofection by quantifying the expression of a genetically-encoded fluorescent reporter at the single-cell level, and in real-time, by confocal imaging in live cells. The Lipofectamine gold-standard formulation is compared to the alternative promising DC-Chol/DOPE formulation. In both cases, we report that only dividing cells are able to produce a detectable amount of the fluorescent reporter protein. Notably, by measuring fluorescence over time in each pair of daughter cells, we find that Lipofectamine-based transfection statistically yields a remarkably higher degree of “symmetry” in protein expression between daughter cells as compared to DC-Chol/DOPE. A model is envisioned in which the degree of symmetry of protein expression is linked to the number of bioavailable DNA copies within the cell before nuclear breakdown. Reported results open new perspectives for the understanding of the lipofection mechanism and define a new experimental platform for the quantitative comparison of transfection reagents. -- Highlights: •The process of lipofection is followed by quantifying the transgene expression in real time. •The Lipofectamine gold-standard is compared to the promising DC-Chol/DOPE formulation. •We report that only dividing cells are able to produce the fluorescent reporter protein. •The degree of symmetry of protein expression in daughter cells is linked to DNA bioavailability. •A new experimental platform for the quantitative comparison of transfection reagents is proposed.

Single-cell real-time imaging of transgene expression upon lipofection

International Nuclear Information System (INIS)

Fiume, Giuseppe; Di Rienzo, Carmine; Marchetti, Laura; Pozzi, Daniela; Caracciolo, Giulio; Cardarelli, Francesco

2016-01-01

Here we address the process of lipofection by quantifying the expression of a genetically-encoded fluorescent reporter at the single-cell level, and in real-time, by confocal imaging in live cells. The Lipofectamine gold-standard formulation is compared to the alternative promising DC-Chol/DOPE formulation. In both cases, we report that only dividing cells are able to produce a detectable amount of the fluorescent reporter protein. Notably, by measuring fluorescence over time in each pair of daughter cells, we find that Lipofectamine-based transfection statistically yields a remarkably higher degree of “symmetry” in protein expression between daughter cells as compared to DC-Chol/DOPE. A model is envisioned in which the degree of symmetry of protein expression is linked to the number of bioavailable DNA copies within the cell before nuclear breakdown. Reported results open new perspectives for the understanding of the lipofection mechanism and define a new experimental platform for the quantitative comparison of transfection reagents. -- Highlights: •The process of lipofection is followed by quantifying the transgene expression in real time. •The Lipofectamine gold-standard is compared to the promising DC-Chol/DOPE formulation. •We report that only dividing cells are able to produce the fluorescent reporter protein. •The degree of symmetry of protein expression in daughter cells is linked to DNA bioavailability. •A new experimental platform for the quantitative comparison of transfection reagents is proposed.

Real time alpha value measurement with Feynman-α method utilizing time series data acquisition on low enriched uranium system

International Nuclear Information System (INIS)

Tonoike, Kotaro; Yamamoto, Toshihiro; Watanabe, Shoichi; Miyoshi, Yoshinori

2003-01-01

As a part of the development of a subcriticality monitoring system, a system which has a time series data acquisition function of detector signals and a real time evaluation function of alpha value with the Feynman-alpha method was established, with which the kinetic parameter (alpha value) was measured at the STACY heterogeneous core. The Hashimoto's difference filter was implemented in the system, which enables the measurement at a critical condition. The measurement result of the new system agreed with the pulsed neutron method. (author)

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.

Quantitative real-time PCR approaches for microbial community studies in wastewater treatment systems: applications and considerations.

Science.gov (United States)

Kim, Jaai; Lim, Juntaek; Lee, Changsoo

2013-12-01

Quantitative real-time PCR (qPCR) has been widely used in recent environmental microbial ecology studies as a tool for detecting and quantifying microorganisms of interest, which aids in better understandings of the complexity of wastewater microbial communities. Although qPCR can be used to provide more specific and accurate quantification than other molecular techniques, it does have limitations that must be considered when applying it in practice. This article reviews the principle of qPCR quantification and its applications to microbial ecology studies in various wastewater treatment environments. Here we also address several limitations of qPCR-based approaches that can affect the validity of quantification data: template nucleic acid quality, nucleic acid extraction efficiency, specificity of group-specific primers and probes, amplification of nonviable DNA, gene copy number variation, and limited number of sequences in the database. Even with such limitations, qPCR is reportedly among the best methods for quantitatively investigating environmental microbial communities. The application of qPCR is and will continue to be increasingly common in studies of wastewater treatment systems. To obtain reliable analyses, however, the limitations that have often been overlooked must be carefully considered when interpreting the results. Copyright © 2013 Elsevier Inc. All rights reserved.

Optimized quantum sensing with a single electron spin using real-time adaptive measurements

Science.gov (United States)

Bonato, C.; Blok, M. S.; Dinani, H. T.; Berry, D. W.; Markham, M. L.; Twitchen, D. J.; Hanson, R.

2016-03-01

Quantum sensors based on single solid-state spins promise a unique combination of sensitivity and spatial resolution. The key challenge in sensing is to achieve minimum estimation uncertainty within a given time and with high dynamic range. Adaptive strategies have been proposed to achieve optimal performance, but their implementation in solid-state systems has been hindered by the demanding experimental requirements. Here, we realize adaptive d.c. sensing by combining single-shot readout of an electron spin in diamond with fast feedback. By adapting the spin readout basis in real time based on previous outcomes, we demonstrate a sensitivity in Ramsey interferometry surpassing the standard measurement limit. Furthermore, we find by simulations and experiments that adaptive protocols offer a distinctive advantage over the best known non-adaptive protocols when overhead and limited estimation time are taken into account. Using an optimized adaptive protocol we achieve a magnetic field sensitivity of 6.1 ± 1.7 nT Hz-1/2 over a wide range of 1.78 mT. These results open up a new class of experiments for solid-state sensors in which real-time knowledge of the measurement history is exploited to obtain optimal performance.

Real time drift measurement for colloidal probe atomic force microscope: a visual sensing approach

Energy Technology Data Exchange (ETDEWEB)

Wang, Yuliang, E-mail: wangyuliang@buaa.edu.cn; Bi, Shusheng [Robotics Institute, School of Mechanical Engineering and Automation, Beihang University, Beijing 100191 (China); Wang, Huimin [Department of Materials Science and Engineering, The Ohio State University, 2041 College Rd., Columbus, OH 43210 (United States)

2014-05-15

Drift has long been an issue in atomic force microscope (AFM) systems and limits their ability to make long time period measurements. In this study, a new method is proposed to directly measure and compensate for the drift between AFM cantilevers and sample surfaces in AFM systems. This was achieved by simultaneously measuring z positions for beads at the end of an AFM colloidal probe and on sample surface through an off-focus image processing based visual sensing method. The working principle and system configuration are presented. Experiments were conducted to validate the real time drift measurement and compensation. The implication of the proposed method for regular AFM measurements is discussed. We believe that this technique provides a practical and efficient approach for AFM experiments requiring long time period measurement.

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.

Practical Testing and Performance Analysis of Phasor Measurement Unit Using Real Time Digital Simulator (RTDS)

DEFF Research Database (Denmark)

Liu, Leo; Rather, Zakir Hussain; Stearn, Nathen

2012-01-01

Wide Area Measurement Systems (WAMS) and Wide Area Monitoring, Protection and Control Systems (WAMPACS) have evolved rapidly over the last two decades [1]. This fast emerging technology enables real time synchronized monitoring of power systems. Presently, WAMS are mainly used for real time...... visualisation and post event analysis of power systems. It is expected however, that through integration with traditional Supervisory Control and Data Acquisition (SCADA) systems, closed loop control applications will be possible. Phasor Measurement Units (PMUs) are fundamental components of WAMS. Large WAMS...... proposed to realize highly precise phasoreasurements. Further a comparative study based on features of PMUs from different major manufacturers is presented. The selection of optimal parameters, such as phasor format and filter length is also discussed for various applications....

Magnetostrictive patch sensor system for battery-less real-time measurement of torsional vibrations of rotating shafts

Science.gov (United States)

Lee, Jun Kyu; Seung, Hong Min; Park, Chung Il; Lee, Joo Kyung; Lim, Do Hyeong; Kim, Yoon Young

2018-02-01

Real-time uninterrupted measurement for torsional vibrations of rotating shafts is crucial for permanent health monitoring. So far, strain gauge systems with telemetry units have been used for real-time monitoring. However, they have a critical disadvantage in that shaft operations must be stopped intermittently to replace telemetry unit batteries. To find an alternative method to carry out battery-less real-time measurement for torsional vibrations of rotating shafts, a magnetostrictive patch sensor system was proposed in the present study. Since the proposed sensor does not use any powered telemetry system, no battery is needed and thus there is no need to stop rotating shafts for battery replacement. The proposed sensor consists of magnetostrictive patches and small magnets tightly bonded onto a shaft. A solenoid coil is placed around the shaft to convert magnetostrictive patch deformation by shaft torsional vibration into electric voltage output. For sensor design and characterization, investigations were performed in a laboratory on relatively small-sized stationary solid shaft. A magnetostrictive patch sensor system was then designed and installed on a large rotating propulsion shaft of an LPG carrier ship in operation. Vibration signals were measured using the proposed sensor system and compared to those measured with a telemetry unit-equipped strain gauge system.

A Practical Framework to Study Low-Power Scheduling Algorithms on Real-Time and Embedded Systems

Directory of Open Access Journals (Sweden)

Jian (Denny Lin

2014-05-01

Full Text Available With the advanced technology used to design VLSI (Very Large Scale Integration circuits, low-power and energy-efficiency have played important roles for hardware and software implementation. Real-time scheduling is one of the fields that has attracted extensive attention to design low-power, embedded/real-time systems. The dynamic voltage scaling (DVS and CPU shut-down are the two most popular techniques used to design the algorithms. In this paper, we firstly review the fundamental advances in the research of energy-efficient, real-time scheduling. Then, a unified framework with a real Intel PXA255 Xscale processor, namely real-energy, is designed, which can be used to measure the real performance of the algorithms. We conduct a case study to evaluate several classical algorithms by using the framework. The energy efficiency and the quantitative difference in their performance, as well as the practical issues found in the implementation of these algorithms are discussed. Our experiments show a gap between the theoretical and real results. Our framework not only gives researchers a tool to evaluate their system designs, but also helps them to bridge this gap in their future works.

A quantitative real time polymerase chain reaction approach for estimating processed animal proteins in feed: preliminary data

Directory of Open Access Journals (Sweden)

Maria Cesarina Abete

2013-04-01

Full Text Available Lifting of the ban on the use of processed animal proteins (PAPs from non-ruminants in non-ruminant feed is in the wind, avoiding intraspecies recycling. Discrimination of species will be performed through polymerase chain reaction (PCR, which is at a moment a merely qualitative method. Nevertheless, quantification of PAPs in feed is needed. The aim of this study was to approach the quantitative determination of PAPs in feed through Real Time (RT-PCR technique; three different protocols picked up from the literature were tested. Three different kind of matrices were examined: pure animal meals (bovine, chicken and pork; one feed sample certified by the European reference laboratory on animal proteins (EURL AP in feed spiked with 0.1% bovine meal; and genomic DNAs from bovine, chicken and pork muscles. The limit of detection (LOD of the three protocols was set up. All the results obtained from the three protocols considered failed in the quantification process, most likely due to the uncertain copy numbers of the analytical targets chosen. This preliminary study will allow us to address further investigations, with the purpose of developing a RT-PCR quantitative method.

Using Biometric Measurement in Real-Time as a Sympathetic System in Computer Games

Science.gov (United States)

Charij, Stephanie; Oikonomou, Andreas

2013-01-01

With the increasing potential for gaming hardware and peripherals to support biometrics, their application within the games industry for software and design should be considered. This paper assesses the ability to use a form of biometric measurement, heart rate, in real-time to improve the challenge and enjoyment of a game by catering it to…

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.

Study of building materials impregnation processes by quasi-real-time neutron radiography

International Nuclear Information System (INIS)

Nemec, T.; Rant, J.; Apih, V.; Glumac, B.

1999-01-01

Neutron radiography (NR) is a useful non-destructive method for determination of hydrogen content in various building and technical materials. Monitoring of transport processes of moisture and hydrogenous liquids in porous building materials is enabled by fast, quasi-real-time NR methods based on novel imaging plate neutron detectors (IP-NDs). Hydrogen content in the samples is determined by quantitative analysis of measured profiles of neutron attenuation in the samples. Detailed description of quantitative NR method is presented by the authors in another accompanying contribution at this conference. Deterioration of building materials is originated by different processes that all require presence of water therefore it is essential to limit or prevent the transport of water through the porous material. In this presentation, results of a study of clay brick impregnation by silicone based hydrophobic agents will be presented. Quantitative results obtained by NR imaging successfully explained the processes that occur during the impregnation of porous materials. Efficiency of hydrophobic treatment was quantitatively evaluated

Impact of real-time measurements for data assimilation in reservoir simulation

Energy Technology Data Exchange (ETDEWEB)

Schulze-Riegert, R; Krosche, M [Scandpower Petroleum Technology GmbH, Hamburg (Germany); Pajonk, O [TU Braunschweig (Germany). Inst. fuer Wissenschaftliches Rechnen; Myrland, T [Morges Teknisk-Naturvitenskapelige Univ. (NTNU), Trondheim (Germany)

2008-10-23

This paper gives an overview on the conceptual background of data assimilation techniques. The framework of sequential data assimilation as described for the ensemble Kalman filter implementation allows a continuous integration of new measurement data. The initial diversity of ensemble members will be critical for the assimilation process and the ability to successfully assimilate measurement data. At the same time the initial ensemble will impact the propagation of uncertainties with crucial consequences for production forecasts. Data assimilation techniques have complimentary features compared to other optimization techniques built on selection or regression schemes. Specifically, EnKF is applicable to real field cases and defines an important perspective for facilitating continuous reservoir simulation model updates in a reservoir life cycle. (orig.)

Practical performance of real-time shot-noise measurement in continuous-variable quantum key distribution

Science.gov (United States)

Wang, Tao; Huang, Peng; Zhou, Yingming; Liu, Weiqi; Zeng, Guihua

2018-01-01

In a practical continuous-variable quantum key distribution (CVQKD) system, real-time shot-noise measurement (RTSNM) is an essential procedure for preventing the eavesdropper exploiting the practical security loopholes. However, the performance of this procedure itself is not analyzed under the real-world condition. Therefore, we indicate the RTSNM practical performance and investigate its effects on the CVQKD system. In particular, due to the finite-size effect, the shot-noise measurement at the receiver's side may decrease the precision of parameter estimation and consequently result in a tight security bound. To mitigate that, we optimize the block size for RTSNM under the ensemble size limitation to maximize the secure key rate. Moreover, the effect of finite dynamics of amplitude modulator in this scheme is studied and its mitigation method is also proposed. Our work indicates the practical performance of RTSNM and provides the real secret key rate under it.

QUANTIFICATION OF ENTEROVIRUS AND HEPATITIS A VIRUSES IN WELLS AND SPRINGS IN EAST TENNESSEE USING REAL-TIME REVERSE TRANSCIPTION PCR

Science.gov (United States)

This project involves development, validation testing and application of a fast, efficient method of quantitatively measuring occurrence and concentration of common human viral pathogens, enterovirus and hepatitis A virus, in ground water samples using real-time reverse transcrip...

Real time dose rate measurements with fiber optic probes based on the RL and OSL of beryllium oxide

International Nuclear Information System (INIS)

Teichmann, T.; Sponner, J.; Jakobi, Ch.; Henniger, J.

2016-01-01

This work covers the examination of fiber optical probes based on the radioluminescence and real time optically stimulated luminescence of beryllium oxide. Experiments are carried out to determine the fundamental dosimetric and temporal properties of the system and evaluate its suitability for dose rate measurements in brachytherapy and other applications using non-pulsed radiation fields. For this purpose the responses of the radioluminescence and optically stimulated luminescence signal have been investigated in the dose rate range of 20 mGy/h to 3.6 Gy/h and for doses of 1 mGy up to 6 Gy. Furthermore, a new, efficient analysis procedure, the double phase reference summing, is introduced, leading to a real time optically stimulated luminescence signal. This method allows a complete compensation of the stem effect during the measurement. In contrast to previous works, the stimulation of the 1 mm cylindrical beryllium oxide detectors is performed with a symmetric function during irradiation. The investigated dose rates range from 0.3 to 3.6 Gy/h. The real time optically stimulated luminescence signal of beryllium oxide shows a dependency on both the dose rate and the applied dose. To overcome the problem of dose dependency, further experiments using higher stimulation intensities have to follow. - Highlights: • RL and OSL measurements with BeO extended to low dose (rate) range. • A new method to obtain the real time OSL: Dual Phase Reference Summing. • Real time OSL signal shows both dose and dose rate dependency. • Real time OSL enables a complete discrimination of the stem effect.

Measuring Sea-Ice Motion in the Arctic with Real Time Photogrammetry

Science.gov (United States)

Brozena, J. M.; Hagen, R. A.; Peters, M. F.; Liang, R.; Ball, D.

2014-12-01

The U.S. Naval Research Laboratory, in coordination with other groups, has been collecting sea-ice data in the Arctic off the north coast of Alaska with an airborne system employing a radar altimeter, LiDAR and a photogrammetric camera in an effort to obtain wide swaths of measurements coincident with Cryosat-2 footprints. Because the satellite tracks traverse areas of moving pack ice, precise real-time estimates of the ice motion are needed to fly a survey grid that will yield complete data coverage. This requirement led us to develop a method to find the ice motion from the aircraft during the survey. With the advent of real-time orthographic photogrammetric systems, we developed a system that measures the sea ice motion in-flight, and also permits post-process modeling of sea ice velocities to correct the positioning of radar and LiDAR data. For the 2013 and 2014 field seasons, we used this Real Time Ice Motion Estimation (RTIME) system to determine ice motion using Applanix's Inflight Ortho software with an Applanix DSS439 system. Operationally, a series of photos were taken in the survey area. The aircraft then turned around and took more photos along the same line several minutes later. Orthophotos were generated within minutes of collection and evaluated by custom software to find photo footprints and potential overlap. Overlapping photos were passed to the correlation software, which selects a series of "chips" in the first photo and looks for the best matches in the second photo. The correlation results are then passed to a density-based clustering algorithm to determine the offset of the photo pair. To investigate any systematic errors in the photogrammetry, we flew several flight lines over a fixed point on various headings, over an area of non-moving ice in 2013. The orthophotos were run through the correlation software to find any residual offsets, and run through additional software to measure chip positions and offsets relative to the aircraft

Development of a real-time and quantitative thrombus sensor for an extracorporeal centrifugal blood pump by near-infrared light.

Science.gov (United States)

Sakota, Daisuke; Fujiwara, Tatsuki; Ohuchi, Katsuhiro; Kuwana, Katsuyuki; Yamazaki, Hiroyuki; Kosaka, Ryo; Nishida, Masahiro; Mizuno, Tomohiro; Arai, Hirokuni; Maruyama, Osamu

2018-01-01

We developed an optical thrombus sensor for a monopivot extracorporeal centrifugal blood pump. In this study, we investigated its quantitative performance for thrombus detection in acute animal experiments of left ventricular assist using the pump on pathogen-free pigs. Optical fibers were set in the driver unit of the pump. The incident light at the near-infrared wavelength of 810 nm was aimed at the pivot bearing, and the resulting scattered light was guided to the optical fibers. The detected signal was analyzed to obtain the thrombus formation level. As a result, real-time and quantitative monitoring of the thrombus surface area on the pivot bearing was achieved with an accuracy of 3.6 ± 2.3 mm 2 . In addition, the sensing method using the near-infrared light was not influenced by changes in the oxygen saturation and the hematocrit. It is expected that the developed sensor will be useful for optimal anticoagulation management for long-term extracorporeal circulation therapies.

Real-time interferometric monitoring and measuring of photopolymerization based stereolithographic additive manufacturing process: sensor model and algorithm

International Nuclear Information System (INIS)

Zhao, X; Rosen, D W

2017-01-01

As additive manufacturing is poised for growth and innovations, it faces barriers of lack of in-process metrology and control to advance into wider industry applications. The exposure controlled projection lithography (ECPL) is a layerless mask-projection stereolithographic additive manufacturing process, in which parts are fabricated from photopolymers on a stationary transparent substrate. To improve the process accuracy with closed-loop control for ECPL, this paper develops an interferometric curing monitoring and measuring (ICM and M) method which addresses the sensor modeling and algorithms issues. A physical sensor model for ICM and M is derived based on interference optics utilizing the concept of instantaneous frequency. The associated calibration procedure is outlined for ICM and M measurement accuracy. To solve the sensor model, particularly in real time, an online evolutionary parameter estimation algorithm is developed adopting moving horizon exponentially weighted Fourier curve fitting and numerical integration. As a preliminary validation, simulated real-time measurement by offline analysis of a video of interferograms acquired in the ECPL process is presented. The agreement between the cured height estimated by ICM and M and that measured by microscope indicates that the measurement principle is promising as real-time metrology for global measurement and control of the ECPL process. (paper)

Strain measurement of abdominal aortic aneurysm with real-time 3D ultrasound speckle tracking.

Science.gov (United States)

Bihari, P; Shelke, A; Nwe, T H; Mularczyk, M; Nelson, K; Schmandra, T; Knez, P; Schmitz-Rixen, T

2013-04-01

Abdominal aortic aneurysm rupture is caused by mechanical vascular tissue failure. Although mechanical properties within the aneurysm vary, currently available ultrasound methods assess only one cross-sectional segment of the aorta. This study aims to establish real-time 3-dimensional (3D) speckle tracking ultrasound to explore local displacement and strain parameters of the whole abdominal aortic aneurysm. Validation was performed on a silicone aneurysm model, perfused in a pulsatile artificial circulatory system. Wall motion of the silicone model was measured simultaneously with a commercial real-time 3D speckle tracking ultrasound system and either with laser-scan micrometry or with video photogrammetry. After validation, 3D ultrasound data were collected from abdominal aortic aneurysms of five patients and displacement and strain parameters were analysed. Displacement parameters measured in vitro by 3D ultrasound and laser scan micrometer or video analysis were significantly correlated at pulse pressures between 40 and 80 mmHg. Strong local differences in displacement and strain were identified within the aortic aneurysms of patients. Local wall strain of the whole abdominal aortic aneurysm can be analysed in vivo with real-time 3D ultrasound speckle tracking imaging, offering the prospect of individual non-invasive rupture risk analysis of abdominal aortic aneurysms. Copyright © 2013 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.

Repeated cycles of chemical and physical disinfection and their influence on Mycobacterium avium subsp. paratuberculosis viability measured by propidium monoazide F57 quantitative real time PCR.

Science.gov (United States)

Kralik, Petr; Babak, Vladimir; Dziedzinska, Radka

2014-09-01

Mycobacterium avium subsp. paratuberculosis (MAP) has a high degree of resistance to chemical and physical procedures frequently used for the elimination of other bacteria. Recently, a method for the determination of viability by exposure of MAP to propidium monoazide (PMA) and subsequent real time quantitative PCR (qPCR) was established and found to be comparable with culture. The aim of this study was to apply the PMA qPCR method to determine the impact of increasing concentration or time and repeated cycles of the application of selected disinfectants on MAP viability. Different MAP isolates responded to the same type of stress in different ways. The laboratory strain CAPM 6381 had the highest tolerance, while the 8819 low-passage field isolate was the most sensitive. Ultraviolet exposure caused only a partial reduction in MAP viability; all MAP isolates were relatively resistant to chlorine. Only the application of peracetic acid led to the total elimination of MAP. Repeated application of the treatments resulted in more significant decreases in MAP viability compared to single increases in the concentration or time of exposure to the disinfectant. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

Automated methods for real-time analysis of spent-fuel measurement data

International Nuclear Information System (INIS)

Bosler, G.E.; Rinard, P.M.; Klosterbuer, S.F.; Painter, J.

1988-01-01

Software has been developed for ''real-time'' analysis of neutron and gamma data from GRAND-1/fork measurements on spent-fuel assemblies. Three modules compose the software package. The modules are linked through a database system of files. The first module is part of a general data-base processing code. This module prepares input data files with inventory and correction-factor information for the second module. The second module, called OLAF, operates on a computer attached to the GRAND-1 electronics unit. In this second module, neutron and gamma data from spent-fuel assemblies are analyzed to verify consistency in the facility operator declarations for exposure (burnup) and cooling time. From the analysis, potential discrepancies in the measurement data are questioned while equipment is still installed at the facility and is available for additional measurements. During the measurements, data are written to an output file, called a results file, which can be processed by the third module of the software package. In the third module, printed reports summarizing the data and results are prepared, and neutron and gamma data are written to files that are process by the Deming curve-fitting code

Real-Time Imaging System for the OpenPET

Science.gov (United States)

Tashima, Hideaki; Yoshida, Eiji; Kinouchi, Shoko; Nishikido, Fumihiko; Inadama, Naoko; Murayama, Hideo; Suga, Mikio; Haneishi, Hideaki; Yamaya, Taiga

2012-02-01

The OpenPET and its real-time imaging capability have great potential for real-time tumor tracking in medical procedures such as biopsy and radiation therapy. For the real-time imaging system, we intend to use the one-pass list-mode dynamic row-action maximum likelihood algorithm (DRAMA) and implement it using general-purpose computing on graphics processing units (GPGPU) techniques. However, it is difficult to make consistent reconstructions in real-time because the amount of list-mode data acquired in PET scans may be large depending on the level of radioactivity, and the reconstruction speed depends on the amount of the list-mode data. In this study, we developed a system to control the data used in the reconstruction step while retaining quantitative performance. In the proposed system, the data transfer control system limits the event counts to be used in the reconstruction step according to the reconstruction speed, and the reconstructed images are properly intensified by using the ratio of the used counts to the total counts. We implemented the system on a small OpenPET prototype system and evaluated the performance in terms of the real-time tracking ability by displaying reconstructed images in which the intensity was compensated. The intensity of the displayed images correlated properly with the original count rate and a frame rate of 2 frames per second was achieved with average delay time of 2.1 s.

Real-time gene expression analysis in carp (Cyprinus carpio) skin: inflammatory responses to injury mimicking infection with ectoparasites

NARCIS (Netherlands)

Gonzalez, S.F.; Huising, M.O.; Stakauska, R.; Forlenza, M.; Verburg-van Kemenade, B.M.L.; Buchmann, K.; Nielsen, M.E.; Wiegertjes, G.F.

2007-01-01

We studied a predictive model of gene expression induced by mechanical injury of fish skin, to resolve the confounding effects on the immune system induced by injury and skin parasite-specific molecules. We applied real time quantitative PCR (RQ-PCR) to measure the expression of the pro-inflammatory

Development Status of the WetLab-2 Project: New Tools for On-orbit Real-time Quantitative Gene Expression.

Science.gov (United States)

Jung, Jimmy; Parra, Macarena P.; Almeida, Eduardo; Boone, Travis; Chinn, Tori; Ricco, Antonio; Souza, Kenneth; Hyde, Liz; Rukhsana, Yousuf; Richey, C. Scott

2013-01-01

The primary objective of NASA Ames Research Centers WetLab-2 Project is to place on the ISS a research platform to facilitate gene expression analysis via quantitative real-time PCR (qRT-PCR) of biological specimens grown or cultured on orbit. The WetLab-2 equipment will be capable of processing multiple sample types ranging from microbial cultures to animal tissues dissected on-orbit. In addition to the logistical benefits of in-situ sample processing and analysis, conducting qRT-PCR on-orbit eliminates the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms. The system can also validate terrestrial analyses of samples returned from ISS by providing quantitative on-orbit gene expression benchmarking prior to sample return. The ability to get on orbit data will provide investigators with the opportunity to adjust experimental parameters for subsequent trials based on the real-time data analysis without need for sample return and re-flight. Finally, WetLab-2 can be used for analysis of air, surface, water, and clinical samples to monitor environmental contaminants and crew health. The verification flight of the instrument is scheduled to launch on SpaceX-5 in Aug. 2014.Progress to date: The WetLab-2 project completed a thorough study of commercially available qRT-PCR systems and performed a downselect based on both scientific and engineering requirements. The selected instrument, the Cepheid SmartCycler, has advantages including modular design (16 independent PCR modules), low power consumption, and rapid ramp times. The SmartCycler has multiplex capabilities, assaying up to four genes of interest in each of the 16 modules. The WetLab-2 team is currently working with Cepheid to modify the unit for housing within an EXPRESS rack locker on the ISS. This will enable the downlink of data to the ground and provide uplink capabilities for programming, commanding, monitoring, and instrument maintenance. The project is

Real-time vibration measurement by a spatial phase-shifting technique with a tilted holographic interferogram.

Science.gov (United States)

Nakadate, S; Isshiki, M

1997-01-01

Real-time vibration measurement by a tilted holographic interferogram is presented that utilizes the real-time digital fringe processor of a video signal. Three intensity data sampled at every one-third of the fringe spacing of the tilted fringes are used to calculate the modulation term of the fringe that is a function of a vibration amplitude. A three-dimensional lookup table performs the calculation in a TV repetition rate to give a new fringe profile that contours the vibration amplitude. Vibration modes at the resonant frequencies of a flat speaker were displayed on a monitor as changing the exciting frequency of vibration.

Real time measurement of air radioactivity

International Nuclear Information System (INIS)

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

1998-01-01

A Local Meteorological and Radiological Monitoring System was developed in our institute for several purposes: local monitoring, extending our experience in other location such as Cernavoda NPP and research. This system has meteorological sensors for wind speed and direction, air temperature, solar radiation, relative humidity, rainfall, dose ratemeter (Geiger-Muller counter - TIEX), Alpha-Beta Activity-in-Air Monitor (AB96), Iodine Monitor and Eberline Intelligent Ionization Chamber (FHT 6010). All data are collected by a programmable interface Delta-T Logger that is controlled by a software (ODAS - 'On-line Data Acquisition Software'). ODAS was developed in IFIN-HH. It has the capability to acquire, calculate and transmit real meteorological and radiological data through local network. The developed software controls the interface, the flux of input data through the serial port RS232 and after some processing (system, configuration, input data, connection to the network checking, etc) it creates data files. These files are transmitted on-line to our workstation or in any other place connected to Internet. Data can be collected from the logger at any time during logging. There is no need to stop logging. Data output from the logger can be controlled either from the logger's keypad or from other user terminals. ODAS is operated as follows: - First, the last written file and the date-time of acquired readings are checked. For establishing communication with logger a RS232 level signal must be sent to it. The logger wakes if asleep and sends back RDY signal. Powering the logger may take up to 100 ms to establish a correct RS232 level. Noise on the output lines occurs during this period and communication software may need to take into account such spurious signals. A command must be sent to the logger within 2 s to confirm that the last signal received is real and not spurious. Otherwise, the logger interprets the signal as noise and sleeps. The software sends further

Evaluation of changes in periodontal bacteria in healthy dogs over 6 months using quantitative real-time PCR.

Science.gov (United States)

Maruyama, N; Mori, A; Shono, S; Oda, H; Sako, T

2018-03-01

Porphyromonas gulae, Tannerella forsythia and Campylobacter rectus are considered dominant periodontal pathogens in dogs. Recently, quantitative real-time PCR (qRT-PCR) methods have been used for absolute quantitative determination of oral bacterial counts. The purpose of the present study was to establish a standardized qRT-PCR procedure to quantify bacterial counts of the three target periodontal bacteria (P. gulae, T. forsythia and C. rectus). Copy numbers of the three target periodontal bacteria were evaluated in 26 healthy dogs. Then, changes in bacterial counts of the three target periodontal bacteria were evaluated for 24 weeks in 7 healthy dogs after periodontal scaling. Analytical evaluation of each self-designed primer indicated acceptable analytical imprecision. All 26 healthy dogs were found to be positive for P. gulae, T. forsythia and C. rectus. Median total bacterial counts (copies/ng) of each target genes were 385.612 for P. gulae, 25.109 for T. forsythia and 5.771 for C. rectus. Significant differences were observed between the copy numbers of the three target periodontal bacteria. Periodontal scaling reduced median copy numbers of the three target periodontal bacteria in 7 healthy dogs. However, after periodontal scaling, copy numbers of all three periodontal bacteria significantly increased over time (pperiodontal bacteria in dogs. Furthermore, the present study has revealed that qRT-PCR method can be considered as a new objective evaluation system for canine periodontal disease. Copyright© by the Polish Academy of Sciences.

Detection of exogenous gene doping of IGF-I by a real-time quantitative PCR assay.

Science.gov (United States)

Zhang, Jin-Ju; Xu, Jing-Feng; Shen, Yong-Wei; Ma, Shi-Jiao; Zhang, Ting-Ting; Meng, Qing-Lin; Lan, Wen-Jun; Zhang, Chun; Liu, Xiao-Mei

2017-07-01

Gene doping can be easily concealed since its product is similar to endogenous protein, making its effective detection very challenging. In this study, we selected insulin-like growth factor I (IGF-I) exogenous gene for gene doping detection. First, the synthetic IGF-I gene was subcloned to recombinant adeno-associated virus (rAAV) plasmid to produce recombinant rAAV2/IGF-I-GFP vectors. Second, in an animal model, rAAV2/IGF-I-GFP vectors were injected into the thigh muscle tissue of mice, and then muscle and blood specimens were sampled at different time points for total DNA isolation. Finally, real-time quantitative PCR was employed to detect the exogenous gene doping of IGF-I. In view of the characteristics of endogenous IGF-I gene sequences, a TaqMan probe was designed at the junction of exons 2 and 3 of IGF-I gene to distinguish it from the exogenous IGF-I gene. In addition, an internal reference control plasmid and its probe were used in PCR to rule out false-positive results through comparison of their threshold cycle (Ct) values. Thus, an accurate exogenous IGF-I gene detection approach was developed in this study. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Improved quantification accuracy for duplex real-time PCR detection of genetically modified soybean and maize in heat processed foods

Directory of Open Access Journals (Sweden)

CHENG Fang

2013-04-01

Full Text Available Real-time PCR technique has been widely used in quantitative GMO detection in recent years.The accuracy of GMOs quantification based on the real-time PCR methods is still a difficult problem,especially for the quantification of high processed samples.To develop the suitable and accurate real-time PCR system for high processed GM samples,we made ameliorations to several real-time PCR parameters,including re-designed shorter target DNA fragment,similar lengths of amplified endogenous and exogenous gene targets,similar GC contents and melting temperatures of PCR primers and TaqMan probes.Also,one Heat-Treatment Processing Model (HTPM was established using soybean flour samples containing GM soybean GTS 40-3-2 to validate the effectiveness of the improved real-time PCR system.Tested results showed that the quantitative bias of GM content in heat processed samples were lowered using the new PCR system.The improved duplex real-time PCR was further validated using processed foods derived from GM soybean,and more accurate GM content values in these foods was also achieved.These results demonstrated that the improved duplex real-time PCR would be quite suitable in quantitative detection of high processed food products.

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.

Use of quantitative real-time PCR for direct detection of serratia marcescens in marine and other aquatic environments.

Science.gov (United States)

Joyner, Jessica; Wanless, David; Sinigalliano, Christopher D; Lipp, Erin K

2014-03-01

Serratia marcescens is the etiological agent of acroporid serratiosis, a distinct form of white pox disease in the threatened coral Acropora palmata. The pathogen is commonly found in untreated human waste in the Florida Keys, which may contaminate both nearshore and offshore waters. Currently there is no direct method for detection of this bacterium in the aquatic or reef environment, and culture-based techniques may underestimate its abundance in marine waters. A quantitative real-time PCR assay was developed to detect S. marcescens directly from environmental samples, including marine water, coral mucus, sponge tissue, and wastewater. The assay targeted the luxS gene and was able to distinguish S. marcescens from other Serratia species with a reliable quantitative limit of detection of 10 cell equivalents (CE) per reaction. The method could routinely discern the presence of S. marcescens for as few as 3 CE per reaction, but it could not be reliably quantified at this level. The assay detected environmental S. marcescens in complex sewage influent samples at up to 761 CE ml(-1) and in septic system-impacted residential canals in the Florida Keys at up to 4.1 CE ml(-1). This detection assay provided rapid quantitative abilities and good sensitivity and specificity, which should offer an important tool for monitoring this ubiquitous pathogen that can potentially impact both human health and coral health.

Real-time probabilistic covariance tracking with efficient model update.

Science.gov (United States)

Wu, Yi; Cheng, Jian; Wang, Jinqiao; Lu, Hanqing; Wang, Jun; Ling, Haibin; Blasch, Erik; Bai, Li

2012-05-01

The recently proposed covariance region descriptor has been proven robust and versatile for a modest computational cost. The covariance matrix enables efficient fusion of different types of features, where the spatial and statistical properties, as well as their correlation, are characterized. The similarity between two covariance descriptors is measured on Riemannian manifolds. Based on the same metric but with a probabilistic framework, we propose a novel tracking approach on Riemannian manifolds with a novel incremental covariance tensor learning (ICTL). To address the appearance variations, ICTL incrementally learns a low-dimensional covariance tensor representation and efficiently adapts online to appearance changes of the target with only O(1) computational complexity, resulting in a real-time performance. The covariance-based representation and the ICTL are then combined with the particle filter framework to allow better handling of background clutter, as well as the temporary occlusions. We test the proposed probabilistic ICTL tracker on numerous benchmark sequences involving different types of challenges including occlusions and variations in illumination, scale, and pose. The proposed approach demonstrates excellent real-time performance, both qualitatively and quantitatively, in comparison with several previously proposed trackers.

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

NARCIS (Netherlands)

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

2003-01-01

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

FPGA-based real-time phase measuring profilometry algorithm design and implementation

Science.gov (United States)

Zhan, Guomin; Tang, Hongwei; Zhong, Kai; Li, Zhongwei; Shi, Yusheng

2016-11-01

Phase measuring profilometry (PMP) has been widely used in many fields, like Computer Aided Verification (CAV), Flexible Manufacturing System (FMS) et al. High frame-rate (HFR) real-time vision-based feedback control will be a common demands in near future. However, the instruction time delay in the computer caused by numerous repetitive operations greatly limit the efficiency of data processing. FPGA has the advantages of pipeline architecture and parallel execution, and it fit for handling PMP algorithm. In this paper, we design a fully pipelined hardware architecture for PMP. The functions of hardware architecture includes rectification, phase calculation, phase shifting, and stereo matching. The experiment verified the performance of this method, and the factors that may influence the computation accuracy was analyzed.

Overview of real-time kernels at the Superconducting Super Collider Laboratory

International Nuclear Information System (INIS)

Low, K.; Acharya, S.; Allen, M.; Faught, E.; Haenni, D.; Kalbfleisch, C.

1991-01-01

The Superconducting Super Collider Laboratory (SSCL) will have many subsystems that will require real-time microprocessor control. Examples of such Sub-systems requiring real-time controls are power supply ramp generators and quench protection monitors for the superconducting magnets. The authors plan on using a commercial multitasking real-time kernel in these systems. These kernels must perform in a consistent, reliable and efficient manner. Actual performance measurements have been conducted on four different kernels, all running on the same hardware platform. The measurements fall into two categories. Throughput measurements covering the 'non-real-time' aspects of the kernel include process creation/termination times, interprocess communication facilities involving messages, semaphores and shared memory and memory allocation/deallocation. Measurements concentrating on real-time response are context switch times, interrupt latencies and interrupt task response

Overview of real-time kernels at the Superconducting Super Collider Laboratory

International Nuclear Information System (INIS)

Low, K.; Acharya, S.; Allen, M.; Faught, E.; Haenni, D.; Kalbfleisch, C.

1991-05-01

The Superconducting Super Collider Laboratory (SSCL) will have many subsystems that will require real-time microprocessor control. Examples of such sub-systems requiring real-time controls are power supply ramp generators and quench protection monitors for the superconducting magnets. We plan on using a commercial multitasking real-time kernel in these systems. These kernels must perform in a consistent, reliable and efficient manner. Actual performance measurements have been conducted on four different kernels, all running on the same hardware platform. The measurements fall into two categories. Throughput measurements covering the ''non-real-time'' aspects of the kernel include process creation/termination times, interprocess communication facilities involving messages, semaphores and shared memory and memory allocation/deallocation. Measurements concentrating on real-time response are context switch times, interrupt latencies and interrupt task response. 6 refs., 2 tabs

A novel method for real-time skin impedance measurement during radiofrequency skin tightening treatments.

Science.gov (United States)

Harth, Yoram; Lischinsky, Daniel

2011-03-01

The thermal effects of monopolar and bipolar radiofrequency (RF) have been proven to be beneficial in skin tightening. Nevertheless, these effects were frequently partial or unpredictable because of the uncontrolled nature of monopolar or unipolar RF and the superficial nature of energy flow for bipolar or tripolar configurations. One of the hypotheses for lack or predictability of efficacy of the first-generation RF therapy skin tightening systems is lack of adaptation of delivered power to differences in individual skin impedance. A novel multisource phase-controlled system was used (1 MHz, power range 0-65 W) for treatment and real-time skin impedance measurements in 24 patients (EndyMed PRO™; EndyMed, Cesarea, Israel). This system allows continuous real-time measurement of skin impedance delivering constant energy to the patient skin independent of changes in its impedance. More than 6000 unique skin impedance measurements on 22 patients showed an average session impedance range was 215-584 Ohm with an average of 369 Ohm (standard deviation of 49 Ohm). Analyzing individual pulses (total of 600 readings) showed a significant decrease in impedance during the pulse. These findings validate the expected differences in skin impedance between individual patients and in the same patients during the treatment pulse. Clinical study on 30 patients with facial skin aging using the device has shown high predictability of efficacy (86.7% of patients had good results or better at 3 months' follow-up [decrease of 2 or more grades in Fitzpatrick's wrinkle scale]). The real-time customization of energy according to skin impedance allows a significantly more accurate and safe method of nonablative skin tightening with more consistent and predictable results. © 2011 Wiley Periodicals, Inc.

Selecting a set of housekeeping genes for quantitative real-time PCR in normal and tetraploid haemocytes of soft-shell clams, Mya arenaria.

Science.gov (United States)

Siah, A; Dohoo, C; McKenna, P; Delaporte, M; Berthe, F C J

2008-09-01

The transcripts involved in the molecular mechanisms of haemic neoplasia in relation to the haemocyte ploidy status of the soft-shell clam, Mya arenaria, have yet to be identified. For this purpose, real-time quantitative RT-PCR constitutes a sensitive and efficient technique, which can help determine the gene expression involved in haemocyte tetraploid status in clams affected by haemic neoplasia. One of the critical steps in comparing transcription profiles is the stability of selected housekeeping genes, as well as an accurate normalization. In this study, we selected five reference genes, S18, L37, EF1, EF2 and actin, generally used as single control genes. Their expression was analyzed by real-time quantitative RT-PCR at different levels of haemocyte ploidy status in order to select the most stable genes. Using the geNorm software, our results showed that L37, EF1 and S18 represent the most stable gene expressions related to various ploidy status ranging from 0 to 78% of tetraploid haemocytes in clams sampled in North River (Prince Edward Island, Canada). However, actin gene expression appeared to be highly regulated. Hence, using it as a housekeeping gene in tetraploid haemocytes can result in inaccurate data. To compare gene expression levels related to haemocyte ploidy status in Mya arenaria, using L37, EF1 and S18 as housekeeping genes for accurate normalization is therefore recommended.

Quantitative Real-Time PCR Fecal Source Identification in the ...

Science.gov (United States)

Rivers in the Tillamook Basin play a vital role in supporting a thriving dairy and cheese-making industry, as well as providing a safe water resource for local human and wildlife populations. Historical concentrations of fecal bacteria in these waters are at times too high to allow for safe use leading to economic loss, endangerment of local wildlife, and poor conditions for recreational use. In this study, we employ host-associated qPCR methods for human (HF183/BacR287 and HumM2), ruminant (Rum2Bac), cattle (CowM2 and CowM3), canine (DG3 and DG37), and avian (GFD) fecal pollution combined with high-resolution geographic information system (GIS) land use data and general indicator bacteria measurements to elucidatewater quality spatial and temporal trends. Water samples (n=584) were collected over a 1-year period at 29 sites along the Trask, Kilchis, and Tillamook rivers and tributaries (Tillamook Basin, OR). A total of 16.6% of samples (n=97) yielded E. coli levels considered impaired based on Oregon Department of Environmental Quality bacteria criteria (406 MPN/100mL). Hostassociated genetic indicators were detected at frequencies of 39.2% (HF183/BacR287), 16.3% (HumM2), 74.6% (Rum2Bac), 13.0% (CowM2), 26.7% (CowM3), 19.8% (DG3), 3.2% (DG37), and 53.4% (GFD) across all water samples (n=584). Seasonal trends in avian, cattle, and human fecal pollution sources were evident over the study area. On a sample site basis, quantitative fecal source identification and

Real-time Social Internet Data to Guide Forecasting Models

Energy Technology Data Exchange (ETDEWEB)

Del Valle, Sara Y. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

2016-09-20

Our goal is to improve decision support by monitoring and forecasting events using social media, mathematical models, and quantifying model uncertainty. Our approach is real-time, data-driven forecasts with quantified uncertainty: Not just for weather anymore. Information flow from human observations of events through an Internet system and classification algorithms is used to produce quantitatively uncertain forecast. In summary, we want to develop new tools to extract useful information from Internet data streams, develop new approaches to assimilate real-time information into predictive models, validate approaches by forecasting events, and our ultimate goal is to develop an event forecasting system using mathematical approaches and heterogeneous data streams.

Measuring Performance of Soft Real-Time Tasks on Multi-core Systems

OpenAIRE

Rafiq, Salman

2011-01-01

Multi-core platforms are well established, and they are slowly moving into the area of embedded and real-time systems. Nowadays to take advantage of multi-core systems in terms of throughput, soft real-time applications are run together with general purpose applications under an operating system such as Linux. But due to shared hardware resources in multi-core architectures, it is likely that these applications will interfere and compete with each other. This can cause slower response times f...

GETPrime: a gene- or transcript-specific primer database for quantitative real-time PCR.

Science.gov (United States)

Gubelmann, Carine; Gattiker, Alexandre; Massouras, Andreas; Hens, Korneel; David, Fabrice; Decouttere, Frederik; Rougemont, Jacques; Deplancke, Bart

2011-01-01

The vast majority of genes in humans and other organisms undergo alternative splicing, yet the biological function of splice variants is still very poorly understood in large part because of the lack of simple tools that can map the expression profiles and patterns of these variants with high sensitivity. High-throughput quantitative real-time polymerase chain reaction (qPCR) is an ideal technique to accurately quantify nucleic acid sequences including splice variants. However, currently available primer design programs do not distinguish between splice variants and also differ substantially in overall quality, functionality or throughput mode. Here, we present GETPrime, a primer database supported by a novel platform that uniquely combines and automates several features critical for optimal qPCR primer design. These include the consideration of all gene splice variants to enable either gene-specific (covering the majority of splice variants) or transcript-specific (covering one splice variant) expression profiling, primer specificity validation, automated best primer pair selection according to strict criteria and graphical visualization of the latter primer pairs within their genomic context. GETPrime primers have been extensively validated experimentally, demonstrating high transcript specificity in complex samples. Thus, the free-access, user-friendly GETPrime database allows fast primer retrieval and visualization for genes or groups of genes of most common model organisms, and is available at http://updepla1srv1.epfl.ch/getprime/. Database URL: http://deplanckelab.epfl.ch.

Quantification of Human Fecal Bifidobacterium Species by Use of Quantitative Real-Time PCR Analysis Targeting the groEL Gene

Science.gov (United States)

Junick, Jana

2012-01-01

Quantitative real-time PCR assays targeting the groEL gene for the specific enumeration of 12 human fecal Bifidobacterium species were developed. The housekeeping gene groEL (HSP60 in eukaryotes) was used as a discriminative marker for the differentiation of Bifidobacterium adolescentis, B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. dentium, B. gallicum, B. longum, B. pseudocatenulatum, B. pseudolongum, and B. thermophilum. The bifidobacterial chromosome contains a single copy of the groEL gene, allowing the determination of the cell number by quantification of the groEL copy number. Real-time PCR assays were validated by comparing fecal samples spiked with known numbers of a given Bifidobacterium species. Independent of the Bifidobacterium species tested, the proportion of groEL copies recovered from fecal samples spiked with 5 to 9 log10 cells/g feces was approximately 50%. The quantification limit was 5 to 6 log10 groEL copies/g feces. The interassay variability was less than 10%, and variability between different DNA extractions was less than 23%. The method developed was applied to fecal samples from healthy adults and full-term breast-fed infants. Bifidobacterial diversity in both adults and infants was low, with mostly ≤3 Bifidobacterium species and B. longum frequently detected. The predominant species in infant and adult fecal samples were B. breve and B. adolescentis, respectively. It was possible to distinguish B. catenulatum and B. pseudocatenulatum. We conclude that the groEL gene is a suitable molecular marker for the specific and accurate quantification of human fecal Bifidobacterium species by real-time PCR. PMID:22307308

Real time magnetic field and flux measurements for tokamak control using a multi-core PCI Express system

International Nuclear Information System (INIS)

Giannone, L.; Schneider, W.; McCarthy, P.J.; Sips, A.C.C.; Treutterer, W.; Behler, K.; Eich, T.; Fuchs, J.C.; Hicks, N.; Kallenbach, A.; Maraschek, M.; Mlynek, A.; Neu, G.; Pautasso, G.; Raupp, G.; Reich, M.; Schuhbeck, K.H.; Stober, J.; Volpe, F.; Zehetbauer, T.

2009-01-01

The existing real time system for the position and shape control in ASDEX Upgrade has been extended to calculate magnetic flux surfaces in real time using a multi-core PCI Express system running LabVIEW RT. The availability of reflective memory for LabVIEW RT will allow this system to be connected to the control system and other diagnostics in a multi-platform real time network. The measured response of each magnetic probe to the individual poloidal field coil currents in the absence of plasma current is compared to the calculated value. Prior to a tokamak discharge this comparison can be used to check for failure of the magnetic probe, flux loop or integrator.

Multivariate performance reliability prediction in real-time

International Nuclear Information System (INIS)

Lu, S.; Lu, H.; Kolarik, W.J.

2001-01-01

This paper presents a technique for predicting system performance reliability in real-time considering multiple failure modes. The technique includes on-line multivariate monitoring and forecasting of selected performance measures and conditional performance reliability estimates. The performance measures across time are treated as a multivariate time series. A state-space approach is used to model the multivariate time series. Recursive forecasting is performed by adopting Kalman filtering. The predicted mean vectors and covariance matrix of performance measures are used for the assessment of system survival/reliability with respect to the conditional performance reliability. The technique and modeling protocol discussed in this paper provide a means to forecast and evaluate the performance of an individual system in a dynamic environment in real-time. The paper also presents an example to demonstrate the technique

Strain ratio measurement of femoral cartilage by real-time elastosonography: preliminary results

International Nuclear Information System (INIS)

Ipek, Ali; Unal, Ozlem; Kartal, Merve Gulbiz; Arslan, Halil; Isik, Cetin; Bozkurt, Murat

2015-01-01

The purpose of this study was to evaluate strain ratio measurement of femoral cartilage using real-time elastosonography. Twenty-five patients with femoral cartilage pathology on MRI (study group) were prospectively compared with 25 subjects with normal findings on MRI (control group) using real-time elastosonography. Strain ratio measurements of pathologic and normal cartilage were performed and compared, both within the study group and between the two groups. Elastosonography colour-scale coding showed a colour change from blue to red in pathologic cartilage and only blue colour-coding in normal cartilage. In the study group, the median strain ratio was higher in pathologic cartilage areas compared to normal areas (median, 1.49 [interquartile range, 0.80-2.53] vs. median, 0.01 [interquartile range, 0.01-0.01], p < 0.001, respectively). The median strain ratio of the control group was 0.01 (interquartile range, 0.01-0.01), and there was no significant difference compared to normal areas of the study group. There was, however, a significant difference between the control group cartilage and pathologic cartilage of the study group (p < 0.001). Elastosonography may be an effective, easily accessible, and relatively simple tool to demonstrate pathologic cartilage and to differentiate it from normal cartilage in the absence of advanced imaging facility such as MRI. (orig.)

Real-time three-dimensional surface measurement by color encoded light projection

International Nuclear Information System (INIS)

Chen, S. Y.; Li, Y. F.; Guan, Q.; Xiao, G.

2006-01-01

Existing noncontact methods for surface measurement suffer from the disadvantages of poor reliability, low scanning speed, or high cost. The authors present a method for real-time three-dimensional data acquisition by a color-coded vision sensor composed of common components. The authors use a digital projector controlled by computer to generate desired color light patterns. The unique indexing of the light codes is a key problem and is solved in this study so that surface perception can be performed with only local pattern analysis of the neighbor color codes in a single image. Experimental examples and performance analysis are provided

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.

Continuous Real-time Measurements of Vertical Distribution of Magnetic Susceptibility In Soils

Science.gov (United States)

Petrovsky, E.; Hulka, Z.; Kapicka, A.; Magprox Team

Measurements of top-soil magnetic susceptibility are used in approximative outlining polluted areas. However, one of the serious limitations of the method is discrimina- tion between top-soil layers enhanced by atmospherically deposited anthropogenic particles from those dominated by natural particles migrating from magnetically-rich basement rocks. For this purpose, measurements of vertical distribution of magnetic susceptibility along soil profiles is one of the most effective ways in estimating the effect of lithogenic contribution. Up to now, in most cases soil cores have to be mea- sured in laboratory. This method is quite time consuming and does not allow flexible decision about the suitability of the measured site for surface magnetic mapping. In our contribution we will present a new device enabling continuous real-time measure- ments of vertical distribution of magnetic susceptibility directly in field, performed in holes after soil coring. The method is fast, yielding smooth curves (6 data points per 1 mm dept), at least as sensitive as laboratory methods available until now, and at- tached notebook enables direct, on-line control of the lithogenic versus anthropogenic contributions.

Evaluation of quantitative PCR measurement of bacterial colonization of epithelial cells.

Science.gov (United States)

Schmidt, Marcin T; Olejnik-Schmidt, Agnieszka K; Myszka, Kamila; Borkowska, Monika; Grajek, Włodzimierz

2010-01-01

Microbial colonization is an important step in establishing pathogenic or probiotic relations to host cells and in biofilm formation on industrial or medical devices. The aim of this work was to verify the applicability of quantitative PCR (Real-Time PCR) to measure bacterial colonization of epithelial cells. Salmonella enterica and Caco-2 intestinal epithelial cell line was used as a model. To verify sensitivity of the assay a competition of the pathogen cells to probiotic microorganism was tested. The qPCR method was compared to plate count and radiolabel approach, which are well established techniques in this area of research. The three methods returned similar results. The best quantification accuracy had radiolabel method, followed by qPCR. The plate count results showed coefficient of variation two-times higher than this of qPCR. The quantitative PCR proved to be a reliable method for enumeration of microbes in colonization assay. It has several advantages that make it very useful in case of analyzing mixed populations, where several different species or even strains can be monitored at the same time.

Evaluation of methods for oligonucleotide array data via quantitative real-time PCR.

Science.gov (United States)

Qin, Li-Xuan; Beyer, Richard P; Hudson, Francesca N; Linford, Nancy J; Morris, Daryl E; Kerr, Kathleen F

2006-01-17

There are currently many different methods for processing and summarizing probe-level data from Affymetrix oligonucleotide arrays. It is of great interest to validate these methods and identify those that are most effective. There is no single best way to do this validation, and a variety of approaches is needed. Moreover, gene expression data are collected to answer a variety of scientific questions, and the same method may not be best for all questions. Only a handful of validation studies have been done so far, most of which rely on spike-in datasets and focus on the question of detecting differential expression. Here we seek methods that excel at estimating relative expression. We evaluate methods by identifying those that give the strongest linear association between expression measurements by array and the "gold-standard" assay. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) is generally considered the "gold-standard" assay for measuring gene expression by biologists and is often used to confirm findings from microarray data. Here we use qRT-PCR measurements to validate methods for the components of processing oligo array data: background adjustment, normalization, mismatch adjustment, and probeset summary. An advantage of our approach over spike-in studies is that methods are validated on a real dataset that was collected to address a scientific question. We initially identify three of six popular methods that consistently produced the best agreement between oligo array and RT-PCR data for medium- and high-intensity genes. The three methods are generally known as MAS5, gcRMA, and the dChip mismatch mode. For medium- and high-intensity genes, we identified use of data from mismatch probes (as in MAS5 and dChip mismatch) and a sequence-based method of background adjustment (as in gcRMA) as the most important factors in methods' performances. However, we found poor reliability for methods using mismatch probes for low-intensity genes

Evaluation of methods for oligonucleotide array data via quantitative real-time PCR

Directory of Open Access Journals (Sweden)

Morris Daryl E

2006-01-01

Full Text Available Abstract Background There are currently many different methods for processing and summarizing probe-level data from Affymetrix oligonucleotide arrays. It is of great interest to validate these methods and identify those that are most effective. There is no single best way to do this validation, and a variety of approaches is needed. Moreover, gene expression data are collected to answer a variety of scientific questions, and the same method may not be best for all questions. Only a handful of validation studies have been done so far, most of which rely on spike-in datasets and focus on the question of detecting differential expression. Here we seek methods that excel at estimating relative expression. We evaluate methods by identifying those that give the strongest linear association between expression measurements by array and the "gold-standard" assay. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR is generally considered the "gold-standard" assay for measuring gene expression by biologists and is often used to confirm findings from microarray data. Here we use qRT-PCR measurements to validate methods for the components of processing oligo array data: background adjustment, normalization, mismatch adjustment, and probeset summary. An advantage of our approach over spike-in studies is that methods are validated on a real dataset that was collected to address a scientific question. Results We initially identify three of six popular methods that consistently produced the best agreement between oligo array and RT-PCR data for medium- and high-intensity genes. The three methods are generally known as MAS5, gcRMA, and the dChip mismatch mode. For medium- and high-intensity genes, we identified use of data from mismatch probes (as in MAS5 and dChip mismatch and a sequence-based method of background adjustment (as in gcRMA as the most important factors in methods' performances. However, we found poor reliability for methods

Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment

Directory of Open Access Journals (Sweden)

Qinghua Qiu

2018-04-01

Full Text Available Background Freemartinism generally occurs in female offspring of dizygotic twins in a mixed-sex pregnancy. Most bovine heterosexual twin females are freemartins. However, about 10% of bovine heterosexual twin females are fertile. Farmers mostly cull bovine fertile heterosexual twin females due to the lack of a practical diagnostic approach. Culling of such animals results in economic and genetic-material losses both for dairy and beef industry. Methods In this study, a comparative test, including qualitative detection of SRY gene by polymerase chain reaction (PCR, quantitative detection of relative content of SRY by real-time quantitative polymerase chain reaction (qPCR, and quantitative detection of H-Y antigen, was performed to establish the most accurate diagnosis for freemartin. Twelve Holstein heterosexual twin females were used in this study, while three normal Holstein bulls and three normal Holstein cows were used as a positive and negative control, respectively. Results Polymerase chain reaction results revealed that SRY gene were absent in three heterosexual twin females and only two of them were verified as fertile in later age. The qPCR results showed that relative content of SRY was more than 14.2% in freemartins and below 0.41% in fertile heterosexual twin females. The H-Y antigen test showed no significant numerical difference between freemartin and fertile heterosexual twin female. Discussion Our results show that relative content of SRY quantified by qPCR is a better detection method for diagnosis of freemartin in Holstein cattle as compare to qualitative detection of SRY gene by PCR or quantitative detection of H-Y antigen. To the authors’ knowledge, this is the first time we applied qPCR to diagnosing freemartin by quantifying SRY gene and got relative SRY content of each freemartin and fertile heterosexual twin female. We concluded that low-level of SRY would not influence fertility of bovine heterosexual twin female.

Real-time gene expression analysis in carp (Cyprinus carpio L.) skin: Inflammatory responses to injury mimicking infection with ectoparasites

NARCIS (Netherlands)

Gonzalez, S.F.; Huising, M.O.; Stakauskas, R.; Forlenza, M.; Verburg-van Kemenade, B.M.L.; Buchmann, K.; Nielsen, M.E.; Wiegertjes, G.F.

2007-01-01

We studied a predictive model of gene expression induced by mechanical injury of fish skin, to resolve the confounding effects on the immune system induced by injury and skin parasite-specific molecules. We applied real time quantitative PCR (RQ-PCR) to measure the expression of the pro-inflammatory

Comparison of real-time instruments and gravimetric method when measuring particulate matter in a residential building.

Science.gov (United States)

Wang, Zuocheng; Calderón, Leonardo; Patton, Allison P; Sorensen Allacci, MaryAnn; Senick, Jennifer; Wener, Richard; Andrews, Clinton J; Mainelis, Gediminas

2016-11-01

This study used several real-time and filter-based aerosol instruments to measure PM 2.5 levels in a high-rise residential green building in the Northeastern US and compared performance of those instruments. PM 2.5 24-hr average concentrations were determined using a Personal Modular Impactor (PMI) with 2.5 µm cut (SKC Inc., Eighty Four, PA) and a direct reading pDR-1500 (Thermo Scientific, Franklin, MA) as well as its filter. 1-hr average PM 2.5 concentrations were measured in the same apartments with an Aerotrak Optical Particle Counter (OPC) (model 8220, TSI, Inc., Shoreview, MN) and a DustTrak DRX mass monitor (model 8534, TSI, Inc., Shoreview, MN). OPC and DRX measurements were compared with concurrent 1-hr mass concentration from the pDR-1500. The pDR-1500 direct reading showed approximately 40% higher particle mass concentration compared to its own filter (n = 41), and 25% higher PM 2.5 mass concentration compared to the PMI 2.5 filter. The pDR-1500 direct reading and PMI 2.5 in non-smoking homes (self-reported) were not significantly different (n = 10, R 2 = 0.937), while the difference between measurements for smoking homes was 44% (n = 31, R 2 = 0.773). Both OPC and DRX data had substantial and significant systematic and proportional biases compared with pDR-1500 readings. However, these methods were highly correlated: R 2 = 0.936 for OPC versus pDR-1500 reading and R 2 = 0.863 for DRX versus pDR-1500 reading. The data suggest that accuracy of aerosol mass concentrations from direct-reading instruments in indoor environments depends on the instrument, and that correction factors can be used to reduce biases of these real-time monitors in residential green buildings with similar aerosol properties. This study used several real-time and filter-based aerosol instruments to measure PM 2.5 levels in a high-rise residential green building in the northeastern United States and compared performance of those instruments. The data show that while the use of real-time

Real-Time Unsteady Loads Measurements Using Hot-Film Sensors

Science.gov (United States)

Mangalam, Arun S.; Moes, Timothy R.

2004-01-01

Several flight-critical aerodynamic problems such as buffet, flutter, stall, and wing rock are strongly affected or caused by abrupt changes in unsteady aerodynamic loads and moments. Advanced sensing and flow diagnostic techniques have made possible simultaneous identification and tracking, in realtime, of the critical surface, viscosity-related aerodynamic phenomena under both steady and unsteady flight conditions. The wind tunnel study reported here correlates surface hot-film measurements of leading edge stagnation point and separation point, with unsteady aerodynamic loads on a NACA 0015 airfoil. Lift predicted from the correlation model matches lift obtained from pressure sensors for an airfoil undergoing harmonic pitchup and pitchdown motions. An analytical model was developed that demonstrates expected stall trends for pitchup and pitchdown motions. This report demonstrates an ability to obtain unsteady aerodynamic loads in real time, which could lead to advances in air vehicle safety, performance, ride-quality, control, and health management.

Quantitative Real-Time Fluoroscopy Analysis on Measurement of the Hepatic Arterial Flow During Transcatheter Arterial Chemoembolization of Hepatocellular Carcinoma: Comparison with Quantitative Digital Subtraction Angiography Analysis

Energy Technology Data Exchange (ETDEWEB)

Lin, Yi-Yang; Lee, Rheun-Chuan, E-mail: rclee@vghtpe.gov.tw; Guo, Wan-Yuo, E-mail: wyguo@vghtpe.gov.tw; Chu, Wei-Fa [Taipei Veterans General Hospital, Department of Radiology (China); Wu, Frank Chun-Hsien [Siemens Healthcare Ltd. (China); Gehrisch, Sonja [Siemens Healthcare GmbH (Germany)

2016-11-15

PurposeTo quantify the arterial flow change during transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) using digital subtraction angiography, quantitative color-coding analysis (d-QCA), and real-time subtraction fluoroscopy QCA (f-QCA).Materials and MethodsThis prospective study enrolled 20 consecutive patients with HCC who had undergone TACE via a subsegmental approach between February 2014 and April 2015. The TACE endpoint was a sluggish antegrade tumor-feeding arterial flow. d-QCA and f-QCA were used for determining the relative maximal density time (rT{sub max}) of the selected arteries. The rT{sub max} of the selected arteries was analyzed in d-QCA and f-QCA before and after TACE, and its correlation in both analyses was evaluated.ResultsThe pre- and post-TACE rT{sub max} of the embolized segmental artery in d-QCA and f-QCA were 1.59 ± 0.81 and 2.97 ± 1.80 s (P < 0.001) and 1.44 ± 0.52 and 2.28 ± 1.02 s (P < 0.01), respectively. The rT{sub max} of the proximal hepatic artery did not significantly change during TACE in d-QCA and f-QCA. The Spearman correlation coefficients of the pre- and post-TACE rT{sub max} of the embolized segmental artery between d-QCA and f-QCA were 0.46 (P < 0.05) and 0.80 (P < 0.001). Radiation doses in one series of d-QCA and f-QCA were 140.7 ± 51.5 milligray (mGy) and 2.5 ± 0.7 mGy, respectively.Conclusionsf-QCA can quantify arterial flow changes with a higher temporal resolution and lower radiation dose. Flow quantification of the embolized segmental artery using f-QCA and d-QCA is highly correlated.

Quantitative Real-Time Fluoroscopy Analysis on Measurement of the Hepatic Arterial Flow During Transcatheter Arterial Chemoembolization of Hepatocellular Carcinoma: Comparison with Quantitative Digital Subtraction Angiography Analysis

International Nuclear Information System (INIS)

Lin, Yi-Yang; Lee, Rheun-Chuan; Guo, Wan-Yuo; Chu, Wei-Fa; Wu, Frank Chun-Hsien; Gehrisch, Sonja

2016-01-01

PurposeTo quantify the arterial flow change during transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) using digital subtraction angiography, quantitative color-coding analysis (d-QCA), and real-time subtraction fluoroscopy QCA (f-QCA).Materials and MethodsThis prospective study enrolled 20 consecutive patients with HCC who had undergone TACE via a subsegmental approach between February 2014 and April 2015. The TACE endpoint was a sluggish antegrade tumor-feeding arterial flow. d-QCA and f-QCA were used for determining the relative maximal density time (rT_m_a_x) of the selected arteries. The rT_m_a_x of the selected arteries was analyzed in d-QCA and f-QCA before and after TACE, and its correlation in both analyses was evaluated.ResultsThe pre- and post-TACE rT_m_a_x of the embolized segmental artery in d-QCA and f-QCA were 1.59 ± 0.81 and 2.97 ± 1.80 s (P < 0.001) and 1.44 ± 0.52 and 2.28 ± 1.02 s (P < 0.01), respectively. The rT_m_a_x of the proximal hepatic artery did not significantly change during TACE in d-QCA and f-QCA. The Spearman correlation coefficients of the pre- and post-TACE rT_m_a_x of the embolized segmental artery between d-QCA and f-QCA were 0.46 (P < 0.05) and 0.80 (P < 0.001). Radiation doses in one series of d-QCA and f-QCA were 140.7 ± 51.5 milligray (mGy) and 2.5 ± 0.7 mGy, respectively.Conclusionsf-QCA can quantify arterial flow changes with a higher temporal resolution and lower radiation dose. Flow quantification of the embolized segmental artery using f-QCA and d-QCA is highly correlated.

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)

Securing Real-Time Sessions in an IMS-Based Architecture

Science.gov (United States)

Cennamo, Paolo; Fresa, Antonio; Longo, Maurizio; Postiglione, Fabio; Robustelli, Anton Luca; Toro, Francesco

The emerging all-IP mobile network infrastructures based on 3rd Generation IP Multimedia Subsystem philosophy are characterised by radio access technology independence and ubiquitous connectivity for mobile users. Currently, great focus is being devoted to security issues since most of the security threats presently affecting the public Internet domain, and the upcoming ones as well, are going to be suffered by mobile users in the years to come. While a great deal of research activity, together with standardisation efforts and experimentations, is carried out on mechanisms for signalling protection, very few integrated frameworks for real-time multimedia data protection have been proposed in a context of IP Multimedia Subsystem, and even fewer experimental results based on testbeds are available. In this paper, after a general overview of the security issues arising in an advanced IP Multimedia Subsystem scenario, a comprehensive infrastructure for real-time multimedia data protection, based on the adoption of the Secure Real-Time Protocol, is proposed; then, the development of a testbed incorporating such functionalities, including mechanisms for key management and cryptographic context transfer, and allowing the setup of Secure Real-Time Protocol sessions is presented; finally, experimental results are provided together with quantitative assessments and comparisons of system performances for audio sessions with and without the adoption of the Secure Real-Time Protocol framework.

Selecting and validating reference genes for quantitative real-time PCR in Plutella xylostella (L.).

Science.gov (United States)

You, Yanchun; Xie, Miao; Vasseur, Liette; You, Minsheng

2018-05-01

Gene expression analysis provides important clues regarding gene functions, and quantitative real-time PCR (qRT-PCR) is a widely used method in gene expression studies. Reference genes are essential for normalizing and accurately assessing gene expression. In the present study, 16 candidate reference genes (ACTB, CyPA, EF1-α, GAPDH, HSP90, NDPk, RPL13a, RPL18, RPL19, RPL32, RPL4, RPL8, RPS13, RPS4, α-TUB, and β-TUB) from Plutella xylostella were selected to evaluate gene expression stability across different experimental conditions using five statistical algorithms (geNorm, NormFinder, Delta Ct, BestKeeper, and RefFinder). The results suggest that different reference genes or combinations of reference genes are suitable for normalization in gene expression studies of P. xylostella according to the different developmental stages, strains, tissues, and insecticide treatments. Based on the given experimental sets, the most stable reference genes were RPS4 across different developmental stages, RPL8 across different strains and tissues, and EF1-α across different insecticide treatments. A comprehensive and systematic assessment of potential reference genes for gene expression normalization is essential for post-genomic functional research in P. xylostella, a notorious pest with worldwide distribution and a high capacity to adapt and develop resistance to insecticides.

Redundant and fault-tolerant algorithms for real-time measurement and control systems for weapon equipment.

Science.gov (United States)

Li, Dan; Hu, Xiaoguang

2017-03-01

Because of the high availability requirements from weapon equipment, an in-depth study has been conducted on the real-time fault-tolerance of the widely applied Compact PCI (CPCI) bus measurement and control system. A redundancy design method that uses heartbeat detection to connect the primary and alternate devices has been developed. To address the low successful execution rate and relatively large waste of time slices in the primary version of the task software, an improved algorithm for real-time fault-tolerant scheduling is proposed based on the Basic Checking available time Elimination idle time (BCE) algorithm, applying a single-neuron self-adaptive proportion sum differential (PSD) controller. The experimental validation results indicate that this system has excellent redundancy and fault-tolerance, and the newly developed method can effectively improve the system availability. Copyright © 2017 ISA. Published by Elsevier Ltd. All rights reserved.

Plasma density control in real-time on the COMPASS tokamak

Energy Technology Data Exchange (ETDEWEB)

Janky, F., E-mail: filip.janky.work@gmail.com [Institute of Plasma Physics AS CR, v.v.i., Association EURATOM/IPP.CR, Za Slovankou 3, 182 00 Praha 8 (Czech Republic); Department of Surface and Plasma Science, Faculty of Mathematics and Physics, Charles University in Prague, V Holešovičkách 2, 180 00 Praha 8 (Czech Republic); Hron, M. [Institute of Plasma Physics AS CR, v.v.i., Association EURATOM/IPP.CR, Za Slovankou 3, 182 00 Praha 8 (Czech Republic); Havlicek, J. [Institute of Plasma Physics AS CR, v.v.i., Association EURATOM/IPP.CR, Za Slovankou 3, 182 00 Praha 8 (Czech Republic); Department of Surface and Plasma Science, Faculty of Mathematics and Physics, Charles University in Prague, V Holešovičkách 2, 180 00 Praha 8 (Czech Republic); Varavin, M.; Zacek, F.; Seidl, J.; Panek, R. [Institute of Plasma Physics AS CR, v.v.i., Association EURATOM/IPP.CR, Za Slovankou 3, 182 00 Praha 8 (Czech Republic)

2015-10-15

Highlights: • We fitted length of the chord of the interferometry crossing plasma in the different plasma scenarios. • We add correction to the actual length of the chord of the interferometry according to plasma shape and position in real-time code. • We used this correction to control plasma density in real-time. - Abstract: The electron density on COMPASS is measured using 2 mm microwave interferometer. Interferometer signal is used as an input for the feedback control loop, running under the MARTe real-time framework. Two different threads are used to calculate (fast 50 μs thread) and to control (slow 500 μs thread) the electron density. The interferometer measures a line averaged density along a measurement chord. This paper describes an approach to control the line-averaged electron density in a real-time loop, using a correction to the real plasma shape, the plasma position, and non-linear effects of the electron density measurement at high densities. Newly developed real-time electron density control give COMPASS the chance to control the electron density more accurately which is essential for parametric scans for diagnosticians, for physics experiments and also for achieving plasma scenarios with H-mode.

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...

Detection of tumor markers in prostate cancer and comparison of sensitivity between real time and nested PCR.

Science.gov (United States)

Matsuoka, Takayuki; Shigemura, Katsumi; Yamamichi, Fukashi; Fujisawa, Masato; Kawabata, Masato; Shirakawa, Toshiro

2012-06-27

The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivity by real time PCR and nested PCR. In real time PCR, there was a significant correlation between cell number and the RNA concentration obtained (R(2)=0.9944) for PSA, PSMA, and AR. We found it possible to detect these markers from a single LNCaP cell in both real time and nested PCR. By comparison, nested PCR reached a linear curve in fewer PCR cycles than real time PCR, suggesting that nested PCR may offer PCR results more quickly than real time PCR. In conclusion, nested PCR may offer tumor maker detection in PCa cells more quickly (with fewer PCR cycles) with the same high sensitivity as real time PCR. Further study is necessary to establish and evaluate the best tool for PCa tumor marker detection.

Fully integrated microfluidic measurement system for real-time determination of gas and liquid mixtures composition

NARCIS (Netherlands)

Lötters, Joost Conrad; Groenesteijn, Jarno; van der Wouden, E.J.; Sparreboom, Wouter; Lammerink, Theodorus S.J.; Wiegerink, Remco J.

2015-01-01

We have designed and realised a fully integrated microfluidic measurement system for real-time determination of both flow rate and composition of gas- and liquid mixtures. The system comprises relative permittivity sensors, pressure sensors, a Coriolis flow and density sensor, a thermal flow sensor

[Comparison of two quantitative methods of endobronchial ultrasound real-time elastography for evaluating intrathoracic lymph nodes].

Science.gov (United States)

Mao, X W; Yang, J Y; Zheng, X X; Wang, L; Zhu, L; Li, Y; Xiong, H K; Sun, J Y

2017-06-12

Objective: To compare the clinical value of two quantitative methods in analyzing endobronchial ultrasound real-time elastography (EBUS-RTE) images for evaluating intrathoracic lymph nodes. Methods: From January 2014 to April 2014, EBUS-RTE examination was performed in patients who received EBUS-TBNA examination in Shanghai Chest Hospital. Each intrathoracic lymph node had a selected EBUS-RTE image. Stiff area ratio and mean hue value of region of interest (ROI) in each image were calculated respectively. The final diagnosis of lymph node was based on the pathologic/microbiologic results of EBUS-TBNA, pathologic/microbiologic results of other examinations and clinical following-up. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy were evaluated for distinguishing malignant and benign lesions. Results: Fifty-six patients and 68 lymph nodes were enrolled in this study, of which 35 lymph nodes were malignant and 33 lymph nodes were benign. The stiff area ratio and mean hue value of benign and malignant lesions were 0.32±0.29, 0.62±0.20 and 109.99±28.13, 141.62±17.52, respectively, and statistical differences were found in both of those two methods ( t =-5.14, P methods can be used for analyzing EBUS-RTE images quantitatively, having the value of differentiating benign and malignant intrathoracic lymph nodes, and the stiff area ratio is better than the mean hue value between the two methods.

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...

Near-Real-Time Monitoring of Insect Defoliation Using Landsat Time Series

Directory of Open Access Journals (Sweden)

Valerie J. Pasquarella

2017-07-01

Full Text Available Introduced insects and pathogens impact millions of acres of forested land in the United States each year, and large-scale monitoring efforts are essential for tracking the spread of outbreaks and quantifying the extent of damage. However, monitoring the impacts of defoliating insects presents a significant challenge due to the ephemeral nature of defoliation events. Using the 2016 gypsy moth (Lymantria dispar outbreak in Southern New England as a case study, we present a new approach for near-real-time defoliation monitoring using synthetic images produced from Landsat time series. By comparing predicted and observed images, we assessed changes in vegetation condition multiple times over the course of an outbreak. Initial measures can be made as imagery becomes available, and season-integrated products provide a wall-to-wall assessment of potential defoliation at 30 m resolution. Qualitative and quantitative comparisons suggest our Landsat Time Series (LTS products improve identification of defoliation events relative to existing products and provide a repeatable metric of change in condition. Our synthetic-image approach is an important step toward using the full temporal potential of the Landsat archive for operational monitoring of forest health over large extents, and provides an important new tool for understanding spatial and temporal dynamics of insect defoliators.

In-Line Capacitance Sensor for Real-Time Water Absorption Measurements

Science.gov (United States)

Nurge, Mark A.; Perusich, Stephen A.

2010-01-01

A capacitance/dielectric sensor was designed, constructed, and used to measure in real time the in-situ water concentration in a desiccant water bed. Measurements were carried out with two experimental setups: (1) passing nitrogen through a humidity generator and allowing the gas stream to become saturated at a measured temperature and pressure, and (2) injecting water via a syringe pump into a nitrogen stream. Both water vapor generating devices were attached to a downstream vertically-mounted water capture bed filled with 19.5 g of Moisture Gone desiccant. The sensor consisted of two electrodes: (1) a 1/8" dia stainless steel rod placed in the middle of the bed and (2) the outer shell of the stainless steel bed concentric with the rod. All phases of the water capture process (background, heating, absorption, desorption, and cooling) were monitored with capacitance. The measured capacitance was found to vary linearly with the water content in the bed at frequencies above 100 kHz indicating dipolar motion dominated the signal; below this frequency, ionic motion caused nonlinearities in the water concentration/capacitance relationship. The desiccant exhibited a dielectric relaxation whose activation energy was lowered upon addition of water indicating either a less hindered rotational motion or crystal reorientation.

Quantification of organellar DNA and RNA using real-time PCR.

Science.gov (United States)

Weihe, Andreas

2014-01-01

Quantitative (real-time) polymerase chain reaction (PCR) allows the measurement of relative organellar gene copy numbers as well as transcript abundance of individual mitochondrial or plastidial genes. Requiring only minute amounts of total DNA or RNA, the described method can replace traditional analyses like Southern or Northern hybridization which require large amounts of organellar nucleic acids and usually provide only semiquantitative data. Here we describe prerequisites, reaction conditions, and data analysis principles, which should be applicable for a wide range of plant species and experimental situations where comparative and precise determination of gene copy numbers or transcript abundance is requested. Sequences of amplification primers for qPCR of organellar genes from Arabidopsis are provided.

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

Science.gov (United States)

Zhang, Xin; Zhang, He; Pu, Jinji; Qi, Yanxiang; Yu, Qunfang; Xie, Yixian; Peng, Jun

2013-01-01

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

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

Directory of Open Access Journals (Sweden)

Xin Zhang

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

Real-time high-resolution measurement of collagen alignment in dynamically loaded soft tissue.

Science.gov (United States)

York, Timothy; Kahan, Lindsey; Lake, Spencer P; Gruev, Viktor

2014-06-01

A technique for creating maps of the direction and strength of fiber alignment in collagenous soft tissues is presented. The method uses a division of focal plane polarimeter to measure circularly polarized light transmitted through the tissue. The architecture of the sensor allows measurement of the retardance and fiber alignment at the full frame rate of the sensor without any moving optics. The technique compares favorably to the standard method of using a rotating polarizer. How the new technique enables real-time capture of the full angular spread of fiber alignment and retardance under various cyclic loading conditions is illustrated.

Time-resolved measurements with intense ultrashort laser pulses: a 'molecular movie' in real time

International Nuclear Information System (INIS)

Rudenko, A; Ergler, Th; Feuerstein, B; Zrost, K; Schroeter, C D; Moshammer, R; Ullrich, J

2007-01-01

We report on the high-resolution multidimensional real-time mapping of H 2 + and D 2 + nuclear wave packets performed employing time-resolved three-dimensional Coulomb explosion imaging with intense laser pulses. Exploiting a combination of a 'reaction microscope' spectrometer and a pump-probe setup with two intense 6-7 fs laser pulses, we simultaneously visualize both vibrational and rotational motion of the molecule, and obtain a sequence of snapshots of the squared ro-vibrational wave function with time-step resolution of ∼ 0.3 fs, allowing us to reconstruct a real-time movie of the ultrafast molecular motion. We observe fast dephasing, or 'collapse' of the vibrational wave packet and its subsequent revival, as well as signatures of rotational excitation. For D 2 + we resolve also the fractional revivals resulting from the interference between the counter-propagating parts of the wave packet

Exploring Valid Reference Genes for Quantitative Real-time PCR Analysis in Plutella xylostella (Lepidoptera: Plutellidae)

Science.gov (United States)

Fu, Wei; Xie, Wen; Zhang, Zhuo; Wang, Shaoli; Wu, Qingjun; Liu, Yong; Zhou, Xiaomao; Zhou, Xuguo; Zhang, Youjun

2013-01-01

Abstract: Quantitative real-time PCR (qRT-PCR), a primary tool in gene expression analysis, requires an appropriate normalization strategy to control for variation among samples. The best option is to compare the mRNA level of a target gene with that of reference gene(s) whose expression level is stable across various experimental conditions. In this study, expression profiles of eight candidate reference genes from the diamondback moth, Plutella xylostella, were evaluated under diverse experimental conditions. RefFinder, a web-based analysis tool, integrates four major computational programs including geNorm, Normfinder, BestKeeper, and the comparative ΔCt method to comprehensively rank the tested candidate genes. Elongation factor 1 (EF1) was the most suited reference gene for the biotic factors (development stage, tissue, and strain). In contrast, although appropriate reference gene(s) do exist for several abiotic factors (temperature, photoperiod, insecticide, and mechanical injury), we were not able to identify a single universal reference gene. Nevertheless, a suite of candidate reference genes were specifically recommended for selected experimental conditions. Our finding is the first step toward establishing a standardized qRT-PCR analysis of this agriculturally important insect pest. PMID:23983612

Real-time laser holographic interferometry for aerodynamics

International Nuclear Information System (INIS)

Lee, G.

1987-01-01

Recent developments in thermoplastic recording holograms and advancements in automated image digitalization and analysis make real-time laser holographic interferometry feasible for two-dimensional flows such as airfoil flows. Typical airfoil measurements would include airfoil pressure distributions, wake and boundary layer profiles, and flow field density contours. This paper addresses some of the problems and requirements of a real-time laser holographic interferometer. 13 references

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.

Using quantitative real-time PCR to detect chimeras in transgenic tobacco and apricot and to monitor their dissociation

Directory of Open Access Journals (Sweden)

Burgos Lorenzo

2010-07-01

Full Text Available Abstract Background The routine generation of transgenic plants involves analysis of transgene integration into the host genome by means of Southern blotting. However, this technique cannot distinguish between uniformly transformed tissues and the presence of a mixture of transgenic and non-transgenic cells in the same tissue. On the other hand, the use of reporter genes often fails to accurately detect chimerical tissues because their expression can be affected by several factors, including gene silencing and plant development. So, new approaches based on the quantification of the amount of the transgene are needed urgently. Results We show here that chimeras are a very frequent phenomenon observed after regenerating transgenic plants. Spatial and temporal analyses of transformed tobacco and apricot plants with a quantitative, real-time PCR amplification of the neomycin phosphotransferase (nptII transgene as well as of an internal control (β-actin, used to normalise the amount of target DNA at each reaction, allowed detection of chimeras at unexpected rates. The amount of the nptII transgene differed greatly along with the sub-cultivation period of these plants and was dependent on the localisation of the analysed leaves; being higher in roots and basal leaves, while in the apical leaves it remained at lower levels. These data demonstrate that, unlike the use of the gus marker gene, real-time PCR is a powerful tool for detection of chimeras. Although some authors have proposed a consistent, positive Southern analysis as an alternative methodology for monitoring the dissociation of chimeras, our data show that it does not provide enough proof of uniform transformation. In this work, however, real-time PCR was applied successfully to monitor the dissociation of chimeras in tobacco plants and apricot callus. Conclusions We have developed a rapid and reliable method to detect and estimate the level of chimeras in transgenic tobacco and apricot

A two-domain real-time algorithm for optimal data reduction: A case study on accelerator magnet measurements

CERN Document Server

Arpaia, P; Inglese, V

2010-01-01

A real-time algorithm of data reduction, based on the combination a two lossy techniques specifically optimized for high-rate magnetic measurements in two domains (e.g. time and space), is proposed. The first technique exploits an adaptive sampling rule based on the power estimation of the flux increments in order to optimize the information to be gathered for magnetic field analysis in real time. The tracking condition is defined by the target noise level in the Nyquist band required by post-processing procedure of magnetic analysis. The second technique uses a data reduction algorithm in order to improve the compression ratio while preserving the consistency of the measured signal. The allowed loss is set equal to the random noise level in the signal in order to force the loss and the noise to cancel rather than to add, by improving the signal-to-noise ratio. Numerical analysis and experimental results of on-field performance characterization and validation for two case studies of magnetic measurement syste...

Detection of Tumor Markers in Prostate Cancer and Comparison of Sensitivity between Real Time and Nested PCR

OpenAIRE

Matsuoka, Takayuki; Shigemura, Katsumi; Yamamichi, Fukashi; Fujisawa, Masato; Kawabata, Masato; Shirakawa, Toshiro

2012-01-01

The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivi...

Response of a diuron-degrading community to diuron exposure assessed by real-time quantitative PCR monitoring of phenylurea hydrolase A and B encoding genes

OpenAIRE

Pesce , S.; Beguet , J.; Rouard , N.; Devers Lamrani , M.; Martin Laurent , F.

2013-01-01

A real-time quantitative PCR method was developed to detect and quantify phenlylurea hydrolase genes' (puhA and puhB) sequences from environmental DNA samples to assess diuron-degrading genetic potential in some soil and sediment microbial communities. In the soil communities, mineralization rates (determined with [ring-14C]-labeled diuron) were linked to diuron-degrading genetic potentials estimated from puhB number copies, which increased following repeated diuron treatments. In the sedimen...

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

Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor

DEFF Research Database (Denmark)

Jensen, Pia Wrensted; Falconi, Mattia; Kristoffersen, Emil Laust

2013-01-01

structure of the biosensor. The specific action of TDP1 removes the quencher, thereby enabling optical detection of the fluorophore. Since the enzymatic action of TDP1 is the only “signal amplification” the increase in fluorescence may easily be followed in real-time and allows quantitative analyses of TDP1......Real-time detection of enzyme activities may present the easiest and most reliable way of obtaining quantitative analyses in biological samples. We present a new DNA-biosensor capable of detecting the activity of the potential anticancer drug target tyrosyl-DNA phosphodiesterase 1 (TDP1) in a very...... simple, high throughput, and real-time format. The biosensor is specific for Tdp1 even in complex biological samples, such as human cell extracts, and may consequently find future use in fundamental studies as well as a cancer predictive tool allowing fast analyses of diagnostic cell samples...

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

Applying MDA to SDR for Space to Model Real-time Issues

Science.gov (United States)

Blaser, Tammy M.

2007-01-01

NASA space communications systems have the challenge of designing SDRs with highly-constrained Size, Weight and Power (SWaP) resources. A study is being conducted to assess the effectiveness of applying the MDA Platform-Independent Model (PIM) and one or more Platform-Specific Models (PSM) specifically to address NASA space domain real-time issues. This paper will summarize our experiences with applying MDA to SDR for Space to model real-time issues. Real-time issues to be examined, measured, and analyzed are: meeting waveform timing requirements and efficiently applying Real-time Operating System (RTOS) scheduling algorithms, applying safety control measures, and SWaP verification. Real-time waveform algorithms benchmarked with the worst case environment conditions under the heaviest workload will drive the SDR for Space real-time PSM design.

Exploring valid reference genes for quantitative real-time PCR analysis in Sesamia inferens (Lepidoptera: Noctuidae.

Directory of Open Access Journals (Sweden)

Meng Sun

Full Text Available The pink stem borer, Sesamia inferens, which is endemic in China and other parts of Asia, is a major pest of rice and causes significant yield loss in this host plant. Very few studies have addressed gene expression in S. inferens. Quantitative real-time PCR (qRT-PCR is currently the most accurate and sensitive method for gene expression analysis. In qRT-PCR, data are normalized using reference genes, which help control for internal differences and reduce error between samples. In this study, seven candidate reference genes, 18S ribosomal RNA (18S rRNA, elongation factor 1 (EF1, glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ribosomal protein S13 (RPS13, ribosomal protein S20 (RPS20, tubulin (TUB, and β-actin (ACTB were evaluated for their suitability in normalizing gene expression under different experimental conditions. The results indicated that three genes (RPS13, RPS20, and EF1 were optimal for normalizing gene expression in different insect tissues (head, epidermis, fat body, foregut, midgut, hindgut, Malpighian tubules, haemocytes, and salivary glands. 18S rRNA, EF1, and GAPDH were best for normalizing expression with respect to developmental stages and sex (egg masses; first, second, third, fourth, fifth, and sixth instar larvae; male and female pupae; and one-day-old male and female adults. 18S rRNA, RPS20, and TUB were optimal for fifth instars exposed to different temperatures (-8, -6, -4, -2, 0, and 27°C. To validate this recommendation, the expression profile of a target gene heat shock protein 83 gene (hsp83 was investigated, and results showed the selection was necessary and effective. In conclusion, this study describes reference gene sets that can be used to accurately measure gene expression in S. inferens.

Exploring valid reference genes for quantitative real-time PCR analysis in Sesamia inferens (Lepidoptera: Noctuidae).

Science.gov (United States)

Sun, Meng; Lu, Ming-Xing; Tang, Xiao-Tian; Du, Yu-Zhou

2015-01-01

The pink stem borer, Sesamia inferens, which is endemic in China and other parts of Asia, is a major pest of rice and causes significant yield loss in this host plant. Very few studies have addressed gene expression in S. inferens. Quantitative real-time PCR (qRT-PCR) is currently the most accurate and sensitive method for gene expression analysis. In qRT-PCR, data are normalized using reference genes, which help control for internal differences and reduce error between samples. In this study, seven candidate reference genes, 18S ribosomal RNA (18S rRNA), elongation factor 1 (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein S13 (RPS13), ribosomal protein S20 (RPS20), tubulin (TUB), and β-actin (ACTB) were evaluated for their suitability in normalizing gene expression under different experimental conditions. The results indicated that three genes (RPS13, RPS20, and EF1) were optimal for normalizing gene expression in different insect tissues (head, epidermis, fat body, foregut, midgut, hindgut, Malpighian tubules, haemocytes, and salivary glands). 18S rRNA, EF1, and GAPDH were best for normalizing expression with respect to developmental stages and sex (egg masses; first, second, third, fourth, fifth, and sixth instar larvae; male and female pupae; and one-day-old male and female adults). 18S rRNA, RPS20, and TUB were optimal for fifth instars exposed to different temperatures (-8, -6, -4, -2, 0, and 27°C). To validate this recommendation, the expression profile of a target gene heat shock protein 83 gene (hsp83) was investigated, and results showed the selection was necessary and effective. In conclusion, this study describes reference gene sets that can be used to accurately measure gene expression in S. inferens.

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

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.

Development of Quantitative Competitive PCR and Absolute Based Real-Time PCR Assays for Quantification of The Butyrate Producing Bacterium: Butyrivibrio fibrisolvens

Directory of Open Access Journals (Sweden)

Mojtaba Tahmoorespur

2016-04-01

Full Text Available Introduction Butyrivibrio fibrisolvens strains are presently recognized as the major butyrate-producing bacteria found in the rumen and digestive track of many animals and also in the human gut. In this study we reported the development of two DNA based techniques, quantitative competitive (QC PCR and absolute based Real-Time PCR, for enumerating Butyrivibrio fibrisolvens strains. Despite the recent introduction of real-time PCR method for the rapid quantification of the target DNA sequences, use of quantitative competitive PCR (QC-PCR technique continues to play an important role in nucleic acid quantification since it is more cost effective. The procedure relies on the co-amplification of the sequence of interest with a serially diluted synthetic DNA fragment of the known concentration (competitor, using the single set primers. A real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR. It monitors the amplification of a targeted DNA molecule during the PCR. Materials and Methods At first reported species-specific primers targeting the 16S rDNA region of the bacterium Butyrivibrio fibrisolvens were used for amplifying a 213 bp fragment. A DNA competitor differing by 50 bp in length from the 213 bp fragment was constructed and cloned into pTZ57R/T vector. The competitor was quantified by NanoDrop spectrophotometer and serially diluted and co-amplified by PCR with total extracted DNA from rumen fluid samples. PCR products were quantified by photographing agarose gels and analyzed with Image J software and the amount of amplified target DNA was log plotted against the amount of amplified competitor. Coefficient of determination (R2 was used as a criterion of methodology precision. For developing the Real-time PCR technique, the 213 bp fragment was amplified and cloned into pTZ57R/T was used to draw a standard curve. Results and Discussion The specific primers of Butyrivibrio

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)

Effectiveness of Quantitative Real Time PCR in Long-Term Follow-up of Chronic Myeloid Leukemia Patients.

Science.gov (United States)

Savasoglu, Kaan; Payzin, Kadriye Bahriye; Ozdemirkiran, Fusun; Berber, Belgin

2015-08-01

To determine the use of the Quantitative Real Time PCR (RQ-PCR) assay follow-up with Chronic Myeloid Leukemia (CML) patients. Cross-sectional observational. Izmir Ataturk Education and Research Hospital, Izmir, Turkey, from 2009 to 2013. Cytogenetic, FISH, RQ-PCR test results from 177 CMLpatients' materials selected between 2009 - 2013 years was set up for comparison analysis. Statistical analysis was performed to compare between FISH, karyotype and RQ-PCR results of the patients. Karyotyping and FISH specificity and sensitivity rates determined by ROC analysis compared with RQ-PCR results. Chi-square test was used to compare test failure rates. Sensitivity and specificity values were determined for karyotyping 17.6 - 98% (p=0.118, p > 0.05) and for FISH 22.5 - 96% (p=0.064, p > 0.05) respectively. FISH sensitivity was slightly higher than karyotyping but there was calculated a strong correlation between them (p 0.05); however, karyotyping and FISH test failure rate was statistically significant (p < 0.001). Besides, the situation needed for karyotype analysis, RQ-PCR assay can be used alone in the follow-up of CMLdisease.

High-level expression of podoplanin in benign and malignant soft tissue tumors: immunohistochemical and quantitative real-time RT-PCR analysis.

Science.gov (United States)

Xu, Yongjun; Ogose, Akira; Kawashima, Hiroyuki; Hotta, Tetsuo; Ariizumi, Takashi; Li, Guidong; Umezu, Hajime; Endo, Naoto

2011-03-01

Podoplanin is a 38 kDa mucin-type transmembrane glycoprotein that was first identified in rat glomerular epithelial cells (podocytes). It is expressed in normal lymphatic endothelium, but is absent from vascular endothelial cells. D2-40 is a commercially available mouse monoclonal antibody which binds to an epitope on human podoplanin. D2-40 immunoreactivity is therefore highly sensitive and specific for lymphatic endothelium. Recent investigations have shown widespread applications of immunohistochemical staining with D2-40 in evaluating podoplanin expression as an immunohistochemical marker for diagnosis and prognosis in various tumors. To determine whether the podoplanin (D2-40) antibody may be useful for the diagnosis of soft tissue tumors, 125 cases, including 4 kinds of benign tumors, 15 kinds of malignant tumors and 3 kinds of tumor-like lesions were immunostained using the D2-40 antibody. Total RNA was extracted from frozen tumor tissue obtained from 41 corresponding soft tissue tumor patients and 12 kinds of soft tissue tumor cell lines. Quantitative real-time PCR reactions were performed. Immunohistochemical and quantitative real-time RT-PCR analyses demonstrated the expression of the podoplanin protein and mRNA in the majority of benign and malignant soft tissue tumors and tumor-like lesions examined, with the exception of alveolar soft part sarcoma, embryonal and alveolar rhabdomyosarcoma, extraskeletal Ewing's sarcoma/peripheral primitive neuro-ectodermal tumor and lipoma, which were completely negative for podoplanin. Since it is widely and highly expressed in nearly all kinds of soft tissue tumors, especially in spindle cell sarcoma, myxoid type soft tissue tumors and soft tissue tumors of the nervous system, podoplanin is considered to have little value in the differential diagnosis of soft tissue tumors.

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....

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.)

Quantitation of O6-methylguanine-DNA methyltransferase gene messenger RNA in gliomas by means of real-time RT-PCR and clinical response to nitrosoureas.

Science.gov (United States)

Tanaka, Satoshi; Oka, Hidehiro; Fujii, Kiyotaka; Watanabe, Kaoru; Nagao, Kumi; Kakimoto, Atsushi

2005-09-01

1. O6-methylguanine-DNA methyltransferase (MGMT) mRNA was measured in 50 malignant gliomas that had received 1-(4-amino-2-methyl-5-pyrimidynyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU) after the resection of the tumor by real-time reverse transcription-polymerase chain reaction (RT-PCR) using TaqMan probe. 2. The mean absolute value of MGMTmRNA normalized to the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for 50 tumors was 1.29 x 10(4)+/- 1.28 x 10(4) copy/microg RNA (mean +/- SD). The amount of MGMTmRNA less than 6 x 10(3) copy/microg RNA was the most significant factor in predicting the initial effect of treatment with ACNU by multi-variant regression analysis (p = 0.0157). 3. These results suggest that quantitation of MGMTmRNA is the excellent method for predicting for the effect of ACNU in glioma therapy.

Development and application of an oligonucleotide microarray and real-time quantitative PCR for detection of wastewater bacterial pathogens

Energy Technology Data Exchange (ETDEWEB)

Lee, Dae-Young [National Water Research Institute, Environment Canada, 867 Lakeshore Road, Burlington, Ontario, L7R 4A6 (Canada)], E-mail: daeyoung.lee@ec.gc.ca; Lauder, Heather; Cruwys, Heather; Falletta, Patricia [National Water Research Institute, Environment Canada, 867 Lakeshore Road, Burlington, Ontario, L7R 4A6 (Canada); Beaudette, Lee A. [Environmental Science and Technology Centre, Environment Canada, 335 River Road South, Ottawa, Ontario, K1A 0H3 (Canada)], E-mail: lee.beaudette@ec.gc.ca

2008-07-15

Conventional microbial water quality test methods are well known for their technical limitations, such as lack of direct pathogen detection capacity and low throughput capability. The microarray assay has recently emerged as a promising alternative for environmental pathogen monitoring. In this study, bacterial pathogens were detected in municipal wastewater using a microarray equipped with short oligonucleotide probes targeting 16S rRNA sequences. To date, 62 probes have been designed against 38 species, 4 genera, and 1 family of pathogens. The detection sensitivity of the microarray for a waterborne pathogen Aeromonas hydrophila was determined to be approximately 1.0% of the total DNA, or approximately 10{sup 3}A. hydrophila cells per sample. The efficacy of the DNA microarray was verified in a parallel study where pathogen genes and E. coli cells were enumerated using real-time quantitative PCR (qPCR) and standard membrane filter techniques, respectively. The microarray and qPCR successfully detected multiple wastewater pathogen species at different stages of the disinfection process (i.e. secondary effluents vs. disinfected final effluents) and at two treatment plants employing different disinfection methods (i.e. chlorination vs. UV irradiation). This result demonstrates the effectiveness of the DNA microarray as a semi-quantitative, high throughput pathogen monitoring tool for municipal wastewater.

Recombinant plasmid-based quantitative Real-Time PCR analysis of Salmonella enterica serotypes and its application to milk samples.

Science.gov (United States)

Gokduman, Kurtulus; Avsaroglu, M Dilek; Cakiris, Aris; Ustek, Duran; Gurakan, G Candan

2016-03-01

The aim of the current study was to develop, a new, rapid, sensitive and quantitative Salmonella detection method using a Real-Time PCR technique based on an inexpensive, easy to produce, convenient and standardized recombinant plasmid positive control. To achieve this, two recombinant plasmids were constructed as reference molecules by cloning the two most commonly used Salmonella-specific target gene regions, invA and ttrRSBC. The more rapid detection enabled by the developed method (21 h) compared to the traditional culture method (90 h) allows the quantitative evaluation of Salmonella (quantification limits of 10(1)CFU/ml and 10(0)CFU/ml for the invA target and the ttrRSBC target, respectively), as illustrated using milk samples. Three advantages illustrated by the current study demonstrate the potential of the newly developed method to be used in routine analyses in the medical, veterinary, food and water/environmental sectors: I--The method provides fast analyses including the simultaneous detection and determination of correct pathogen counts; II--The method is applicable to challenging samples, such as milk; III--The method's positive controls (recombinant plasmids) are reproducible in large quantities without the need to construct new calibration curves. Copyright © 2016 Elsevier B.V. All rights reserved.

Spatial filtering velocimetry for real-time out-of-plane displacement measurements

DEFF Research Database (Denmark)

Olesen, Anders Sig; Yura, H.T.; Jakobsen, Michael Linde

2016-01-01

power spectrum of the photocurrent produced by this filter. This main contribution of this paper is a model, which describe the selectivity of the sensor, applied to speckle dynamics generated by an object moving out-of-plane. To motivate our interest in these filters we also present an all optical......We probe the dynamics of objective laser speckles as the axial distance between the object and the observation plane changes. With the purpose of measuring out-of-plane motion in real time, we apply optical spatial filtering velocimetry to the speckle dynamics. To achieve this, a rotationally...... symmetric spatial filter is designed. The spatial filter converts the speckle dynamics into a photocurrent with a quasi-sinusoidal response to the out-of-plane motion. The selectivity of the sensor relates directly to the uncertainty on sensor measurements. The selectivity most be derived from a temporal...

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.

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.

WetLab-2: Tools for Conducting On-Orbit Quantitative Real-Time Gene Expression Analysis on ISS

Science.gov (United States)

Parra, Macarena; Almeida, Eduardo; Boone, Travis; Jung, Jimmy; Schonfeld, Julie

2014-01-01

The objective of NASA Ames Research Centers WetLab-2 Project is to place on the ISS a research platform capable of conducting gene expression analysis via quantitative real-time PCR (qRT-PCR) of biological specimens sampled or cultured on orbit. The project has selected a Commercial-Off-The-Shelf (COTS) qRT-PCR system, the Cepheid SmartCycler and will fly it in its COTS configuration. The SmartCycler has a number of advantages including modular design (16 independent PCR modules), low power consumption, rapid ramp times and the ability to detect up to four separate fluorescent channels at one time enabling multiplex assays that can be used for normalization and to study multiple genes of interest in each module. The team is currently working with Cepheid to enable the downlink of data from the ISS to the ground and provide uplink capabilities for programming, commanding, monitoring, and instrument maintenance. The project has adapted commercial technology to design a module that can lyse cells and extract RNA of sufficient quality and quantity for use in qRT-PCR reactions while using a housekeeping gene to normalize RNA concentration and integrity. The WetLab-2 system is capable of processing multiple sample types ranging from microbial cultures to animal tissues dissected on-orbit. The ability to conduct qRT-PCR on-orbit eliminates the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of fixed samples. The system can be used to validate terrestrial analyses of samples returned from ISS by providing on-orbit gene expression benchmarking prior to sample return. The ability to get on orbit data will provide investigators with the opportunity to adjust experiment parameters for subsequent trials based on the real-time data analysis without need for sample return and re-flight. Researchers will also be able to sample multigenerational changes in organisms. Finally, the system can be

Evaluation of real-time operating system for small-scale embedded systems

International Nuclear Information System (INIS)

Dayang Norhayati Abang Jawawi; Rosbi Mamat

1999-01-01

In this paper, the performance of some real-time operating systems for small-scale embedded systems are evaluated based on some criteria. The evaluation is performed qualitatively and quantitatively. The evaluation results based on a case study on an engineering application will be presented. (author)

Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR.

Science.gov (United States)

Mwaengo, D; Lorenzo, G; Iglesias, J; Warigia, M; Sang, R; Bishop, R P; Brun, A

2012-10-01

Diagnostic methods allowing for rapid identification of pathogens are crucial for controlling and preventing dissemination after disease outbreaks as well as for use in surveillance programs. For arboviruses, detection of the presence of virus in their arthropod hosts is important for monitoring of viral activity and quantitative information is useful for modeling of transmission dynamics. In this study, molecular detection of Rift Valley fever virus (RVFV) in mosquito samples from the 2006 to 2007 East African outbreaks was performed using quantitative real-time PCR assay (qRT-PCR). Specific RVFV sequence-based primer/fluorogenic (TaqMan) probe sets were derived from the L and S RNA segments of the virus. Both primer-probe L and S segment-based combinations detected genomic RVFV sequences, with generally comparable levels of sensitivity. Viral loads from three mosquito species, Aedes mcintoshi, Aedes ochraceus and Mansonia uniformis were estimated and significant differences of between 5- and 1000-fold were detected between Ae. mcintoshi and M. uniformis using both the L and S primer-probe-based assays. The genetic relationships of the viral sequences in mosquito samples were established by partial M segment sequencing and assigned to the two previously described viral lineages defined by analysis of livestock isolates obtained during the 2006-2007 outbreak, confirming that similar viruses were present in both the vector and mammalian host. The data confirms the utility of qRT-PCR for identification and initial quantification of virus in mosquito samples during RVFV outbreaks. Copyright © 2012 Elsevier B.V. All rights reserved.

MAC-Level Communication Time Modeling and Analysis for Real-Time WSNs

Directory of Open Access Journals (Sweden)

STANGACIU, V.

2016-02-01

Full Text Available Low-level communication protocols and their timing behavior are essential to developing wireless sensor networks (WSNs able to provide the support and operating guarantees required by many current real-time applications. Nevertheless, this aspect still remains an issue in the state-of-the-art. In this paper we provide a detailed analysis of a recently proposed MAC-level communication timing model and demonstrate its usability in designing real-time protocols. The results of a large set of measurements are also presented and discussed here, in direct relation to the main time parameters of the analyzed model.

Hardware Approach for Real Time Machine Stereo Vision

Directory of Open Access Journals (Sweden)

Michael Tornow

2006-02-01

Full Text Available Image processing is an effective tool for the analysis of optical sensor information for driver assistance systems and controlling of autonomous robots. Algorithms for image processing are often very complex and costly in terms of computation. In robotics and driver assistance systems, real-time processing is necessary. Signal processing algorithms must often be drastically modified so they can be implemented in the hardware. This task is especially difficult for continuous real-time processing at high speeds. This article describes a hardware-software co-design for a multi-object position sensor based on a stereophotogrammetric measuring method. In order to cover a large measuring area, an optimized algorithm based on an image pyramid is implemented in an FPGA as a parallel hardware solution for depth map calculation. Object recognition and tracking are then executed in real-time in a processor with help of software. For this task a statistical cluster method is used. Stabilization of the tracking is realized through use of a Kalman filter. Keywords: stereophotogrammetry, hardware-software co-design, FPGA, 3-d image analysis, real-time, clustering and tracking.

Research on Control Method Based on Real-Time Operational Reliability Evaluation for Space Manipulator

Directory of Open Access Journals (Sweden)

Yifan Wang

2014-05-01

Full Text Available A control method based on real-time operational reliability evaluation for space manipulator is presented for improving the success rate of a manipulator during the execution of a task. In this paper, a method for quantitative analysis of operational reliability is given when manipulator is executing a specified task; then a control model which could control the quantitative operational reliability is built. First, the control process is described by using a state space equation. Second, process parameters are estimated in real time using Bayesian method. Third, the expression of the system's real-time operational reliability is deduced based on the state space equation and process parameters which are estimated using Bayesian method. Finally, a control variable regulation strategy which considers the cost of control is given based on the Theory of Statistical Process Control. It is shown via simulations that this method effectively improves the operational reliability of space manipulator control system.

[Detecting HB-1 Expression Level in Bone Marrow of Acute Leukemia Patients by Real-Time Fluorescence Quantitative RT-PCR].

Science.gov (United States)

Wang, Qing-Yun; Li, Yuan; Ji, Li; Liang, Ze-Yin; Liu, Wei; Ren, Han-Yun; Qiu, Zhi-Xiang

2018-02-01

To investigate the expression level of HB-1 gene in patients with acute lymphoblastic leukemia (ALL) and the significance of HB-1 gene in monitoring of minimal residual disease (MRD). The method of real-time fluorescence quantitative RT-PCR (Taqman probe) was established to detect the expression levels of HB-1 gene; then the sensitivity, specificity and repeatability of this assay were evaluated and verified. The HB-1 gene expression levels in bone marrow of 183 cases of ALL, 70 cases of acute myeloid leukemias (AML), 52 cases of non-malignant hematologic diseases and 24 healthy hematopoietic stem cell donors were detected. The correlation of HB-1 level with diagnosis and relapse was analyzed by detecting bone marrow samples of 33 B-ALL. The sensitivity of this assay reached the 10 -4 level. The coefficient of variation for inter-batch and inter-tube of HB-1 were 6.79% and 4.80%, respectively. It was found that HB-1 gene specifically expressed in acute B lymphoblastic leukemia. The median expression levels of HB-1 gene in newly diagnosed and relapsed B-ALL patients were statistically significantly higher than those in ALL in complete remission(CR), newly diagnosed T-ALL, newly diagnosed AML, non-malignant hematologic diseases, and healthy hematopoietic stem cell donors(33.0% vs 0.68%, 0.07%, 0.02%, 0.58% and 0, respectively) (P0.05). The expression level of HB-1 gene declined sharply when B-ALL patients reached complete remission (0-7.99%, with median level 0.68%), but increased when relapsed (7.69%, 8.08% and 484.0% in 3 relapsed samples), which was in accordance with results of flow cytometry. HB-1 gene specifically expressed in acute B lymphoblastic leukemia cells. The established real-time fluorescence quantitative RT-PCR assay shows good sensitivity, specificity and repeatability, thus, can be used as a biological marker in the clinical detection, monitoring MRD and predicting of early relapse for B-ALL patients.

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.

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.

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.

Quasi-real-time photon pulse duration measurement by analysis of FEL radiation spectra

Energy Technology Data Exchange (ETDEWEB)

Engel, Robin, E-mail: robin.engel@uni-oldenburg.de [Deutsches Elektronen-Synchrotron, Notkestrasse 85, D-22603 Hamburg (Germany); Institut für Physik, Carl von Ossietzky Universität Oldenburg, D-26111 Oldenburg (Germany); Institut für Laser und Optik, Hochschule Emden/Leer, University of Applied Sciences, Constantiaplatz 4, D-26723 Emden (Germany); Düsterer, Stefan; Brenner, Günter [Deutsches Elektronen-Synchrotron, Notkestrasse 85, D-22603 Hamburg (Germany); Teubner, Ulrich [Deutsches Elektronen-Synchrotron, Notkestrasse 85, D-22603 Hamburg (Germany); Institut für Physik, Carl von Ossietzky Universität Oldenburg, D-26111 Oldenburg (Germany); Institut für Laser und Optik, Hochschule Emden/Leer, University of Applied Sciences, Constantiaplatz 4, D-26723 Emden (Germany)

2016-01-01

Considering the second-order spectral correlation function of SASE-FEL radiation allows a real-time observation of the photon pulse duration during spectra acquisition. For photon diagnostics at free-electron lasers (FELs), the determination of the photon pulse duration is an important challenge and a complex task. This is especially true for SASE FELs with strongly fluctuating pulse parameters. However, most techniques require an extensive experimental setup, data acquisition and evaluation time, limiting the usability in all-day operation. In contrast, the presented work uses an existing approach based on the analysis of statistical properties of measured SASE FEL spectra and implements it as a software tool, integrated in FLASH’s data acquisition system. This allows the calculation of the average pulse durations from a set of measured spectral distributions with only seconds of delay, whenever high-resolution spectra are recorded.

Development of a real-time RT-PCR assay for the simultaneous identification, quantitation and differentiation of avian metapneumovirus subtypes A and B.

Science.gov (United States)

Cecchinato, Mattia; Lupini, Caterina; Munoz Pogoreltseva, Olga Svetlana; Listorti, Valeria; Mondin, Alessandra; Drigo, Michele; Catelli, Elena

2013-01-01

In recent years, special attention has been paid to real-time polymerase chain reaction (PCR) for avian metapneumovirus (AMPV) diagnosis, due to its numerous advantages over classical PCR. A new multiplex quantitative real-time reverse transcription-PCR (qRT-PCR) with molecular beacon probe assay, designed to target the SH gene, was developed. The test was evaluated in terms of specificity, sensitivity and repeatability, and compared with conventional RT nested-PCR based on the G gene. All of the AMPV subtype A and B strains tested were amplified and specifically detected while no amplification occurred with other non-target bird respiratory pathogens. The detection limit of the assay was 10(-0.41) median infectious dose/ml and 10(1.15) median infectious dose/ml when the AMPV-B strain IT/Ty/B/Vr240/87 and the AMPV-A strain IT/Ty/A/259-01/03 were used, respectively, as templates. In all cases, the amplification efficiency was approximately 2 and the error values were 0.9375) between crossing point values and virus quantities, making the assay herein designed reliable for quantification. When the newly developed qRT-PCR was compared with a conventional RT nested-PCR, it showed greater sensitivity with RNA extracted from both positive controls and from experimentally infected birds. This assay can be effectively used for the detection, identification, differentiation and quantitation of AMPV subtype A or subtype B to assist in disease diagnosis and to carry out rapid surveillance with high levels of sensitivity and specificity.

Gamma-ray imaging system for real-time measurements in nuclear waste characterisation

Science.gov (United States)

Caballero, L.; Albiol Colomer, F.; Corbi Bellot, A.; Domingo-Pardo, C.; Leganés Nieto, J. L.; Agramunt Ros, J.; Contreras, P.; Monserrate, M.; Olleros Rodríguez, P.; Pérez Magán, D. L.

2018-03-01

A compact, portable and large field-of-view gamma camera that is able to identify, locate and quantify gamma-ray emitting radioisotopes in real-time has been developed. The device delivers spectroscopic and imaging capabilities that enable its use it in a variety of nuclear waste characterisation scenarios, such as radioactivity monitoring in nuclear power plants and more specifically for the decommissioning of nuclear facilities. The technical development of this apparatus and some examples of its application in field measurements are reported in this article. The performance of the presented gamma-camera is also benchmarked against other conventional techniques.

Plasmon-induced photoelectrochemical biosensor for in situ real-time measurement of biotin-streptavidin binding kinetics under visible light irradiation

International Nuclear Information System (INIS)

Guo, Jingchun; Oshikiri, Tomoya; Ueno, Kosei; Shi, Xu; Misawa, Hiroaki

2017-01-01

We developed a localized surface plasmon-induced visible light-responsive photoelectrochemical (PEC) biosensor using a titanium dioxide (TiO_2) photoelectrode loaded with gold nanoislands (AuNIs) for in situ real-time measurement of biotin-streptavidin association. As a proof of concept, self-assembled thiol-terminated biotin molecules bound on a AuNIs/TiO_2 photoelectrode were successfully utilized to explore the photocurrent response to streptavidin-modified gold nanoparticle (STA-AuNP) solutions. This plasmon-induced PEC biosensor is simple and easy to miniaturize. Additionally, the PEC biosensor achieves highly sensitive measurements under only visible light irradiation and prevents the UV-induced damage of samples. Furthermore, a novel approach has been proposed to realize the real-time monitoring of biotin-STA binding affinities and kinetics by analyzing the PEC sensing characteristics. This PEC biosensor and novel analysis method could provide a new approach for the specific electrical detection and real-time kinetic measurements for clinical diagnostics and drug development. - Highlights: • A plasmon-induced visible light-responsive photoelectrochemical biosensor is developed and the system can be miniaturized.

Plasmon-induced photoelectrochemical biosensor for in situ real-time measurement of biotin-streptavidin binding kinetics under visible light irradiation

Energy Technology Data Exchange (ETDEWEB)

Guo, Jingchun; Oshikiri, Tomoya; Ueno, Kosei; Shi, Xu [Research Institute for Electronic Science, Hokkaido University, Sapporo 001-0021 (Japan); Misawa, Hiroaki, E-mail: misawa@es.hokudai.ac.jp [Research Institute for Electronic Science, Hokkaido University, Sapporo 001-0021 (Japan); Department of Applied Chemistry & Institute of Molecular Science, National Chiao Tung University, Hsinchu 30010, Taiwan (China)

2017-03-08

We developed a localized surface plasmon-induced visible light-responsive photoelectrochemical (PEC) biosensor using a titanium dioxide (TiO{sub 2}) photoelectrode loaded with gold nanoislands (AuNIs) for in situ real-time measurement of biotin-streptavidin association. As a proof of concept, self-assembled thiol-terminated biotin molecules bound on a AuNIs/TiO{sub 2} photoelectrode were successfully utilized to explore the photocurrent response to streptavidin-modified gold nanoparticle (STA-AuNP) solutions. This plasmon-induced PEC biosensor is simple and easy to miniaturize. Additionally, the PEC biosensor achieves highly sensitive measurements under only visible light irradiation and prevents the UV-induced damage of samples. Furthermore, a novel approach has been proposed to realize the real-time monitoring of biotin-STA binding affinities and kinetics by analyzing the PEC sensing characteristics. This PEC biosensor and novel analysis method could provide a new approach for the specific electrical detection and real-time kinetic measurements for clinical diagnostics and drug development. - Highlights: • A plasmon-induced visible light-responsive photoelectrochemical biosensor is developed and the system can be miniaturized.

A novel universal real-time PCR system using the attached universal duplex probes for quantitative analysis of nucleic acids

Directory of Open Access Journals (Sweden)

Wilson Zoe A

2008-06-01

Full Text Available Abstract Background Real-time PCR techniques are being widely used for nucleic acids analysis, but one limitation of current frequently employed real-time PCR is the high cost of the labeled probe for each target molecule. Results We describe a real-time PCR technique employing attached universal duplex probes (AUDP, which has the advantage of generating fluorescence by probe hydrolysis and strand displacement over current real-time PCR methods. AUDP involves one set of universal duplex probes in which the 5' end of the fluorescent probe (FP and a complementary quenching probe (QP lie in close proximity so that fluorescence can be quenched. The PCR primer pair with attached universal template (UT and the FP are identical to the UT sequence. We have shown that the AUDP technique can be used for detecting multiple target DNA sequences in both simplex and duplex real-time PCR assays for gene expression analysis, genotype identification, and genetically modified organism (GMO quantification with comparable sensitivity, reproducibility, and repeatability with other real-time PCR methods. Conclusion The results from GMO quantification, gene expression analysis, genotype identification, and GMO quantification using AUDP real-time PCR assays indicate that the AUDP real-time PCR technique has been successfully applied in nucleic acids analysis, and the developed AUDP real-time PCR technique will offer an alternative way for nucleic acid analysis with high efficiency, reliability, and flexibility at low cost.

Real-time TaqMan polymerase chain reaction to quantify the effects ...

African Journals Online (AJOL)

TaqMan polymerase chain reaction was developed to quantify the number of Bifidobacterium. We used this assay to detect genomic DNA of Bifidobacterium in the intestinal tract digesta of piglets, including duodenum, jejunum, ileum, cecum and colon. Our results indicated that, developed new real-time quantitative PCR ...

Microgravity validation of a novel system for RNA isolation and multiplex quantitative real time PCR analysis of gene expression on the International Space Station.

Directory of Open Access Journals (Sweden)

Macarena Parra

Full Text Available The International Space Station (ISS National Laboratory is dedicated to studying the effects of space on life and physical systems, and to developing new science and technologies for space exploration. A key aspect of achieving these goals is to operate the ISS National Lab more like an Earth-based laboratory, conducting complex end-to-end experimentation, not limited to simple microgravity exposure. Towards that end NASA developed a novel suite of molecular biology laboratory tools, reagents, and methods, named WetLab-2, uniquely designed to operate in microgravity, and to process biological samples for real-time gene expression analysis on-orbit. This includes a novel fluidic RNA Sample Preparation Module and fluid transfer devices, all-in-one lyophilized PCR assays, centrifuge, and a real-time PCR thermal cycler. Here we describe the results from the WetLab-2 validation experiments conducted in microgravity during ISS increment 47/SPX-8. Specifically, quantitative PCR was performed on a concentration series of DNA calibration standards, and Reverse Transcriptase-quantitative PCR was conducted on RNA extracted and purified on-orbit from frozen Escherichia coli and mouse liver tissue. Cycle threshold (Ct values and PCR efficiencies obtained on-orbit from DNA standards were similar to Earth (1 g controls. Also, on-orbit multiplex analysis of gene expression from bacterial cells and mammalian tissue RNA samples was successfully conducted in about 3 h, with data transmitted within 2 h of experiment completion. Thermal cycling in microgravity resulted in the trapping of gas bubbles inside septa cap assay tubes, causing small but measurable increases in Ct curve noise and variability. Bubble formation was successfully suppressed in a rapid follow-up on-orbit experiment using standard caps to pressurize PCR tubes and reduce gas release during heating cycles. The WetLab-2 facility now provides a novel operational on-orbit research capability for

Development of a real-time quantitative assay for detection of Epstein-Barr virus

NARCIS (Netherlands)

H.G.M. Niesters (Bert); E. Fries; K.C. Wolthers (Katja); A.D.M.E. Osterhaus (Albert); J.J. Cornelissen (Jan); J.W.J. van Esser (Joost)

2000-01-01

textabstractWith the use of real-time PCR, we developed and evaluated a rapid, sensitive, specific, and reproducible method for the detection of Epstein-Barr virus (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and

Real-Time Forecasting Revisited: Letting the Data Decide

OpenAIRE

Jackson Kitchen; John Kitchen

2013-01-01

Real-time GDP forecasting, also often known as “nowcasting,” produces estimates for current-quarter real GDP growth, typically based on a centered value from a set of estimates from incoming indicators. These real-time measures are usually intended to be data-based and to not be based on forecaster judgment or add factors. Even so, estimation methodologies in this research area—and prior versions of the system we use—typically have been constrained by using various “fixed” relationships, such...

Real-time x-ray scattering study of the initial growth of organic crystals on polymer brushes

Energy Technology Data Exchange (ETDEWEB)

An, Sung Yup; Ahn, Kwangseok; Kim, Doris Yangsoo; Lee, Dong Ryeol, E-mail: drlee@ssu.ac.kr [Department of Physics, Soongsil University, Seoul 156-743 (Korea, Republic of); Lee, Hyun-Hwi [Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang 790-784 (Korea, Republic of); Cho, Jeong Ho, E-mail: jhcho94@skku.edu [Department of Chemical Engineering, SKKU Advanced Institute of Nanotechnology (SAINT) and Center for Human Interface Nano Technology (HINT), Sungkyunkwan University, Suwon 440-476 (Korea, Republic of)

2014-04-21

We studied the early-stage growth structures of pentacene organic crystals grown on polymer brushes using real-time x-ray scattering techniques. In situ x-ray reflectivity and atomic force microscopy analyses revealed that at temperatures close to the glass transition temperature of polymer brush, the pentacene overlayer on a polymer brush film showed incomplete condensation and 3D island structures from the first monolayer. A growth model based on these observations was used to quantitatively analyze the real-time anti-Bragg x-ray scattering intensities measured during pentacene growth to obtain the time-dependent layer coverage of the individual pentacene monolayers. The extracted total coverage confirmed significant desorption and incomplete condensation in the pentacene films deposited on the polymer brushes. These effects are ascribed to the change in the surface viscoelasticity of the polymer brushes around the glass transition temperature.

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

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

Novel Quantitative Real-Time LCR for the Sensitive Detection of SNP Frequencies in Pooled DNA: Method Development, Evaluation and Application

Science.gov (United States)

Psifidi, Androniki; Dovas, Chrysostomos; Banos, Georgios

2011-01-01

Background Single nucleotide polymorphisms (SNP) have proven to be powerful genetic markers for genetic applications in medicine, life science and agriculture. A variety of methods exist for SNP detection but few can quantify SNP frequencies when the mutated DNA molecules correspond to a small fraction of the wild-type DNA. Furthermore, there is no generally accepted gold standard for SNP quantification, and, in general, currently applied methods give inconsistent results in selected cohorts. In the present study we sought to develop a novel method for accurate detection and quantification of SNP in DNA pooled samples. Methods The development and evaluation of a novel Ligase Chain Reaction (LCR) protocol that uses a DNA-specific fluorescent dye to allow quantitative real-time analysis is described. Different reaction components and thermocycling parameters affecting the efficiency and specificity of LCR were examined. Several protocols, including gap-LCR modifications, were evaluated using plasmid standard and genomic DNA pools. A protocol of choice was identified and applied for the quantification of a polymorphism at codon 136 of the ovine PRNP gene that is associated with susceptibility to a transmissible spongiform encephalopathy in sheep. Conclusions The real-time LCR protocol developed in the present study showed high sensitivity, accuracy, reproducibility and a wide dynamic range of SNP quantification in different DNA pools. The limits of detection and quantification of SNP frequencies were 0.085% and 0.35%, respectively. Significance The proposed real-time LCR protocol is applicable when sensitive detection and accurate quantification of low copy number mutations in DNA pools is needed. Examples include oncogenes and tumour suppressor genes, infectious diseases, pathogenic bacteria, fungal species, viral mutants, drug resistance resulting from point mutations, and genetically modified organisms in food. PMID:21283808

Robust real-time extraction of respiratory signals from PET list-mode data.

Science.gov (United States)

Salomon, Andre; Zhang, Bin; Olivier, Patrick; Goedicke, Andreas

2018-05-01

Respiratory motion, which typically cannot simply be suspended during PET image acquisition, affects lesions' detection and quantitative accuracy inside or in close vicinity to the lungs. Some motion compensation techniques address this issue via pre-sorting ("binning") of the acquired PET data into a set of temporal gates, where each gate is assumed to be minimally affected by respiratory motion. Tracking respiratory motion is typically realized using dedicated hardware (e.g. using respiratory belts and digital cameras). Extracting respiratory signalsdirectly from the acquired PET data simplifies the clinical workflow as it avoids to handle additional signal measurement equipment. We introduce a new data-driven method "Combined Local Motion Detection" (CLMD). It uses the Time-of-Flight (TOF) information provided by state-of-the-art PET scanners in order to enable real-time respiratory signal extraction without additional hardware resources. CLMD applies center-of-mass detection in overlapping regions based on simple back-positioned TOF event sets acquired in short time frames. Following a signal filtering and quality-based pre-selection step, the remaining extracted individual position information over time is then combined to generate a global respiratory signal. The method is evaluated using 7 measured FDG studies from single and multiple scan positions of the thorax region, and it is compared to other software-based methods regarding quantitative accuracy and statistical noise stability. Correlation coefficients around 90% between the reference and the extracted signal have been found for those PET scans where motion affected features such as tumors or hot regions were present in the PET field-of-view. For PET scans with a quarter of typically applied radiotracer doses, the CLMD method still provides similar high correlation coefficients which indicates its robustness to noise. Each CLMD processing needed less than 0.4s in total on a standard multi-core CPU

Robust real-time extraction of respiratory signals from PET list-mode data

Science.gov (United States)

Salomon, André; Zhang, Bin; Olivier, Patrick; Goedicke, Andreas

2018-06-01

Respiratory motion, which typically cannot simply be suspended during PET image acquisition, affects lesions’ detection and quantitative accuracy inside or in close vicinity to the lungs. Some motion compensation techniques address this issue via pre-sorting (‘binning’) of the acquired PET data into a set of temporal gates, where each gate is assumed to be minimally affected by respiratory motion. Tracking respiratory motion is typically realized using dedicated hardware (e.g. using respiratory belts and digital cameras). Extracting respiratory signals directly from the acquired PET data simplifies the clinical workflow as it avoids handling additional signal measurement equipment. We introduce a new data-driven method ‘combined local motion detection’ (CLMD). It uses the time-of-flight (TOF) information provided by state-of-the-art PET scanners in order to enable real-time respiratory signal extraction without additional hardware resources. CLMD applies center-of-mass detection in overlapping regions based on simple back-positioned TOF event sets acquired in short time frames. Following a signal filtering and quality-based pre-selection step, the remaining extracted individual position information over time is then combined to generate a global respiratory signal. The method is evaluated using seven measured FDG studies from single and multiple scan positions of the thorax region, and it is compared to other software-based methods regarding quantitative accuracy and statistical noise stability. Correlation coefficients around 90% between the reference and the extracted signal have been found for those PET scans where motion affected features such as tumors or hot regions were present in the PET field-of-view. For PET scans with a quarter of typically applied radiotracer doses, the CLMD method still provides similar high correlation coefficients which indicates its robustness to noise. Each CLMD processing needed less than 0.4 s in total on a standard

Ex vivo screening for immunodominant viral epitopes by quantitative real time polymerase chain reaction (qRT-PCR

Directory of Open Access Journals (Sweden)

Nagorsen Dirk

2003-12-01

Full Text Available Abstract The identification and characterization of viral epitopes across the Human Leukocyte Antigen (HLA polymorphism is critical for the development of actives-specific or adoptive immunotherapy of virally-mediated diseases. This work investigates whether cytokine mRNA transcripts could be used to identify epitope-specific HLA-restricted memory T lymphocytes reactivity directly in fresh peripheral blood mononuclear cells (PBMCs from viral-seropositive individuals in response to ex vivo antigen recall. PBMCs from HLA-A*0201 healthy donors, seropositive for Cytomegalovirus (CMV and Influenza (Flu, were exposed for different periods and at different cell concentrations to the HLA-A*0201-restricted viral FluM158–66 and CMVpp65495–503 peptides. Quantitative real time PCR (qRT-PCR was employed to evaluate memory T lymphocyte immune reactivation by measuring the production of mRNA encoding four cytokines: Interferon-γ (IFN-γ, Interleukin-2 (IL-2, Interleukin-4 (IL-4, and Interleukin-10 (IL-10. We could characterize cytokine expression kinetics that illustrated how cytokine mRNA levels could be used as ex vivo indicators of T cell reactivity. Particularly, IFN-γ mRNA transcripts could be consistently detected within 3 to 12 hours of short-term stimulation in levels sufficient to screen for HLA-restricted viral immune responses in seropositive subjects. This strategy will enhance the efficiency of the identification of viral epitopes independently of the individual HLA phenotype and could be used to follow the intensity of immune responses during disease progression or in response to in vivo antigen-specific immunization.

Ex vivo screening for immunodominant viral epitopes by quantitative real time polymerase chain reaction (qRT-PCR)

Science.gov (United States)

Provenzano, Maurizio; Mocellin, Simone; Bonginelli, Paola; Nagorsen, Dirk; Kwon, Seog-Woon; Stroncek, David

2003-01-01

The identification and characterization of viral epitopes across the Human Leukocyte Antigen (HLA) polymorphism is critical for the development of actives-specific or adoptive immunotherapy of virally-mediated diseases. This work investigates whether cytokine mRNA transcripts could be used to identify epitope-specific HLA-restricted memory T lymphocytes reactivity directly in fresh peripheral blood mononuclear cells (PBMCs) from viral-seropositive individuals in response to ex vivo antigen recall. PBMCs from HLA-A*0201 healthy donors, seropositive for Cytomegalovirus (CMV) and Influenza (Flu), were exposed for different periods and at different cell concentrations to the HLA-A*0201-restricted viral FluM158–66 and CMVpp65495–503 peptides. Quantitative real time PCR (qRT-PCR) was employed to evaluate memory T lymphocyte immune reactivation by measuring the production of mRNA encoding four cytokines: Interferon-γ (IFN-γ), Interleukin-2 (IL-2), Interleukin-4 (IL-4), and Interleukin-10 (IL-10). We could characterize cytokine expression kinetics that illustrated how cytokine mRNA levels could be used as ex vivo indicators of T cell reactivity. Particularly, IFN-γ mRNA transcripts could be consistently detected within 3 to 12 hours of short-term stimulation in levels sufficient to screen for HLA-restricted viral immune responses in seropositive subjects. This strategy will enhance the efficiency of the identification of viral epitopes independently of the individual HLA phenotype and could be used to follow the intensity of immune responses during disease progression or in response to in vivo antigen-specific immunization. PMID:14675481

Development of a system for real-time measurements of metabolite transport in plants using short-lived positron-emitting radiotracers

Science.gov (United States)

Kiser, Matthew R.

that enabled the quantitative biological studies reported in this thesis were developed through a series of experiments made at TUNL and the Phytotron. Collimated single detectors and coincidence counting techniques were used to monitor the radiotracer distribution on a coarse spatial scale. Additionally, a prototype Versatile Imager for Positron Emitting Radiotracers (VIPER) was built to provide the capability of measuring radiotracer distributions in plants with high spatial resolution (˜2.5 mm). This device enables detailed quantification of real-time metabolite dynamics on fine spatial scales. The full capabilities of this radiotracer system were utilized in an investigation of the effects of elevated atmospheric CO2 concentration and root nutrient availability on the transport and allocation of recently fixed carbon, including that released from the roots via exudation or respiration, in two grass species. The 11CO2 gas was introduced to a leaf on the plants grown at either ambient or elevated atmospheric CO 2. Two sequential measurements were performed per day on each plant: a control nutrient solution labeling immediately followed by labeling with a 10-fold increase or decrease in nutrient concentration. The real-time distribution of 11C-labeled photoassimilate was measured in vivo throughout the plant and root environment. This measurement resulted in the first observation of a rapid plant response to short-term changes in nutrient availability via correlated changes in the photoassimilate allocation to root exudates. Our data indicated that root exudation was consistently enhanced at lower nutrient concentrations. Also, we found that elevated atmospheric CO2 increased the velocity of photoassimilate transport throughout the plant, enhanced root exudation in an annual crop grass, and reduced root exudation in a perennial native grass.

Correlation between electrochemical impedance measurements and corrosion rate of magnesium investigated by real-time hydrogen measurement and optical imaging

OpenAIRE

Curioni, M.; Scenini, F.; Monetta, T.; Bellucci, F.

2015-01-01

The corrosion behaviour of magnesium in chloride-containing aqueous environment was investigated by potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) performed simultaneously with real-time hydrogen evolution measurements and optical imaging of the corroding surface. The potentiodynamic investigation revealed substantial deviations from linearity in close proximity of the corrosion potential. In particular, differences in the slope of the current/potential curves w...

Estimating marginal properties of quantitative real-time PCR data using nonlinear mixed models

DEFF Research Database (Denmark)

Gerhard, Daniel; Bremer, Melanie; Ritz, Christian

2014-01-01

A unified modeling framework based on a set of nonlinear mixed models is proposed for flexible modeling of gene expression in real-time PCR experiments. Focus is on estimating the marginal or population-based derived parameters: cycle thresholds and ΔΔc(t), but retaining the conditional mixed mod...

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

Real Time Management of the AD Schottky/BTF Beam Measurement

CERN Document Server

Angoletta, Maria Elena

2003-01-01

The AD Schottky and BTF system relies on rapid acquisition and analysis of beam quantisation noise during the AD cycle which is based on an embedded receiver and digital signal processing board hosted in a VME system. The software running in the VME sets up the embedded system and amplifiers, interfaces to the RF and control system, manages the execution speed and sequence constraints with respect to the various operating modes, schedules measurements during the AD cycle and performs post processing taking into account the beam conditions in an autonomous way. The operating modes of the instrument dynamically depend on a detailed configuration, the beam parameters during the AD cycle and optional user interaction. Various subsets of the processed data are available on line and in quasi real time for beam intensity, momentum spread and several spectrum types, which form an important part of AD operation today.

Quantification of bacteria adherent to gastrointestinal mucosa by real-time PCR

NARCIS (Netherlands)

Huijsdens, Xander W.; Linskens, Ronald K.; Mak, Mariëtte; Meuwissen, Stephan G. M.; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

2002-01-01

The use of real-time quantitative PCR (5' nuclease PCR assay) as a tool to study the gastrointestinal microflora that adheres to the colonic mucosa was evaluated. We developed primers and probes based on the 16S ribosomal DNA gene sequences for the detection of Escherichia coli and Bacteroides

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.

Real-time LMR control parameter generation using advanced adaptive synthesis

International Nuclear Information System (INIS)

King, R.W.; Mott, J.E.

1990-01-01

The reactor ''delta T'', the difference between the average core inlet and outlet temperatures, for the liquid-sodium-cooled Experimental Breeder Reactor 2 is empirically synthesized in real time from, a multitude of examples of past reactor operation. The real-time empirical synthesis is based on reactor operation. The real-time empirical synthesis is based on system state analysis (SSA) technology embodied in software on the EBR 2 data acquisition computer. Before the real-time system is put into operation, a selection of reactor plant measurements is made which is predictable over long periods encompassing plant shutdowns, core reconfigurations, core load changes, and plant startups. A serial data link to a personal computer containing SSA software allows the rapid verification of the predictability of these plant measurements via graphical means. After the selection is made, the real-time synthesis provides a fault-tolerant estimate of the reactor delta T accurate to +/-1%. 5 refs., 7 figs

Improvements for real-time magnetic equilibrium reconstruction on ASDEX Upgrade

International Nuclear Information System (INIS)

Giannone, L.; Fischer, R.; McCarthy, P.J.; Odstrcil, T.; Zammuto, I.; Bock, A.; Conway, G.; Fuchs, J.C.; Gude, A.; Igochine, V.; Kallenbach, A.; Lackner, K.; Maraschek, M.; Rapson, C.; Ruan, Q.; Schuhbeck, K.H.; Suttrop, W.; Wenzel, L.

2015-01-01

Highlights: • Spline basis current functions with second-order linear regularisation. • Perturbations of magnetic probe measurements due to ferromagnetic tiles on the inner wall and from oscillations in the fast position coil current are corrected. • A constraint of the safety factor on the magnetic axis is introduced. Soft X-ray tomography is used to assess the quality of the real-time magnetic equilibrium reconstruction. • External loop voltage measurements and magnetic probe pairs inside and outside the vessel wall were used to measure the vacuum vessel wall resistivity. - Abstract: Real-time magnetic equilibria are needed for NTM stabilization and disruption avoidance experiments on ASDEX Upgrade. Five improvements to real-time magnetic equilibrium reconstruction on ASDEX Upgrade have been investigated. The aim is to include as many features of the offline magnetic equilibrium reconstruction code in the real-time equilibrium reconstruction code. Firstly, spline current density basis functions with regularization are used in the offline equilibrium reconstruction code, CLISTE [1]. It is now possible to have the same number of spline basis functions in the real-time code. Secondly, in the presence of edge localized modes, (ELM's), it is found to be necessary to include the low pass filter effect of the vacuum vessel on the fast position control coil currents to correctly compensate the magnetic probes for current oscillations in these coils. Thirdly, the introduction of ferromagnetic tiles in ASDEX Upgrade means that a real-time algorithm for including the perturbations of the magnetic equilibrium generated by these tiles is required. A methodology based on tile surface currents is described. Fourthly, during current ramps it was seen that the difference between fitted and measured magnetic measurements in the equilibrium reconstruction were larger than in the constant current phase. External loop voltage measurements and magnetic probe pairs inside and

Improvements for real-time magnetic equilibrium reconstruction on ASDEX Upgrade

Energy Technology Data Exchange (ETDEWEB)

Giannone, L.; Fischer, R. [Max Planck Institute for Plasma Physics, 85748 Garching (Germany); McCarthy, P.J. [Department of Physics, University College Cork, Cork (Ireland); Odstrcil, T.; Zammuto, I.; Bock, A.; Conway, G.; Fuchs, J.C.; Gude, A.; Igochine, V.; Kallenbach, A.; Lackner, K.; Maraschek, M.; Rapson, C. [Max Planck Institute for Plasma Physics, 85748 Garching (Germany); Ruan, Q. [National Instruments, Austin, TX 78759-3504 (United States); Schuhbeck, K.H.; Suttrop, W. [Max Planck Institute for Plasma Physics, 85748 Garching (Germany); Wenzel, L. [National Instruments, Austin, TX 78759-3504 (United States)

2015-11-15

Highlights: • Spline basis current functions with second-order linear regularisation. • Perturbations of magnetic probe measurements due to ferromagnetic tiles on the inner wall and from oscillations in the fast position coil current are corrected. • A constraint of the safety factor on the magnetic axis is introduced. Soft X-ray tomography is used to assess the quality of the real-time magnetic equilibrium reconstruction. • External loop voltage measurements and magnetic probe pairs inside and outside the vessel wall were used to measure the vacuum vessel wall resistivity. - Abstract: Real-time magnetic equilibria are needed for NTM stabilization and disruption avoidance experiments on ASDEX Upgrade. Five improvements to real-time magnetic equilibrium reconstruction on ASDEX Upgrade have been investigated. The aim is to include as many features of the offline magnetic equilibrium reconstruction code in the real-time equilibrium reconstruction code. Firstly, spline current density basis functions with regularization are used in the offline equilibrium reconstruction code, CLISTE [1]. It is now possible to have the same number of spline basis functions in the real-time code. Secondly, in the presence of edge localized modes, (ELM's), it is found to be necessary to include the low pass filter effect of the vacuum vessel on the fast position control coil currents to correctly compensate the magnetic probes for current oscillations in these coils. Thirdly, the introduction of ferromagnetic tiles in ASDEX Upgrade means that a real-time algorithm for including the perturbations of the magnetic equilibrium generated by these tiles is required. A methodology based on tile surface currents is described. Fourthly, during current ramps it was seen that the difference between fitted and measured magnetic measurements in the equilibrium reconstruction were larger than in the constant current phase. External loop voltage measurements and magnetic probe pairs inside

Kurtosis based blind source extraction of complex noncircular signals with application in EEG artifact removal in real-time

Directory of Open Access Journals (Sweden)

Soroush eJavidi

2011-10-01

Full Text Available A new class of complex domain blind source extraction (BSE algorithms suitable for the extraction of both circular and noncircular complex signals is proposed. This is achieved through sequential extraction based on the degree of kurtosis and in the presence of noncircular measurement noise. The existence and uniqueness analysis of the solution is followed by a study of fast converging variants of the algorithm. The performance is first assessed through simulations on well understood benchmark signals, followed by a case study on real-time artifact removal from EEG signals, verified using both qualitative and quantitative metrics. The results illustrate the power of the proposed approach in real-time blind extraction of general complex-valued sources.

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.

Quantitative heartbeat coupling measures in human-horse interaction.

Science.gov (United States)

Lanata, Antonio; Guidi, Andrea; Valenza, Gaetano; Baragli, Paolo; Scilingo, Enzo Pasquale

2016-08-01

We present a study focused on a quantitative estimation of a human-horse dynamic interaction. A set of measures based on magnitude and phase coupling between heartbeat dynamics of both humans and horses in three different conditions is reported: no interaction, visual/olfactory interaction and grooming. Specifically, Magnitude Squared Coherence (MSC), Mean Phase Coherence (MPC) and Dynamic Time Warping (DTW) have been used as estimators of the amount of coupling between human and horse through the analysis of their heart rate variability (HRV) time series in a group of eleven human subjects, and one horse. The rationale behind this study is that the interaction of two complex biological systems go towards a coupling process whose dynamical evolution is modulated by the kind and time duration of the interaction itself. We achieved a congruent and consistent statistical significant difference for all of the three indices. Moreover, a Nearest Mean Classifier was able to recognize the three classes of interaction with an accuracy greater than 70%. Although preliminary, these encouraging results allow a discrimination of three distinct phases in a real human-animal interaction opening to the characterization of the empirically proven relationship between human and horse.

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...

Quantitative measurement of zinc secretion from pancreatic islets with high temporal resolution using droplet-based microfluidics.

Science.gov (United States)

Easley, Christopher J; Rocheleau, Jonathan V; Head, W Steven; Piston, David W

2009-11-01

We assayed glucose-stimulated insulin secretion (GSIS) from live, murine islets of Langerhans in microfluidic devices by the downstream formation of aqueous droplets. Zinc ions, which are cosecreted with insulin from beta-cells, were quantitatively measured from single islets with high temporal resolution using a fluorescent indicator, FluoZin-3. Real-time storage of secretions into droplets (volume of 0.470 +/- 0.009 nL) effectively preserves the temporal chemical information, allowing reconstruction of the secretory time record. The use of passive flow control within the device removes the need for syringe pumps, requiring only a single hand-held syringe. Under stimulatory glucose levels (11 mM), bursts of zinc as high as approximately 800 fg islet(-1) min(-1) were measured. Treatment with diazoxide effectively blocked zinc secretion, as expected. High temporal resolution reveals two major classes of oscillations in secreted zinc, with predominate periods at approximately 20-40 s and approximately 5-10 min. The more rapid oscillation periods match closely with those of intraislet calcium oscillations, while the slower oscillations are consistent with insulin pulses typically measured in bulk islet experiments or in the bloodstream. This droplet sampling technique should be widely applicable to time-resolved cellular secretion measurements, either in real-time or for postprocessing.

Timed function tests, motor function measure, and quantitative thigh muscle MRI in ambulant children with Duchenne muscular dystrophy: A cross-sectional analysis.

Science.gov (United States)

Schmidt, Simone; Hafner, Patricia; Klein, Andrea; Rubino-Nacht, Daniela; Gocheva, Vanya; Schroeder, Jonas; Naduvilekoot Devasia, Arjith; Zuesli, Stephanie; Bernert, Guenther; Laugel, Vincent; Bloetzer, Clemens; Steinlin, Maja; Capone, Andrea; Gloor, Monika; Tobler, Patrick; Haas, Tanja; Bieri, Oliver; Zumbrunn, Thomas; Fischer, Dirk; Bonati, Ulrike

2018-01-01

The development of new therapeutic agents for the treatment of Duchenne muscular dystrophy has put a focus on defining outcome measures most sensitive to capture treatment effects. This cross-sectional analysis investigates the relation between validated clinical assessments such as the 6-minute walk test, motor function measure and quantitative muscle MRI of thigh muscles in ambulant Duchenne muscular dystrophy patients, aged 6.5 to 10.8 years (mean 8.2, SD 1.1). Quantitative muscle MRI included the mean fat fraction using a 2-point Dixon technique, and transverse relaxation time (T2) measurements. All clinical assessments were highly significantly inter-correlated with p muscle MRI values significantly correlated with all clinical assessments with the extensors showing the strongest correlation. In contrast to the clinical assessments, quantitative muscle MRI values were highly significantly correlated with age. In conclusion, the motor function measure and timed function tests measure disease severity in a highly comparable fashion and all tests correlated with quantitative muscle MRI values quantifying fatty muscle degeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

Real-time quantitative PCR for retrovirus-like particle quantification in CHO cell culture.

Science.gov (United States)

de Wit, C; Fautz, C; Xu, Y

2000-09-01

Chinese hamster ovary (CHO) cells have been widely used to manufacture recombinant proteins intended for human therapeutic uses. Retrovirus-like particles, which are apparently defective and non-infectious, have been detected in all CHO cells by electron microscopy (EM). To assure viral safety of CHO cell-derived biologicals, quantification of retrovirus-like particles in production cell culture and demonstration of sufficient elimination of such retrovirus-like particles by the down-stream purification process are required for product market registration worldwide. EM, with a detection limit of 1x10(6) particles/ml, is the standard retrovirus-like particle quantification method. The whole process, which requires a large amount of sample (3-6 litres), is labour intensive, time consuming, expensive, and subject to significant assay variability. In this paper, a novel real-time quantitative PCR assay (TaqMan assay) has been developed for the quantification of retrovirus-like particles. Each retrovirus particle contains two copies of the viral genomic particle RNA (pRNA) molecule. Therefore, quantification of retrovirus particles can be achieved by quantifying the pRNA copy number, i.e. every two copies of retroviral pRNA is equivalent to one retrovirus-like particle. The TaqMan assay takes advantage of the 5'-->3' exonuclease activity of Taq DNA polymerase and utilizes the PRISM 7700 Sequence Detection System of PE Applied Biosystems (Foster City, CA, U.S.A.) for automated pRNA quantification through a dual-labelled fluorogenic probe. The TaqMan quantification technique is highly comparable to the EM analysis. In addition, it offers significant advantages over the EM analysis, such as a higher sensitivity of less than 600 particles/ml, greater accuracy and reliability, higher sample throughput, more flexibility and lower cost. Therefore, the TaqMan assay should be used as a substitute for EM analysis for retrovirus-like particle quantification in CHO cell

A quantitative method to track protein translocation between intracellular compartments in real-time in live cells using weighted local variance image analysis.

Directory of Open Access Journals (Sweden)

Guillaume Calmettes

Full Text Available The genetic expression of cloned fluorescent proteins coupled to time-lapse fluorescence microscopy has opened the door to the direct visualization of a wide range of molecular interactions in living cells. In particular, the dynamic translocation of proteins can now be explored in real time at the single-cell level. Here we propose a reliable, easy-to-implement, quantitative image processing method to assess protein translocation in living cells based on the computation of spatial variance maps of time-lapse images. The method is first illustrated and validated on simulated images of a fluorescently-labeled protein translocating from mitochondria to cytoplasm, and then applied to experimental data obtained with fluorescently-labeled hexokinase 2 in different cell types imaged by regular or confocal microscopy. The method was found to be robust with respect to cell morphology changes and mitochondrial dynamics (fusion, fission, movement during the time-lapse imaging. Its ease of implementation should facilitate its application to a broad spectrum of time-lapse imaging studies.

Development of a real-time quantitative assay for detection of Epstein-Barr virus

NARCIS (Netherlands)

Niesters, H. G.; van Esser, J.; Fries, E.; Wolthers, K. C.; Cornelissen, J.; Osterhaus, A. D.

2000-01-01

With the use of real-time PCR, we developed and evaluated a rapid, sensitive, specific, and reproducible method for the detection of Epstein-Barr virus (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and 10(7) copies of DNA per ml using

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...

DEVELOPMENT OF REAL-TIME MULTIPLEX PCR FOR THE QUANTITATIVE DETERMINATION OF TREC'S AND KREC'S IN WHOLE BLOOD AND IN DRIED BLOOD SPOTS

Directory of Open Access Journals (Sweden)

M. A. Gordukova

2015-01-01

Full Text Available Primary immunodeficiencies (PID such as severe combined immunodeficiency (SCID and X-linked agammaglobulinemia are characterized by the lack of functional Tand B-cells, respectively. Without early diagnosis and prompt treatment children with PID suffer from severe infectious diseases, leading to their death or disability. Our purpose was developing of simple, inexpensive, high throughput technique based on the quantitative determination of TREC and KREC molecules by real-time PCR, and its validation in a group of children with a verified diagnosis of SCID and X-linked agammaglobulinemia.In this study, we developed and validated multiplex real-time PCR for the TREC’s and KREC’s quantitative analysis. We have shown that linear range of Ct changes depending on the concentrations of targets with a correlation coefficient R2 not worse than 0.98 was observed at concentrations from 109 to 5 × 104 copies per ml. The lowest amount of targets reliably detected in a reaction volume was 10 TREC’s copies, 5 KREC ‘s copies and 5 copies of internal control (IL17RA. We determined the age-depended reference values of TRECs and KRECs in whole blood in 29 boys and 27 girls with normal immunological parameters. The normal cut-offs for TRECs and KRECs were defined in dry blood spots depending on the method of extraction.The proposed method showed 100% diagnostic sensitivity and specificity in the studied group. The method can be proposed as a screening tool for the diagnosis of SCID and X-linked agammaglobulinemia both in whole blood and in the dry blood spots. The further investigation is required with larger number of samples. 

Field Installation and Real-Time Data Processing of the New Integrated SeismoGeodetic System with Real-Time Acceleration and Displacement Measurements for Earthquake Characterization Based on High-Rate Seismic and GPS Data

Science.gov (United States)

Zimakov, Leonid; Jackson, Michael; Passmore, Paul; Raczka, Jared; Alvarez, Marcos; Barrientos, Sergio

2015-04-01

We will discuss and show the results obtained from an integrated SeismoGeodetic System, model SG160-09, installed in the Chilean National Network. The SG160-09 provides the user high rate GNSS and accelerometer data, full epoch-by-epoch measurement integrity and, using the Trimble Pivot™ SeismoGeodetic App, the ability to create combined GNSS and accelerometer high-rate (200Hz) displacement time series in real-time. The SG160-09 combines seismic recording with GNSS geodetic measurement in a single compact, ruggedized package. The system includes a low-power, 220-channel GNSS receiver powered by the latest Trimble-precise Maxwell™6 technology and supports tracking GPS, GLONASS and Galileo signals. The receiver incorporates on-board GNSS point positioning using Real-Time Precise Point Positioning (PPP) technology with satellite clock and orbit corrections delivered over IP networks. The seismic recording element includes an ANSS Class A, force balance triaxial accelerometer with the latest, low power, 24-bit A/D converter, which produces high-resolution seismic data. The SG160-09 processor acquires and packetizes both seismic and geodetic data and transmits it to the central station using an advanced, error-correction protocol with back fill capability providing data integrity between the field and the processing center. The SG160-09 has been installed in the seismic station close to the area of the Iquique earthquake of April 1, 2014, in northern Chile, a seismically prone area at the current time. The hardware includes the SG160-09 system, external Zephyr Geodetic-2 GNSS antenna, and high-speed Internet communication media. Both acceleration and displacement data was transmitted in real-time to the National Seismological Center in Santiago for real-time data processing using Earthworm / Early Bird software. Command/Control of the field station and real-time GNSS position correction are provided via the Pivot software suite. Data from the SG160-09 system was

Beneficial aspects of real time flow measurements for the management of acute right ventricular heart failure following continuous flow ventricular assist device implantation

Directory of Open Access Journals (Sweden)

Spiliopoulos Sotirios

2012-11-01

Full Text Available Abstract Background Optimal management of acute right heart failure following the implantation of a left ventricular assist device requires a reliable estimation of left ventricular preload and contractility. This is possible by real-time pump blood flow measurements. Clinical case We performed implantation of a continuous flow left ventricular assist device in a 66 years old female patient with an end-stage heart failure on the grounds of a dilated cardiomyopathy. Real-time pump blood flow was directly measured by an ultrasonic flow probe placed around the outflow graft. Diagnosis The progressive decline of real time flow and the loss of pulsatility were associated with an increase of central venous pressure, inotropic therapy and progressive renal failure suggesting the presence of an acute right heart failure. Diagnosis was validated by echocardiography and thermodilution measurements. Treatment Temporary mechanical circulatory support of the right ventricle was successfully performed. Real time flow measurement proved to be a useful tool for the diagnosis and ultimately for the management of right heart failure including the weaning from extracorporeal membrane oxygenation.

Real time chromametry measurement for food quality detection using mobile device

Science.gov (United States)

Witjaksono, Gunawan; Mohamad Hussin, Nur Haziqah Farah Binti; Abdelkreem Saeed Rabih, Almur; Alfa, Sagir

2017-09-01

Freshness of the food is the main factor in determining the quality and safety of the consumed food and hence consumers satisfaction. Current technologies for food quality determination depend on colour changing labels to indicate the freshness level, which is subjective to human eyes. The goal of this paper is to design and develop chromatic algorithm based on RGB colour reading and correlation with pH values for real time determination of freshness level of shrimp. The results show that the developed algorithm is able to measure, analyse and display the freshness level of food directly on the screen of a mobile app technology. The mobile app is developed on Android platform and is tested in the shrimp freshness range by stating whether it is “fresh, good or spoiled”.

Linux real-time framework for fusion devices

Energy Technology Data Exchange (ETDEWEB)

Neto, Andre [Associacao Euratom-IST, Instituto de Plasmas e Fusao Nuclear, Av. Rovisco Pais, 1049-001 Lisboa (Portugal)], E-mail: andre.neto@cfn.ist.utl.pt; Sartori, Filippo; Piccolo, Fabio [Euratom-UKAEA, Culham Science Centre, Abingdon, Oxon OX14 3DB (United Kingdom); Barbalace, Antonio [Euratom-ENEA Association, Consorzio RFX, 35127 Padova (Italy); Vitelli, Riccardo [Dipartimento di Informatica, Sistemi e Produzione, Universita di Roma, Tor Vergata, Via del Politecnico 1-00133, Roma (Italy); Fernandes, Horacio [Associacao Euratom-IST, Instituto de Plasmas e Fusao Nuclear, Av. Rovisco Pais, 1049-001 Lisboa (Portugal)

2009-06-15

A new framework for the development and execution of real-time codes is currently being developed and commissioned at JET. The foundations of the system are Linux, the Real Time Application Interface (RTAI) and a wise exploitation of the new i386 multi-core processors technology. The driving motivation was the need to find a real-time operating system for the i386 platform able to satisfy JET Vertical Stabilisation Enhancement project requirements: 50 {mu}s cycle time. Even if the initial choice was the VxWorks operating system, it was decided to explore an open source alternative, mostly because of the costs involved in the commercial product. The work started with the definition of a precise set of requirements and milestones to achieve: Linux distribution and kernel versions to be used for the real-time operating system; complete characterization of the Linux/RTAI real-time capabilities; exploitation of the multi-core technology; implementation of all the required and missing features; commissioning of the system. Latency and jitter measurements were compared for Linux and RTAI in both user and kernel-space. The best results were attained using the RTAI kernel solution where the time to reschedule a real-time task after an external interrupt is of 2.35 {+-} 0.35 {mu}s. In order to run the real-time codes in the kernel-space, a solution to provide user-space functionalities to the kernel modules had to be designed. This novel work provided the most common functions from the standard C library and transparent interaction with files and sockets to the kernel real-time modules. Kernel C++ support was also tested, further developed and integrated in the framework. The work has produced very convincing results so far: complete isolation of the processors assigned to real-time from the Linux non real-time activities, high level of stability over several days of benchmarking operations and values well below 3 {mu}s for task rescheduling after external interrupt. From

Linux real-time framework for fusion devices

International Nuclear Information System (INIS)

Neto, Andre; Sartori, Filippo; Piccolo, Fabio; Barbalace, Antonio; Vitelli, Riccardo; Fernandes, Horacio

2009-01-01

A new framework for the development and execution of real-time codes is currently being developed and commissioned at JET. The foundations of the system are Linux, the Real Time Application Interface (RTAI) and a wise exploitation of the new i386 multi-core processors technology. The driving motivation was the need to find a real-time operating system for the i386 platform able to satisfy JET Vertical Stabilisation Enhancement project requirements: 50 μs cycle time. Even if the initial choice was the VxWorks operating system, it was decided to explore an open source alternative, mostly because of the costs involved in the commercial product. The work started with the definition of a precise set of requirements and milestones to achieve: Linux distribution and kernel versions to be used for the real-time operating system; complete characterization of the Linux/RTAI real-time capabilities; exploitation of the multi-core technology; implementation of all the required and missing features; commissioning of the system. Latency and jitter measurements were compared for Linux and RTAI in both user and kernel-space. The best results were attained using the RTAI kernel solution where the time to reschedule a real-time task after an external interrupt is of 2.35 ± 0.35 μs. In order to run the real-time codes in the kernel-space, a solution to provide user-space functionalities to the kernel modules had to be designed. This novel work provided the most common functions from the standard C library and transparent interaction with files and sockets to the kernel real-time modules. Kernel C++ support was also tested, further developed and integrated in the framework. The work has produced very convincing results so far: complete isolation of the processors assigned to real-time from the Linux non real-time activities, high level of stability over several days of benchmarking operations and values well below 3 μs for task rescheduling after external interrupt. From being the

Opportunities for measuring DNA synthesis time by quantitative autoradiography

International Nuclear Information System (INIS)

Vasileva, D.

1980-01-01

DNA sysntesis time (Tsub(s)) in cells of the canine erythropoiesis and myelopoiesis pools was determined by quantitative autoradiography according to Doermer. In contrast to mitosis labelling for Tsub(s) estimation as so far applied, this technique uses well-differentiated cells. After blocking endogeneous DNA synthesis with 5-fluorodeoxyuridine, its further course becomes dependent on exogeneous supply of thymidine, in the form of 14 C-thymidine. From incroporation of the latter into the individual cell within a definite time span (3-7 min) and taking into account its total amount, Tsub(s) may be calculated. The data thus obtained were found to agree with Tsub(s) values as estimated from the labelled mitosis curve

A two-domain real-time algorithm for optimal data reduction: a case study on accelerator magnet measurements

International Nuclear Information System (INIS)

Arpaia, Pasquale; Buzio, Marco; Inglese, Vitaliano

2010-01-01

A real-time algorithm of data reduction, based on the combination of two lossy techniques specifically optimized for high-rate magnetic measurements in two domains (e.g. time and space), is proposed. The first technique exploits an adaptive sampling rule based on the power estimation of the flux increments in order to optimize the information to be gathered for magnetic field analysis in real time. The tracking condition is defined by the target noise level in the Nyquist band required by the post-processing procedure of magnetic analysis. The second technique uses a data reduction algorithm in order to improve the compression ratio while preserving the consistency of the measured signal. The allowed loss is set equal to the random noise level in the signal in order to force the loss and the noise to cancel rather than to add, by improving the signal-to-noise ratio. Numerical analysis and experimental results of on-field performance characterization and validation for two case studies of magnetic measurement systems for testing magnets of the Large Hadron Collider at the European Organization for Nuclear Research (CERN) are reported

RTX Correction Accuracy and Real-Time Data Processing of the New Integrated SeismoGeodetic System with Real-Time Acceleration and Displacement Measurements for Earthquake Characterization Based on High-Rate Seismic and GPS Data

Science.gov (United States)

Zimakov, L. G.; Raczka, J.; Barrientos, S. E.

2016-12-01

We will discuss and show the results obtained from an integrated SeismoGeodetic System, model SG160-09, installed in the Chile (Chilean National Network), Italy (University of Naples Network), and California. The SG160-09 provides the user high rate GNSS and accelerometer data, full epoch-by-epoch measurement integrity and the ability to create combined GNSS and accelerometer high-rate (200Hz) displacement time series in real-time. The SG160-09 combines seismic recording with GNSS geodetic measurement in a single compact, ruggedized case. The system includes a low-power, 220-channel GNSS receiver powered by the latest Trimble-precise Maxwell™6 technology and supports tracking GPS, GLONASS and Galileo signals. The receiver incorporates on-board GNSS point positioning using Real-Time Precise Point Positioning (PPP) technology with satellite clock and orbit corrections delivered over IP networks. The seismic recording includes an ANSS Class A, force balance accelerometer with the latest, low power, 24-bit A/D converter, producing high-resolution seismic data. The SG160-09 processor acquires and packetizes both seismic and geodetic data and transmits it to the central station using an advanced, error-correction protocol providing data integrity between the field and the processing center. The SG160-09 has been installed in three seismic stations in different geographic locations with different Trimble global reference stations coverage The hardware includes the SG160-09 system, external Zephyr Geodetic-2 GNSS antenna, both radio and high-speed Internet communication media. Both acceleration and displacement data was transmitted in real-time to the centralized Data Acquisition Centers for real-time data processing. Command/Control of the field station and real-time GNSS position correction are provided via the Pivot platform. Data from the SG160-09 system was used for seismic event characterization along with data from traditional seismic and geodetic stations

A quantitative method for measuring the quality of history matches

Energy Technology Data Exchange (ETDEWEB)

Shaw, T.S. [Kerr-McGee Corp., Oklahoma City, OK (United States); Knapp, R.M. [Univ. of Oklahoma, Norman, OK (United States)

1997-08-01

History matching can be an efficient tool for reservoir characterization. A {open_quotes}good{close_quotes} history matching job can generate reliable reservoir parameters. However, reservoir engineers are often frustrated when they try to select a {open_quotes}better{close_quotes} match from a series of history matching runs. Without a quantitative measurement, it is always difficult to tell the difference between a {open_quotes}good{close_quotes} and a {open_quotes}better{close_quotes} matches. For this reason, we need a quantitative method for testing the quality of matches. This paper presents a method for such a purpose. The method uses three statistical indices to (1) test shape conformity, (2) examine bias errors, and (3) measure magnitude of deviation. The shape conformity test insures that the shape of a simulated curve matches that of a historical curve. Examining bias errors assures that model reservoir parameters have been calibrated to that of a real reservoir. Measuring the magnitude of deviation assures that the difference between the model and the real reservoir parameters is minimized. The method was first tested on a hypothetical model and then applied to published field studies. The results showed that the method can efficiently measure the quality of matches. It also showed that the method can serve as a diagnostic tool for calibrating reservoir parameters during history matching.

Real-time statistical quality control and ARM

International Nuclear Information System (INIS)

Blough, D.K.

1992-05-01

An important component of the Atmospheric Radiation Measurement (ARM) Program is real-time quality control of data obtained from meteorological instruments. It is the goal of the ARM program to enhance the predictive capabilities of global circulation models by incorporating in them more detailed information on the radiative characteristics of the earth's atmosphere. To this end, a number of Cloud and Radiation Testbeds (CART's) will be built at various locations worldwide. Each CART will consist of an array of instruments designed to collect radiative data. The large amount of data obtained from these instruments necessitates real-time processing in order to flag outliers and possible instrument malfunction. The Bayesian dynamic linear model (DLM) proves to be an effective way of monitoring the time series data which each instrument generates. It provides a flexible yet powerful approach to detecting in real-time sudden shifts in a non-stationary multivariate time series. An application of these techniques to data arising from a remote sensing instrument to be used in the CART is provided. Using real data from a wind profiler, the ability of the DLM to detect outliers is studied. 5 refs

Mid-IR spectrometer for mobile, real-time urban NO2 measurements

Science.gov (United States)

Morten Hundt, P.; Müller, Michael; Mangold, Markus; Tuzson, Béla; Scheidegger, Philipp; Looser, Herbert; Hüglin, Christoph; Emmenegger, Lukas

2018-05-01

Detailed knowledge about the urban NO2 concentration field is a key element for obtaining accurate pollution maps and individual exposure estimates. These are required for improving the understanding of the impact of ambient NO2 on human health and for related air quality measures. However, city-scale NO2 concentration maps with high spatio-temporal resolution are still lacking, mainly due to the difficulty of accurate measurement of NO2 at the required sub-ppb level precision. We contribute to close this gap through the development of a compact instrument based on mid-infrared laser absorption spectroscopy. Leveraging recent advances in infrared laser and detection technology and a novel circular absorption cell, we demonstrate the feasibility and robustness of this technique for demanding mobile applications. A fully autonomous quantum cascade laser absorption spectrometer (QCLAS) has been successfully deployed on a tram, performing long-term and real-time concentration measurements of NO2 in the city of Zurich (Switzerland). For ambient NO2 concentrations, the instrument demonstrated a precision of 0.23 ppb at one second time resolution and of 0.03 ppb after 200 s averaging. Whilst the combined uncertainty estimated for the retrieved spectroscopic values was less than 5 %, laboratory intercomparison measurements with standard CLD instruments revealed a systematic NO2 wall loss of about 10 % within the laser spectrometer. For the field campaign, the QCLAS has been referenced to a CLD using urban atmospheric air, despite the potential cross sensitivity of CLD to other nitrogen containing compounds. However, this approach allowed a direct comparison and continuous validation of the spectroscopic data to measurements at regulatory air quality monitoring (AQM) stations along the tram-line. The analysis of the recorded high-resolution time series allowed us to gain more detailed insights into the spatio-temporal concentration distribution of NO2 in an urban