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Sample records for real-time quantitative measurement

  1. Quantitative density measurements from a real-time neutron radiography system

    International Nuclear Information System (INIS)

    McRae, D.D.; Jenkins, R.W. Jr.; Brenizer, J.S.; Tobin, K.W.; Hosticka, B.; Sulcoski, M.F.

    1986-01-01

    An advanced video system has been assembled from commercially available equipment to support the real-time neutron radiography facility established jointly by the University of Virginia Department of Nuclear Engineering and Engineering Physics, and the Philip Morris Research Center. A schematic diagram of the equipment used for real-time neutron radiography is presented. To obtain quantitative density measurements with this system, several modifications of both hardware and image processing software were required. After implementation of these changes, the system was capable of determining material densities by measuring the degree of neutron attenuation

  2. Quantitative (real-time) PCR

    International Nuclear Information System (INIS)

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

    2005-01-01

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

  3. 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

  4. Quantitative analysis of real-time radiographic systems

    International Nuclear Information System (INIS)

    Barker, M.D.; Condon, P.E.; Barry, R.C.; Betz, R.A.; Klynn, L.M.

    1988-01-01

    A method was developed which yields quantitative information on the spatial resolution, contrast sensitivity, image noise, and focal spot size from real time radiographic images. The method uses simple image quality indicators and computer programs which make it possible to readily obtain quantitative performance measurements of single or multiple radiographic systems. It was used for x-ray and optical images to determine which component of the system was not operating up to standard. Focal spot size was monitored by imaging a bar pattern. This paper constitutes the second progress report on the development of the camera and radiation image quality indicators

  5. 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

  6. Diagnosis of aerobic vaginitis by quantitative real-time PCR

    OpenAIRE

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

    2016-01-01

    Abstract: Purpose To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. Methods Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. Resu...

  7. 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

  8. 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.

  9. 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.

  10. Human fecal source identification with real-time quantitative PCR

    Science.gov (United States)

    Waterborne diseases represent a significant public health risk worldwide, and can originate from contact with water contaminated with human fecal material. We describe a real-time quantitative PCR (qPCR) method that targets a Bacteroides dori human-associated genetic marker for...

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Real time measurement of air radioactivity

    International Nuclear Information System (INIS)

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

    1998-01-01

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

  18. Superhilac real-time velocity measurements

    International Nuclear Information System (INIS)

    Feinberg, B.; Meaney, D.; Thatcher, R.; Timossi, C.

    1987-03-01

    Phase probes have been placed in several external beam lines at the LBL heavy ion linear accelerator (SuperHILAC) to provide non-destructive velocity measurements independent of the ion being accelerated. The existing system has been improved to provide the following features: a display refresh rate better than twice per second, a sensitive pseudo-correlation technique to pick out the signal from the noise, simultaneous measurements of up to four ion velocities when more than one beam is being accelerated, and a touch-screen operator interface. These improvements allow the system to be used as a routine tuning aid and beam velocity monitor

  19. 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

  20. 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

  1. 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)

  2. 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)

  3. Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR.

    NARCIS (Netherlands)

    Pierik, A.; Moamfa, M; van Zelst, M.; Clout, D.; Stapert, H.; Dijksman, Johan Frederik; Broer, D.; Wimberger-Friedl, R.

    2012-01-01

    Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that

  4. Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR

    NARCIS (Netherlands)

    Pierik, Anke; Boamfa, M.; Zelst, van M.; Clout, D.; Stapert, H.R.; Dijksman, J.F.; Broer, D.J.; Wimberger-Friedl, R.

    2012-01-01

    Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that

  5. 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.)

  6. A Quantitative Approach to the Formal Verification of Real-Time Systems.

    Science.gov (United States)

    1996-09-01

    Computer Science A Quantitative Approach to the Formal Verification of Real - Time Systems Sergio Vale Aguiar Campos September 1996 CMU-CS-96-199...ptisiic raieaiSI v Diambimos Lboiamtad _^ A Quantitative Approach to the Formal Verification of Real - Time Systems Sergio Vale Aguiar Campos...implied, of NSF, the Semiconduc- tor Research Corporation, ARPA or the U.S. government. Keywords: real - time systems , formal verification, symbolic

  7. 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)

  8. 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.

  9. 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

  10. 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

  11. 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

  12. Quantitative real-time monitoring of dryer effluent using fiber optic near-infrared spectroscopy.

    Science.gov (United States)

    Harris, S C; Walker, D S

    2000-09-01

    This paper describes a method for real-time quantitation of the solvents evaporating from a dryer. The vapor stream in the vacuum line of a dryer was monitored in real time using a fiber optic-coupled acousto-optic tunable filter near-infrared (AOTF-NIR) spectrometer. A balance was placed in the dryer, and mass readings were recorded for every scan of the AOTF-NIR. A partial least-squares (PLS) calibration was subsequently built based on change in mass over change in time for solvents typically used in a chemical manufacturing plant. Controlling software for the AOTF-NIR was developed. The software collects spectra, builds the PLS calibration model, and continuously fits subsequently collected spectra to the calibration, allowing the operator to follow the mass loss of solvent from the dryer. The results indicate that solvent loss can be monitored and quantitated in real time using NIR for the optimization of drying times. These time-based mass loss values have also been used to calculate "dynamic" vapor density values for the solvents. The values calculated are in agreement with values determined from the ideal gas law and could prove valuable as tools to measure temperature or pressure indirectly.

  13. Real-time quantitative phase reconstruction in off-axis digital holography using multiplexing.

    Science.gov (United States)

    Girshovitz, Pinhas; Shaked, Natan T

    2014-04-15

    We present a new approach for obtaining significant speedup in the digital processing of extracting unwrapped phase profiles from off-axis digital holograms. The new technique digitally multiplexes two orthogonal off-axis holograms, where the digital reconstruction, including spatial filtering and two-dimensional phase unwrapping on a decreased number of pixels, can be performed on both holograms together, without redundant operations. Using this technique, we were able to reconstruct, for the first time to our knowledge, unwrapped phase profiles from off-axis holograms with 1 megapixel in more than 30 frames per second using a standard single-core personal computer on a MATLAB platform, without using graphic-processing-unit programming or parallel computing. This new technique is important for real-time quantitative visualization and measurements of highly dynamic samples and is applicable for a wide range of applications, including rapid biological cell imaging and real-time nondestructive testing. After comparing the speedups obtained by the new technique for holograms of various sizes, we present experimental results of real-time quantitative phase visualization of cells flowing rapidly through a microchannel.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. Quantification bias caused by plasmid DNA conformation in quantitative real-time PCR assay.

    Science.gov (United States)

    Lin, Chih-Hui; Chen, Yu-Chieh; Pan, Tzu-Ming

    2011-01-01

    Quantitative real-time PCR (qPCR) is the gold standard for the quantification of specific nucleic acid sequences. However, a serious concern has been revealed in a recent report: supercoiled plasmid standards cause significant over-estimation in qPCR quantification. In this study, we investigated the effect of plasmid DNA conformation on the quantification of DNA and the efficiency of qPCR. Our results suggest that plasmid DNA conformation has significant impact on the accuracy of absolute quantification by qPCR. DNA standard curves shifted significantly among plasmid standards with different DNA conformations. Moreover, the choice of DNA measurement method and plasmid DNA conformation may also contribute to the measurement error of DNA standard curves. Due to the multiple effects of plasmid DNA conformation on the accuracy of qPCR, efforts should be made to assure the highest consistency of plasmid standards for qPCR. Thus, we suggest that the conformation, preparation, quantification, purification, handling, and storage of standard plasmid DNA should be described and defined in the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) to assure the reproducibility and accuracy of qPCR absolute quantification.

  19. Complementary techniques: validation of gene expression data by quantitative real time PCR.

    Science.gov (United States)

    Provenzano, Maurizio; Mocellin, Simone

    2007-01-01

    Microarray technology can be considered the most powerful tool for screening gene expression profiles of biological samples. After data mining, results need to be validated with highly reliable biotechniques allowing for precise quantitation of transcriptional abundance of identified genes. Quantitative real time PCR (qrt-PCR) technology has recently reached a level of sensitivity, accuracy and practical ease that support its use as a routine bioinstrumentation for gene level measurement. Currently, qrt-PCR is considered by most experts the most appropriate method to confirm or confute microarray-generated data. The knowledge of the biochemical principles underlying qrt-PCR as well as some related technical issues must be beard in mind when using this biotechnology.

  20. 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.

  1. 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.

  2. Real-time quantitative PCR of microdissected paraffin-embedded breast carcinoma

    DEFF Research Database (Denmark)

    Gjerdrum, Lise Mette; Sorensen, Boe Sandahl; Kjeldsen, Eigil

    2004-01-01

    We studied the feasibility of using real-time quantitative PCR to determine HER-2 DNA amplification and mRNA expression in microdissected formalin-fixed, paraffin-embedded breast tumors and compared this with standard immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) methods...... tumors as being amplified. Interestingly, all these scored 2+ with the HercepTest, but were negative using FISH. We believe that real-time quantitative PCR analysis of HER-2 DNA amplification following microdissection represents a useful supplementary or perhaps even an alternative technique...

  3. 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.

  4. 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

  5. Real-time quantitative PCR of microdissected paraffin-embedded breast carcinoma

    DEFF Research Database (Denmark)

    Gjerdrum, Lise Mette; Sorensen, Boe Sandahl; Kjeldsen, Eigil

    2004-01-01

    We studied the feasibility of using real-time quantitative PCR to determine HER-2 DNA amplification and mRNA expression in microdissected formalin-fixed, paraffin-embedded breast tumors and compared this with standard immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) methods...

  6. Quantitative real-time RT-PCR and chromogenic in situ hybridization

    DEFF Research Database (Denmark)

    Rosa, Fabíola E; Silveira, Sara M; Silveira, Cássia G T

    2009-01-01

    . METHODS: To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. RESULTS: The concordance...

  7. 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

  8. Real-time quantitative PCR of Staphylococcus aureus and application in restaurant meals.

    Science.gov (United States)

    Berrada, H; Soriano, J M; Mañes, J; Picó, Y

    2006-01-01

    Staphylococcus aureus is considered the second most common pathogen to cause outbreaks of food poisoning, exceeded only by Campylobacter. Consumption of foods containing this microorganism is often identified as the cause of illness. In this study, a rapid, reliable, and sensitive real-time quantitative PCR was developed and compared with conventional culture methods. Real-time quantitative PCR was carried out by purifying DNA extracts of S. aureus with a Staphylococcus sample preparation kit and quantifying it in the LightCycler system with hybridization probes. The assay was linear from a range of 10 to 10(6) S. aureus cells (r2 > 0.997). The PCR reaction presented an efficiency of >85%. Accuracy of the PCR-based assay, expressed as percent bias, was around 13%, and the precision, expressed as a percentage of the coefficient of variation, was 7 to 10%. Intraday and interday variability were studied at 10(2) CFU/g and was 12 and 14%, respectively. The proposed method was applied to the analysis of 77 samples of restaurant meals in Valencia (Spain). In 11.6% of samples S. aureus was detected by real-time quantitative PCR, as well as by the conventional microbiological method. An excellent correspondence between real-time quantitative PCR and microbiological numbers (CFU/g) was observed with deviations of < 28%.

  9. 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.

  10. 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....

  11. 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....

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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...

  2. 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...

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Real time quantitative phase microscopy based on single-shot transport of intensity equation (ssTIE) method

    Science.gov (United States)

    Yu, Wei; Tian, Xiaolin; He, Xiaoliang; Song, Xiaojun; Xue, Liang; Liu, Cheng; Wang, Shouyu

    2016-08-01

    Microscopy based on transport of intensity equation provides quantitative phase distributions which opens another perspective for cellular observations. However, it requires multi-focal image capturing while mechanical and electrical scanning limits its real time capacity in sample detections. Here, in order to break through this restriction, real time quantitative phase microscopy based on single-shot transport of the intensity equation method is proposed. A programmed phase mask is designed to realize simultaneous multi-focal image recording without any scanning; thus, phase distributions can be quantitatively retrieved in real time. It is believed the proposed method can be potentially applied in various biological and medical applications, especially for live cell imaging.

  9. DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I

    DEFF Research Database (Denmark)

    Marcussen, Lærke Bay; Jepsen, Morten Leth; Kristoffersen, Emil Laust

    2013-01-01

    Sensors capable of quantitative real-time measurements may present the easiest and most accurate way to study enzyme activities. Here we present a novel DNA-based sensor for specific and quantitative real-time measurement of the enzymatic activity of the essential human enzyme, topoisomerase I....... The basic design of the sensor relies on two DNA strands that hybridize to form a hairpin structure with a fluorophore-quencher pair. The quencher moiety is released from the sensor upon reaction with human topoisomerase I thus enabling real-time optical measurement of enzymatic activity. The sensor....... The cytotoxic effect of camptothecins correlates directly with the intracellular topoisomerase I activity. We therefore envision that the presented sensor may find use for the prediction of cellular drug response. Moreover, inhibition of topoisomerase I by camptothecin is readily detectable using the presented...

  10. A RECURSIVE ALGORITHM SUITABLE FOR REAL-TIME MEASUREMENT

    Directory of Open Access Journals (Sweden)

    Giovanni Bucci

    1995-12-01

    Full Text Available This paper deals with a recursive algorithm suitable for realtime measurement applications, based on an indirect technique, useful in those applications where the required quantities cannot be measured in a straightforward way. To cope with time constraints a parallel formulation of it, suitable to be implemented on multiprocessor systems, is presented. The adopted concurrent implementation is based on factorization techniques. Some experimental results related to the application of the system for carrying out measurements on synchronous motors are included.

  11. Real time wave measurements and wave hindcasting in deep waters

    Digital Repository Service at National Institute of Oceanography (India)

    Anand, N.M.; Mandal, S.; SanilKumar, V.; Nayak, B.U.

    Deep water waves off Karwar (lat. 14~'45.1'N, long. 73~'34.8'E) at 75 m water depth pertaining to peak monsoon period have been measured using a Datawell waverider buoy. Measured wave data show that the significant wave height (Hs) predominantly...

  12. 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.

  13. Real-time precision measuring device of tree diameter growth

    Science.gov (United States)

    Guo, Mingming; Chen, Aijun; Li, Dongsheng; Liu, Nan; Yao, Jingyuan

    2016-01-01

    DBH(diameter at breast height) is an important factor to reflect of the quality of plant growth, also an important parameter indispensable in forest resources inventory and forest carbon sink, the accurate measurement of DBH or not is directly related to the research of forest resources inventory and forest carbon sink. In this paper, the principle and the mathematical model of DBH measurement device were introduced, the fixture measuring device and the hardware circuit for this tree diameter were designed, the measurement software programs were compiled, and the precision measuring device of tree diameter growth was developed. Some experiments with Australia fir were conducted. Based on experiment data, the correlations among the DBH variation of Australian fir, the environment temperature, air humility and PAR(photosynthetically active radiation) were obtained. The effects of environmental parameters (environment temperature, air humility and PAR) on tree diameter were analyzed. Experimental results show that there is a positive correlation between DBH variation of Australian fir and environment temperature, a negative correlation between DBH variation of Australian fir and air humility , so is PAR.

  14. Real-Time Measurements for Adaptive and Cognitive Radio Systems

    Directory of Open Access Journals (Sweden)

    Hüseyin Arslan

    2009-01-01

    Full Text Available Adaptive and cognitive radios (CR have been becoming popular for optimizing mobile radio system transmission and reception. One of the most important elements of the adaptive radio and CR concepts is the ability to measure, sense, learn about, and be aware of parameters related to the radio channel characteristics, availability of spectrum and power, interference and noise temperature, operational environment of radio, user requirements and applications, available networks and infrastructures, local policies, other operating restrictions, and so on. This paper discusses some of the important measurement parameters for enabling adaptive radio and CR systems along with their relationships and impacts on the performance including relevant challenges.

  15. Application of real time spectrum measurement to radiation monitors

    International Nuclear Information System (INIS)

    Matsuno, K.; Watanabe, M.; Sakamaki, T.

    1996-01-01

    A multichannel analyzer (MCA) and two realtime spectrum monitoring methods have been developed for use in radiation monitors. The new MCA was designed to be installed at a local site as a component of a radiation monitor. The MCA repeats spectrum measurement at short intervals (Δt) and, after each measurement, transmits a spectrum datum to the operation console. The authors applied two methods to process Δt spectrum counts for each channel for longer time interval. One method of processing counts is the 'running average (RA) method'. The other method is the 'exponential smoothing (ES) method', which simulates RC rate meters by subtracting a fraction corresponding to the accumulated counts. Relative standard deviations for each channel can be made the same by selecting an appropriate value. The response with the 'ES' method is initially faster than that with the 'RA' method, but the 'RA' method allows a full response to be reached at a predictable time. (author)

  16. Microreactortechnology: Real-Time Flow Measurements in Organic Synthesis

    Directory of Open Access Journals (Sweden)

    Pieter J. Nieuwland

    2012-03-01

    Full Text Available With the commercial availability of integrated microreactor systems, the numbers of chemical processes that are performed nowadays in a continuous flow is growing rapidly. The control over mixing efficiency and homogeneous heating in these reactors allows industrial scale production that was often hampered by the use of large amounts of hazardous chemicals. Accurate actuation and in line measurements of the flows, to have a better control over the chemical reaction, is of added value for increasing reproducibility and a safe production.

  17. 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

  18. Radiation exposure in nuclear medicine: real-time measurement

    International Nuclear Information System (INIS)

    Sylvain, Iara; Bok, Bernard; X. Bichat University, Paris

    2002-01-01

    French regulations have introduced the use of electronic dosimeters for personnel monitoring of workers. In order to evaluate the exposure from diagnostic procedures to nuclear medicine staff, individual whole-body doses were measured daily with electronic (digital) personal dosimeters during 20 consecutive weeks and correlated with the work load of each day. Personal doses remained always below 20 mu Sv/d under normal working conditions. Radiation exposure levels were highest to tech staff, nurses and stretcher-bearers. The extrapolated annual cumulative doses for all staff remained less than 10% of the maximum legal limit for exposed workers (2 mSv/yr). Electronic dosimeters are not technically justified for routine survey of staff. The high sensitivity and immediate reading of electronic semiconductor dosimeters may become very useful for exposure control under risky working conditions. It may become an important help for optimising radiation protection. (author)

  19. 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...

  20. 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...

  1. 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.

  2. 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.

  3. 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.

  4. Radiation exposure in nuclear medicine: real-time measurement

    Directory of Open Access Journals (Sweden)

    Iara Sylvain

    2002-09-01

    Full Text Available French regulations have introduced the use of electronic dosimeters for personal monitoring of workers. In order to evaluate the exposure from diagnostic procedures to nuclear medicine staff, individual whole-body doses were measured daily with electronic (digital personal dosimeters during 20 consecutive weeks and correlated with the work load of each day. Personal doses remained always below 20 µSv/d under normal working conditions. Radiation exposure levels were highest to tech staff, nurses and stretcher-bearers. The extrapolated annual cumulative doses for all staff remained less than 10 % of the maximum legal limit for exposed workers (2 mSv/yr. Electronic dosimeters are not technically justified for routine survey of staff. The high sensitivity and immediate reading of electronic semiconductor dosimeters may become very useful for exposure control under risky working conditions. It may become an important help for optimising radiation protection.A legislação francesa introduziu o uso de dosímetros eletrônicos para monitoração da exposição do trabalhador. Afim de avaliar a exposição do trabalhador proveniente de exames diagnósticos em medicina nuclear, doses individuais do corpo inteiro foram medidas diariamente com dosímetros eletrônicos (digitais durante 20 semanas consecutivas e correlatas com as atividades de trabalho de cada dia. As doses foram sempre inferiores à 20 µSv por dia em condições normais de trabalho. Os níveis de exposição de radiação mais elevados foram para os enfermeiros, manipuladores e maqueiros. A extrapolação da dose anual para todos os trabalhadores foi menos que 10 % do limite máximo legal para os trabalhadores expostos (2 mSv/ano. Dosímetros eletrônicos não são tecnicamente justificados para a o controle de rotina da exposição dos trabalhadores, mas a alta sensibilidade e a leitura imediata desses dosímetros podem vir a serem muito úteis para o controle da exposição em condi

  5. Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

    Science.gov (United States)

    Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

    2015-12-14

    Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

  6. Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform

    Science.gov (United States)

    Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

    2015-11-01

    Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (r

  7. 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

  8. 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

  9. 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.

  10. 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

  11. Evaluation of Housekeeping Genes for Quantitative Real-Time PCR Analysis of Bradysia odoriphaga (Diptera: Sciaridae

    Directory of Open Access Journals (Sweden)

    Caihua Shi

    2016-07-01

    Full Text Available The soil insect Bradysia odoriphaga (Diptera: Sciaridae causes substantial damage to Chinese chive. Suitable reference genes in B. odoriphaga (Bradysia odoriphaga have yet to be identified for normalizing target gene expression among samples by quantitative real-time PCR (qRT-PCR. This study was focused on identifying the expression stability of 12 candidate housekeeping genes in B. odoriphaga under various experiment conditions. The final stability ranking of 12 housekeeping genes was obtained with RefFinder, and the most suitable number of reference genes was analyzed by GeNorm. The results revealed that the most appropriate sets of internal controls were RPS15, RPL18, and RPS18 across developmental phases; RPS15, RPL28, and GAPDH across temperatures; RPS15 and RPL18 across pesticide treatments; RSP5, RPS18, and SDHA across photoperiods; ACTb, RPS18, and RPS15 across diets; RPS13 and RPL28 across populations; and RPS15, ACTb, and RPS18 across all samples. The use of the most suitable reference genes versus an arbitrarily selected reference gene resulted in significant differences in the analysis of a target gene expression. HSP23 in B. odoriphaga was found to be up-regulated under low temperatures. These results will contribute to the standardization of qRT-PCR and will also be valuable for further research on gene function in B. odoriphaga.

  12. 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.

  13. 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.

  14. 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)

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. [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.

  2. 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

  3. 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

  4. 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.

  5. 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...

  6. 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.

  7. Ultrasensitive quantitation of human papillomavirus type 16 E6 oncogene sequences by nested real time PCR

    Directory of Open Access Journals (Sweden)

    López-Revilla Rubén

    2010-05-01

    Full Text Available Abstract Background We have developed an ultrasensitive method based on conventional PCR preamplification followed by nested amplification through real time PCR (qPCR in the presence of the DNA intercalating agent EvaGreen. Results Amplification mixtures calibrated with a known number of pHV101 copies carrying a 645 base pair (bp-long insert of the human papillomavirus type 16 (HPV16 E6 oncogene were used to generate the E6-1 amplicon of 645 bp by conventional PCR and then the E6-2 amplicon of 237 bp by nested qPCR. Direct and nested qPCR mixtures for E6-2 amplification corresponding to 2.5 × 102-2.5 × 106 initial pHV101 copies had threshold cycle (Ct values in the ranges of 18.7-29.0 and 10.0-25.0, respectively. The Ct of qPCR mixtures prepared with 1/50 volumes of preamplified mixtures containing 50 ng of DNA of the SiHa cell line (derived from an invasive cervical cancer with one HPV16 genome per cell was 19.9. Thermal fluorescence extinction profiles of E6-2 amplicons generated from pHV101 and SiHa DNA were identical, with a peak at 85.5°C. Conclusions Our method based on conventional preamplification for 15 cycles increased 10,750 times the sensitivity of nested qPCR for the quantitation of the E6 viral oncogene and confirmed that the SiHa cell line contains one E6-HPV16 copy per cell.

  8. Exploring valid reference genes for quantitative real - time rt - pce studies of hydrogenperoxide signaling in arabidopsis

    International Nuclear Information System (INIS)

    Zhou, H.; Han, B.; Xie, Y.; Zhang, J.; Shen, W.

    2015-01-01

    Hydrogen peroxide (H/sub 2/O/sub 2/ ) acts as a signaling molecule modulating the expression of various genes in plants. However, the reference gene(s) used for gene expression analysis of H/sub 2/O/sub 2/ signaling is still arbitrary. A reliable result obtained by quantitative real-time RT-PCR (RT-qPCR) highly depends on accurate transcript normalization using stably expressed reference genes, whereas the inaccurate normalization could easily lead to the false conclusions. In this report, by using geNorm and NormFinder algorithms, 12 candidate reference genes were evaluated and compared in root and shoot tissues of Arabidopsis upon different doses of H/sub 2/O/sub 2/. The results revealed that, in our experimental conditions, three novel reference genes (TIP41-like, UKN, and UBC21) were identified and validated as suitable reference genes for RT-qPCR normalization in both root and shoot tissues under oxidative stress. This conclusion was further confirmed by publicly available microarray data of methyl viologen and drought stress. In comparison with a single reference gene (EF-1a), the expression pattern of ZAT12 modulated by H/sub 2/O/sub 2/, when using TIP41-like, UKN, and UBC21 as multiple reference gene(s), was similar with the previous reports by using northern blotting. Thus, we proposed that these three reference genes might be good candidates for other researchers to include in their reference gene validation in gene expression studies under H/sub 2/O/sub 2/ related oxidative stress. (author)

  9. 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.

  10. 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.

  11. 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.)

  12. 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.

  13. 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.

  14. 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

  15. 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...

  16. 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...

  17. 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....

  18. 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...

  19. 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....

  20. 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...

  1. 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...

  2. 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

  3. On-Orbit Quantitative Real-Time Gene Expression Analysis Using the Wetlab-2 System

    Science.gov (United States)

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

    2015-01-01

    NASA Ames Research Center's WetLab-2 Project enables on-orbit quantitative Reverse Transcriptase PCR (qRT-PCR) analysis without the need for sample return. The WetLab-2 system is capable of processing sample types ranging from microbial cultures to animal tissues dissected on-orbit. The project developed a RNA preparation module that can lyse cells and extract RNA of sufficient quality and quantity for use as templates in qRT-PCR reactions. Our protocol has the advantage of using non-toxic chemicals and does not require alcohols or other organics. The resulting RNA is dispensed into reaction tubes that contain all lyophilized reagents needed to perform qRT-PCR reactions. System operations require simple and limited crew actions including syringe pushes, valve turns and pipette dispenses. The project selected the Cepheid SmartCycler (TradeMark), a Commercial-Off-The-Shelf (COTS) qRT-PCR unit, because of its advantages including rugged modular design, low power consumption, rapid thermal ramp times and four-color multiplex detection. Single tube multiplex assays can be used to normalize for RNA concentration and integrity, and to study multiple genes of interest in each module. The WetLab-2 system can downlink data from the ISS to the ground after a completed run and uplink new thermal cycling programs. The ability to conduct qRT-PCR and generate results on-orbit is an important step towards utilizing the ISS as a National Laboratory facility. Specifically, the ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. On orbit gene expression analysis can also eliminate 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 and provide on-orbit gene expression benchmarking prior to sample return. Finally, the system can also be used for analysis of

  4. Cloning and evaluation of reference genes for quantitative real-time PCR analysis in Amorphophallus

    Directory of Open Access Journals (Sweden)

    Kai Wang

    2017-04-01

    Full Text Available Quantitative real-time reverse transcription PCR (RT-qPCR has been widely used in the detection and quantification of gene expression levels because of its high accuracy, sensitivity, and reproducibility as well as its large dynamic range. However, the reliability and accuracy of RT-qPCR depends on accurate transcript normalization using stably expressed reference genes. Amorphophallus is a perennial plant with a high content of konjac glucomannan (KGM in its corm. This crop has been used as a food source and as a traditional medicine for thousands of years. Without adequate knowledge of gene expression profiles, there has been no report of validated reference genes in Amorphophallus. In this study, nine genes that are usually used as reference genes in other crops were selected as candidate reference genes. These putative sequences of these genes Amorphophallus were cloned by the use of degenerate primers. The expression stability of each gene was assessed in different tissues and under two abiotic stresses (heat and waterlogging in A. albus and A. konjac. Three distinct algorithms were used to evaluate the expression stability of the candidate reference genes. The results demonstrated that EF1-a, EIF4A, H3 and UBQ were the best reference genes under heat stress in Amorphophallus. Furthermore, EF1-a, EIF4A, TUB, and RP were the best reference genes in waterlogged conditions. By comparing different tissues from all samples, we determined that EF1-α, EIF4A, and CYP were stable in these sets. In addition, the suitability of these reference genes was confirmed by validating the expression of a gene encoding the small heat shock protein SHSP, which is related to heat stress in Amorphophallus. In sum, EF1-α and EIF4A were the two best reference genes for normalizing mRNA levels in different tissues and under various stress treatments, and we suggest using one of these genes in combination with 1 or 2 reference genes associated with different

  5. 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.

  6. 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

  7. 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...

  8. 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

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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)

  15. 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

  16. 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

  17. 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

  18. 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…

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 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.

  1. 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

  2. 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

  3. DEVELOPMENT OF AN ON-LINE, REAL-TIME ALPHA RADIATION MEASURING INSTRUMENT FOR LIQUID STREAMS

    International Nuclear Information System (INIS)

    1996-01-01

    The Department of Energy (DOE) has expressed a need for an on-line, real-time instrument for assaying alpha-emitting radionuclides (uranium and the transuranics) in effluent waters leaving DOE sites to ensure compliance with regulatory limits. Due to the short range of alpha particles in water (approximately40 Tm), it is necessary now to intermittently collect samples of water and send them to a central laboratory for analysis. A lengthy and costly procedure is used to separate and measure the radionuclides from each sample. Large variations in radionuclide concentrations in the water may go undetected due to the sporadic sampling. Even when detected, the reading may not be representative of the actual stream concentration. To address these issues, Tecogen, a division of Thermo Power Corporation, a Thermo Electron company, is developing a real-time, field-deployable, alpha monitor based on a solid-state silicon wafer semiconductor (patent pending, to be assigned to the Department of Energy). The Thermo Alpha Monitor (TAM) will serve to monitor effluent water streams (Subsurface Contaminants Focus Area) and will be suitable for process control of remediation as well as decontamination and decommissioning operations, such as monitoring scrubber or rinse water radioactivity levels (Mixed Waste Focus Area and D and D Focus Area). It would be applicable for assaying other liquids, such as oil, or solids after proper preconditioning. Rapid isotopic alpha air monitoring is also possible using this technology. This instrument for direct counting of alpha-emitters in aqueous streams is presently being developed by Thermo Power under a development program funded by the DOE Environmental Management program (DOE-EM), administered by the Morgantown Energy Technology Center (METC). Under this contract, Thermo Power has demonstrated a solid-state, silicon-based semiconductor instrument, which uses a proprietary film-based collection system to quantitatively extract the

  4. 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

  5. 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.

  6. 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.

  7. 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...

  8. 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

  9. 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

  10. 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.

  11. Effect of ionizing radiation on the quantitative detection of Salmonella using real-time PCR

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Sangyong; Jung, Jinwoo [Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Kim, Minjeong; Ryu, Sangryeol [Department of Food and Animal Biotechnology, School of Agricultural Biotechnology, Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921 (Korea, Republic of); Kim, Dongho [Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of)], E-mail: fungikim@kaeri.re.kr

    2008-09-15

    Food irradiation is an economically viable technology for inactivating foodborne pathogens, but irradiation can mask pathogens in unhygienically prepared food. The aim of this study was to investigate the effect of irradiation treatment on the detection of Salmonella using real-time PCR. Three commercially available kits were tested, of which the InstaGene Matrix procedure was most effective in preparing template DNA from Salmonella exposed to radiation in broth culture. The minimum level of detection by real-time PCR combined with InstaGene Matrix was 3 log units of Salmonella per milliliter. However, when pure cultures of Salmonella were irradiated at 3 and 5 kGy, the cycle threshold (C{sub T}) increased 1-1.5-fold compared to irradiation at 0 and 1 kGy. This indicated that irradiation treatment may result in an underestimation of bacterial counts due to radiation-induced DNA lesions. We also compared C{sub T} values in inoculated chicken homogenates before and after irradiation, which in this model caused a 1.3-3.3-fold underestimation of bacterial counts with respect to irradiation dose.

  12. Design and Validation of Real-Time PCR: Quantitative Diagnosis of Common Leishmania Species in Iran.

    Science.gov (United States)

    Fekri Soofi Abadi, Maryam; Dabiri, Shahriar; Fotouhi Ardakani, Reza; Fani Malaki, Lina; Amirpoor Rostami, Sahar; Ziasistani, Mahsa; Dabiri, Donya

    2016-07-01

    Design and validation of Real-time PCR on the protected gene region ITS2 to quantify the parasite load in common leishmania (L) species. Probe and primer were designed from the ITS2 region between the rRNA genes with minimum gene variation in three common leishmania species followed by a Real-time PCR using the Taq man probe method in the form of absolute quantification. A series of different concentrations of leishmania were analyzed. After the purified PCR product was successfully placed in a PTG19-T plasmid vector, specialized ITS2 region was cloned in this plasmid. In the last phase, the cloned gene was transferred to the Ecoli.Top10F bacteria. The standard plasmid was provided in 10(7) to 10(1) copies/rxn concentrations. The specification and clinical sensitivity of the data was analyzed using inter and intra scales. The probe and primer were designed using three species, including L. infantum, L. major, and L.tropica. Seven concentrations of purified parasite in culture media showed that the selected region for quantifying the parasite is suitable. Clinical and analytical specificity and sensitivity were both 100%, respectively. The Taq man method for the ITS2 region in leishmania is one the most sensitive diagnostic test for identifying the parasite load and is suggested as a tool for fast identification and quantification of species.

  13. 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.

  14. 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.

  15. 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

  16. 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...

  17. 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....

  18. 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)

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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)

  8. 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)

  9. 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.

  10. 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)

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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).

  17. Global real-time dose measurements using the Automated Radiation Measurements for Aerospace Safety (ARMAS) system

    Science.gov (United States)

    Tobiska, W. Kent; Bouwer, D.; Smart, D.; Shea, M.; Bailey, J.; Didkovsky, L.; Judge, K.; Garrett, H.; Atwell, W.; Gersey, B.; Wilkins, R.; Rice, D.; Schunk, R.; Bell, D.; Mertens, C.; Xu, X.; Wiltberger, M.; Wiley, S.; Teets, E.; Jones, B.; Hong, S.; Yoon, K.

    2016-11-01

    The Automated Radiation Measurements for Aerospace Safety (ARMAS) program has successfully deployed a fleet of six instruments measuring the ambient radiation environment at commercial aircraft altitudes. ARMAS transmits real-time data to the ground and provides quality, tissue-relevant ambient dose equivalent rates with 5 min latency for dose rates on 213 flights up to 17.3 km (56,700 ft). We show five cases from different aircraft; the source particles are dominated by galactic cosmic rays but include particle fluxes for minor radiation periods and geomagnetically disturbed conditions. The measurements from 2013 to 2016 do not cover a period of time to quantify galactic cosmic rays' dependence on solar cycle variation and their effect on aviation radiation. However, we report on small radiation "clouds" in specific magnetic latitude regions and note that active geomagnetic, variable space weather conditions may sufficiently modify the magnetospheric magnetic field that can enhance the radiation environment, particularly at high altitudes and middle to high latitudes. When there is no significant space weather, high-latitude flights produce a dose rate analogous to a chest X-ray every 12.5 h, every 25 h for midlatitudes, and every 100 h for equatorial latitudes at typical commercial flight altitudes of 37,000 ft ( 11 km). The dose rate doubles every 2 km altitude increase, suggesting a radiation event management strategy for pilots or air traffic control; i.e., where event-driven radiation regions can be identified, they can be treated like volcanic ash clouds to achieve radiation safety goals with slightly lower flight altitudes or more equatorial flight paths.

  18. 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

  19. 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

  20. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. Visualisation and quantitative analysis of the rodent malaria liver stage by real time imaging.

    NARCIS (Netherlands)

    Ploemen, I.H.J.; Prudencio, M.; Douradinha, B.G.; Ramesar, J.; Fonager, J.; Gemert, G.J.A. van; Luty, A.J.F.; Hermsen, C.C.; Sauerwein, R.W.; Baptista, F.G.; Mota, M.M.; Waters, A.P.; Que, I.; Lowik, C.W.G.M.; Khan, S.M.; Janse, C.J.; Franke-Fayard, B.

    2009-01-01

    The quantitative analysis of Plasmodium development in the liver in laboratory animals in cultured cells is hampered by low parasite infection rates and the complicated methods required to monitor intracellular development. As a consequence, this important phase of the parasite's life cycle has been

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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

  18. Quantitative Real-time Polymerase Chain Reaction for Enteropathogenic Escherichia coli: A Tool for Investigation of Asymptomatic Versus Symptomatic Infections

    Science.gov (United States)

    Barletta, Francesca; Mercado, Erik; Ruiz, Joaquim; Ecker, Lucie; Lopez, Giovanni; Mispireta, Monica; Gil, Ana I.; Lanata, Claudio F.; Cleary, Thomas G.

    2011-01-01

    Background. Enteropathogenic Escherichia coli (EPEC) strains are pediatric pathogens commonly isolated from both healthy and sick children with diarrhea in areas of endemicity. The aim of this study was to compare the bacterial load of EPEC isolated from stool samples from children with and without diarrhea to determine whether bacterial load might be a useful tool for further study of this phenomenon. Methods. EPEC was detected by polymerase chain reaction (PCR) of colonies isolated on MacConkey plates from 53 diarrheal and 90 healthy children aged <2 years. DNA was isolated from stool samples by cetyltrimethylammonium bromide extraction. To standardize quantification by quantitative real-time PCR (qRT-PCR), the correlation between fluorescence threshold cycle and copy number of the intimin gene of EPEC E2348/69 was determined. Results. The detection limit of qRT-PCR was 5 bacteria/mg stool. The geometric mean load in diarrhea was 299 bacteria/mg (95% confidence interval [CI], 77–1164 bacteria/mg), compared with 29 bacteria/mg (95% CI, 10–87 bacteria/mg) in control subjects (P = .016). Bacterial load was significantly higher in children with diarrhea than in control subjects among children <12 months of age (178 vs 5 bacteria/mg; P = .006) and among children with EPEC as the sole pathogen (463 vs 24 bacteria/mg; P = .006). Conclusions. EPEC load measured by qRT-PCR is higher in diarrheal than in healthy children. qRT-PCR may be useful to study the relationship between disease and colonization in settings of endemicity. PMID:22028433

  19. 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.

  20. 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)

  1. 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.

  2. 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

  3. 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.

  4. Comparison between quantitative nucleic acid sequence-based amplification, real-time reverse transcriptase PCR, and real-time PCR for quantification of Leishmania parasites

    NARCIS (Netherlands)

    van der Meide, Wendy; Guerra, Jorge; Schoone, Gerard; Farenhorst, Marit; Coelho, Leila; Faber, William; Peekel, Inge; Schallig, Henk

    2008-01-01

    DNA or RNA amplification methods for detection of Leishmania parasites have advantages regarding sensitivity and potential quantitative characteristics in comparison with conventional diagnostic methods but are often still not routinely applied. However, the use and application of molecular assays

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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.)

  13. Real-time quantitative Schlieren imaging by fast Fourier demodulation of a checkered backdrop

    Science.gov (United States)

    Wildeman, Sander

    2018-06-01

    A quantitative synthetic Schlieren imaging (SSI) method based on fast Fourier demodulation is presented. Instead of a random dot pattern (as usually employed in SSI), a 2D periodic pattern (such as a checkerboard) is used as a backdrop to the refractive object of interest. The range of validity and accuracy of this "Fast Checkerboard Demodulation" (FCD) method are assessed using both synthetic data and experimental recordings of patterns optically distorted by small waves on a water surface. It is found that the FCD method is at least as accurate as sophisticated, multi-stage, digital image correlation (DIC) or optical flow (OF) techniques used with random dot patterns, and it is significantly faster. Efficient, fully vectorized, implementations of both the FCD and DIC/OF schemes developed for this study are made available as open source Matlab scripts.

  14. 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

  15. 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.

  16. Measurement of gas phase characteristics using new monofiber optical probes and real time signal processing

    International Nuclear Information System (INIS)

    Cartellier, A.

    1998-01-01

    Single optical or impedance phase detection probes are able to measure gas velocities provided that their sensitive length L is accurately known. In this paper, it is shown that L can be controlled during the manufacture of optical probes. Beside, for a probe geometry in the form of a cone + a cylinder + a cone, the corresponding rise time / velocity correlation becomes weakly sensitive to uncontrollable parameter such as the angle of impact on the interface. A real time signal processing performing phase detection as well as velocity measurements is described. Since its sensitivity to the operator inputs is less than the reproducibility of measurements, it is a fairly objective tool. Qualifications achieved in air/water flows with various optical probes demonstrate that the void fraction is detected with a relative error less than 10 %. For bubbly flows, the gas flux is accurate within ±10%, but this uncertainty increases when large bubbles are present in the flow. (author)

  17. Phasor Measurement Unit and Phasor Data Concentrator test with Real Time Digital Simulator

    DEFF Research Database (Denmark)

    Diakos, Konstantinos; Wu, Qiuwei; Nielsen, Arne Hejde

    2014-01-01

    that is able to derive and communicate synchrophasor measurements of different parts of the power network and the development of tests, according to IEEE standards, that evaluate the performance of PMUs and PDCs. The tests are created by using a Real Time Digital Simulation (RTDS) system. The results obtained......The main focus of the electrical engineers nowadays, is to develop a smart grid that is able to monitor, evaluate and control the power system operation. The integration of Intelligent Electronic Devices (IED s) to the power network, is a strong indication of the inclination to lead the power...... network to a more reliable, secure and economic operation. The implementation of these devices though, demands the warranty of a secure operation and high-accuracy performance. This paper describes the procedure of establishing a PMU (Phasor Measurement Unit)–PDC (Phasor Data Concentrator) platform...

  18. 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.

  19. Basic evaluation of signal transmission in a real-time internal radiation dose measurement system

    International Nuclear Information System (INIS)

    Shinohe, K.; Takura, T.; Sato, F.; Matsuki, H.; Yamada, S.; Sato, T.

    2009-01-01

    In radiation therapy, excessive exposure to radiation occurs because the dose actually delivered to the tumor is not known. As a result, a patient suffers from side effects. To solve this problem, a system is needed in which the delivered dose is measured inside the body and the dose data are transmitted from inside to outside of the body during radiation therapy. If such a system is realized, it will be possible to treat cancer safely and effectively. The proposed real-time internal radiation dose measurement system consists of an implantable dosimeter, a wireless communication system, and a wireless feeding system. In this study, a wireless communication system that uses magnetic fields was investigated. As a result, a communication distance of 200 mm was obtained. It was confirmed that radiation dose data could be transmitted outside the body when the communication distance is the required 200 mm. (author)

  20. 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...

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. Visualisation of Radioactivity in Real-Time on a Tablet Measured by a Hybrid Pixel Detector

    CERN Document Server

    AUTHOR|(SzGeCERN)749233; Bantel, Michael; Grünhaupt, Ulrich

    This work explores a method to visualise and interact with radioactivity over time and space by means of augmented reality on a screen. A prototype, iPadPix, was built to demonstrate use as an intuitive new tool for educative and training purposes. Measured by a hybrid pixel detector, Timepix, traces of radioactive decays are displayed in real- time on a mobile device. Its detection principle and properties are detailed as well as the calibration of the sensor. An embedded board is used to process and forward the sensor data to a tablet over a wireless network connection. Software was developed to processes and overlay signatures of ionising radiation and particles on a live camera feed. It is described here and published as open source.

  7. 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”.

  8. 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.

  9. Real time management of the AD Schottky/BTF beam measurement system

    CERN Document Server

    Ludwig, M

    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.

  10. Real-time determination of fringe pattern frequencies: An application to pressure measurement

    Science.gov (United States)

    Sciammarella, Cesar A.; Piroozan, Parham

    2007-05-01

    Retrieving information in real time from fringe patterns is a topic of a great deal of interest in scientific and engineering applications of optical methods. This paper presents a method for fringe frequency determination based on the capability of neural networks to recognize signals that are similar but not identical to signals used to train the neural network. Sampled patterns are generated by calibration and stored in memory. Incoming patterns are analyzed by a back-propagation neural network at the speed of the recording device, a CCD camera. This method of information retrieval is utilized to measure pressures on a boundary layer flow. The sensor combines optics and electronics to analyze dynamic pressure distributions and to feed information to a control system that is capable to preserve the stability of the flow.

  11. 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.

  12. 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.

  13. Real-time measurements of endogenous CO production from vascular cells using an ultrasensitive laser sensor

    Science.gov (United States)

    Morimoto, Y.; Durante, W.; Lancaster, D. G.; Klattenhoff, J.; Tittel, F. K.

    2001-01-01

    Carbon monoxide (CO) has been implicated as a biological messenger molecule analogous to nitric oxide. A compact gas sensor based on a midinfrared laser absorption spectroscopy was developed for direct and real-time measurement of trace levels (in approximate pmol) of CO release by vascular cells. The midinfrared light is generated by difference frequency mixing of two nearinfrared lasers in a nonlinear optical crystal. A strong infrared absorption line of CO (4.61 microm) is chosen for convenient CO detection without interference from other gas species. The generation of CO from cultured vascular smooth muscle cells was detected every 20 s without any chemical modification to the CO. The sensitivity of the sensor reached 6.9 pmol CO. CO synthesis was measured from untreated control cells (0.25 nmol per 10(7) cells/h), sodium nitroprusside-treated cells (0.29 nmol per 10(7) cells/h), and hemin-treated cells (0.49 nmol per 10(7) cells/h). The sensor also detected decreases in CO production after the addition of the heme oxygenase (HO) inhibitor tin protoporphyrin-IX (from 0.49 to 0.02 nmol per 10(7) cells/h) and increases after the administration of the HO substrate hemin (from 0.27 to 0.64 nmol per 10(7) cells/h). These results demonstrate that midinfrared laser absorption spectroscopy is a useful technique for the noninvasive and real-time detection of trace levels of CO from biological tissues.

  14. 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.

  15. 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

  16. 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.

  17. Best Practice for Rainfall Measurement, Torrential Flood Monitoring and Real Time Alerting System in Serbia

    Science.gov (United States)

    Stefanovic, Milutin; Milojevic, Mileta; Zlatanovic, Nikola

    2014-05-01

    Serbia occupies 88.000 km2 and its confined zone menaced with torrent flood occupies 50.000km2. Floods on large rivers and torrents are the most frequent natural disasters in Serbia. This is the result of a geographic position and relief of Serbia. Therefore, defense from these natural disasters has been institutionalized since the 19th century. Through its specialized bodies and public companies, the State organized defense from floods on large rivers and protection of international and other main roads. The Topčiderska River is one of a number of rivers in Serbia that is a threat to both urban and rural environments. In this text, general characteristics of this river will be illustrated, as well as the historical natural hazards that have occurred in the part of Belgrade near Topčiderska River. Belgrade is the capital of Serbia, its political, administrative and financial center, which means that there are significant financial capacities and human resources for investments in all sectors, and specially in the water resources sector. Along the Topčiderska catchment there are many industrial, traffic and residential structures that are in danger of floods and flood protection is more difficult with rapid high flows. The goal is to use monitoring on the Topčiderska River basin to set up a modern system for monitoring in real time and forecast of torrential floods. This paper represents a system of remote detection and monitoring of torrential floods and rain measurements in real time on Topciderka river and ready for a quick response.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Real-Time Measurement of Width and Height of Weld Beads in GMAW Processes

    Directory of Open Access Journals (Sweden)

    Jesús Emilio Pinto-Lopera

    2016-09-01

    Full Text Available Associated to the weld quality, the weld bead geometry is one of the most important parameters in welding processes. It is a significant requirement in a welding project, especially in automatic welding systems where a specific width, height, or penetration of weld bead is needed. This paper presents a novel technique for real-time measuring of the width and height of weld beads in gas metal arc welding (GMAW using a single high-speed camera and a long-pass optical filter in a passive vision system. The measuring method is based on digital image processing techniques and the image calibration process is based on projective transformations. The measurement process takes less than 3 milliseconds per image, which allows a transfer rate of more than 300 frames per second. The proposed methodology can be used in any metal transfer mode of a gas metal arc welding process and does not have occlusion problems. The responses of the measurement system, presented here, are in a good agreement with off-line data collected by a common laser-based 3D scanner. Each measurement is compare using a statistical Welch’s t-test of the null hypothesis, which, in any case, does not exceed the threshold of significance level α = 0.01, validating the results and the performance of the proposed vision system.

  3. Real-time measurements of suspended sediment concentration and particle size using five techniques

    Science.gov (United States)

    Felix, D.; Albayrak, I.; Abgottspon, A.; Boes, R. M.

    2016-11-01

    Fine sediments are important in the design and operation of hydropower plants (HPPs), in particular with respect to sediment management and hydro-abrasive erosion in hydraulic machines. Therefore, there is a need for reliable real-time measurements of suspended sediment mass concentration (SSC) and particle size distribution (PSD). The following instruments for SSC measurements were investigated in a field study during several years at the HPP Fieschertal in the Swiss Alps: (1) turbidimeters, (2) a Laser In-Situ Scattering and Trans- missometry instrument (LISST), (3) a Coriolis Flow and Density Meter (CFDM), (4) acoustic transducers, and (5) pressure sensors. LISST provided PSDs in addition to concentrations. Reference SSCs were obtained by gravimetrical analysis of automatically taken water samples. In contrast to widely used turbidimeters and the single-frequency acoustic method, SSCs obtained from LISST, the CFDM or the pressure sensors were less or not affected by particle size variations. The CFDM and the pressure sensors allowed measuring higher SSC than the optical or the acoustic techniques (without dilution). The CFDM and the pressure sensors were found to be suitable to measure SSC ≥ 2 g/l. In this paper, the measuring techniques, instruments, setup, methods for data treatment, and selected results are presented and discussed.

  4. 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.

  5. 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

  6. 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.

  7. 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

  8. 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.

  9. Measurement system of correlation functions of microwave single photon source in real time

    Science.gov (United States)

    Korenkov, A.; Dmitriev, A.; Astafiev, O.

    2018-02-01

    Several quantum setups, such as quantum key distribution networks[1] and quantum simulators (e.g. boson sampling), by their design rely on single photon sources (SPSs). These quantum setups were demonstrated to operate in optical frequency domain. However, following the steady advances in circuit quantum electrodynamics, a proposal has been made recently[2] to demonstrate boson sampling with microwave photons. This in turn requires the development of reliable microwave SPS. It's one of the most important characteristics are the first-order and the second-order correlation functions g1 and g2. The measurement technique of g1 and g2 is significantly different from that in the optical domain [3],[4] because of the current unavailability of microwave single-photon detectors. In particular, due to high levels of noise present in the system a substantial amount of statistics in needed to be acquired. This work presents a platform for measurement of g1 and g2 that processes the incoming data in real time, maximizing the efficiency of data acquisition. The use of field-programmable gate array (FPGA) electronics, common in similar experiments[3] but complex in programming, is avoided; instead, the calculations are performed on a standard desktop computer. The platform is used to perform the measurements of the first-order and the second-order correlation functions of the microwave SPS.

  10. A cavity ring-down spectroscopy sensor for real-time Hall thruster erosion measurements

    International Nuclear Information System (INIS)

    Lee, B. C.; Huang, W.; Tao, L.; Yamamoto, N.; Yalin, A. P.; Gallimore, A. D.

    2014-01-01

    A continuous-wave cavity ring-down spectroscopy sensor for real-time measurements of sputtered boron from Hall thrusters has been developed. The sensor uses a continuous-wave frequency-quadrupled diode laser at 250 nm to probe ground state atomic boron sputtered from the boron nitride insulating channel. Validation results from a controlled setup using an ion beam and target showed good agreement with a simple finite-element model. Application of the sensor for measurements of two Hall thrusters, the H6 and SPT-70, is described. The H6 was tested at power levels ranging from 1.5 to 10 kW. Peak boron densities of 10 ± 2 × 10 14 m −3 were measured in the thruster plume, and the estimated eroded channel volume agreed within a factor of 2 of profilometry. The SPT-70 was tested at 600 and 660 W, yielding peak boron densities of 7.2 ± 1.1 × 10 14 m −3 , and the estimated erosion rate agreed within ∼20% of profilometry. Technical challenges associated with operating a high-finesse cavity in the presence of energetic plasma are also discussed

  11. Quantitative Determination of Plasmodium vivax Gametocytes by Real-Time Quantitative Nucleic Acid Sequence-Based Amplification in Clinical Samples

    NARCIS (Netherlands)

    Beurskens, Martijn; Mens, Pètra; Schallig, Henk; Syafruddin, Din; Asih, Puji Budi Setia; Hermsen, Rob; Sauerwein, Robert

    2009-01-01

    Microscopic detection of Plasmodium vivax gametocytes, the sexual life stage of this malaria parasite, is insensitive because P vivax parasitaemia is low. To detect and quantity gametocytes a more sensitive, quantitative realtime Pvs25-QT-NASBA based oil Pvs25 mRNA was developed and tested in two

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. [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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Two-phase fluid flow measurements in small diameter channels using real-time neutron radiography

    International Nuclear Information System (INIS)

    Carlisle, B.S.; Johns, R.C.; Hassan, Y.A.

    2004-01-01

    A series of real-time, neutron radiography, experiments are ongoing at the Texas A and M Nuclear Science Center Reactor (NSCR). These tests determine the resolving capabilities for radiographic imaging of two phase water and air flow regimes through small diameter flow channels. Though both film and video radiographic imaging is available, the real-time video imaging was selected to capture the dynamic flow patterns with results that continue to improve. (author)

  3. Measurement of density distribution of fluids by real-time holographic interferometer using lenticular lens

    International Nuclear Information System (INIS)

    Sakurai, Katsumi; Okamoto, Koji; Kato, Fumitake; Shimizu, Isao

    1998-01-01

    The three-dimensional density distribution could be measured using the computer tomography technique with interferogram image. The photoconductor-plastic hologram (PPH) is a new hologram device which can easily make the hologram for the real-time interferometer. Since the image contains 2D information, lots of images taken from different angles should be needed to reconstruct the 3D information. However, the optics configuration will be too complex to obtain the multi-directional image simultaneously, even in the PPH system. Using the diffusion plate, the laser light could be diffused to multi-direction. When the hologram is recorded with the diffused laser, the multi-directional image can be obtained using only one hologram plate. In the computer tomography technique, only the holizontal direction is effective, while the diffused laser contains the whole direction, causing the noise on the hologram. In this study, the lenticular lens is used as the diffusion plate. The lenticular lens reflects the laser only in horizontal direction without the vertical (non-horizontal) direction. Therefore the multi-directional fringe images could be clearly obtained. In the experiment the helium jet was measured to demonstrate the effectiveness of the proposed system. (author)

  4. 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.

  5. 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.

  6. 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.

  7. 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...

  8. DEVELOPMENT OF AN ON-LINE, REAL-TIME ALPHA RADIATION MEASURING INSTRUMENT FOR LIQUID STREAMS

    International Nuclear Information System (INIS)

    Unknown

    1999-01-01

    The US Department of Energy (DOE) has expressed a need for an on-line, real-time instrument for assaying alpha-emitting radionuclides (uranium and the transuranics) in effluent waters leaving DOE sites to ensure compliance with regulatory limits. Due to the short range of alpha particles in water (approximately40 Im), it is necessary now to intermittently collect samples of water and send them to a central laboratory for analysis. A lengthy and costly procedure is used to separate and measure the radionuclides from each sample. Large variations in radionuclide concentrations in the water may go undetected due to the sporadic sampling. Even when detected, the reading may not be representative of the actual stream concentration. To address these issues, the Advanced Technologies Group of Thermo Power Corporation (a Thermo Electron company) is developing a real-time, field-deployable alpha monitor based on a solid-state silicon wafer semiconductor (US Patent 5,652,013 and pending, assigned to the US Department of Energy). The Thermo Water Alpha Monitor will serve to monitor effluent water streams (Subsurface Contaminants Focus Area) and will be suitable for process control of remediation as well as decontamination and decommissioning (D and D) operations, such as monitoring scrubber or rinse water radioactivity levels (Mixed Waste, Plutonium, and D and D Focus Area). It would be applicable for assaying other liquids, such as oil, or solids after proper preconditioning. Rapid isotopic alpha air monitoring is also possible using this technology. This report details the program's accomplishments to date. Most significantly, the Alpha Monitoring Instrument was successfully field demonstrated on water 100X below the Environmental Protection Agency's proposed safe drinking water limit--down to under 1 pCi/1. During the Field Test, the Alpha Monitoring Instrument successfully analyzed isotopic uranium levels on a total of five different surface water, process water, and

  9. Environmental Measurement While Drilling System for Real-Time Field Screening of Contaminants

    International Nuclear Information System (INIS)

    Lockwood, G.J.; Normann, R.A.; Williams, C.V.

    1999-01-01

    Sampling during environmental drilling is essential to fully characterize the spatial distribution and migration of subsurface contaminants. However, analysis of the samples is expensive and time-consuming: off-site laboratory analysis can take weeks or months. Real-time information on environmental conditions, drill bit location and temperature during drilling is valuable in many environmental restoration operations. This type of information can be used to provide field screening data and improved efficiency of site characterization activities. The Environmental Measurement-While-Drilling (EMWD) System represents an innovative blending of new and existing technology in order to obtain real-time data during drilling. The system consists of two subsystems. The down-hole subsystem (at the drill bit) consists of sensors, a power supply, a signal conditioning and transmitter board, and a radio-frequency (RF) coaxial cable. The up-hole subsystem consists of a battery pack/coil, pickup coil, receiver, and personal computer. The system is compatible with fluid miser drill pipe, a directional drilling technique that uses minimal drilling fluids and generates little to no secondary waste. In EMWD, downhole sensors are located behind the drill bit and linked by a high-speed data transmission system to a computer at the surface. Sandia-developed Windowstrademark-based software is used for data display and storage. As drilling is conducted, data is collected on the nature and extent of contamination, enabling on-the-spot decisions regarding drilling and sampling strategies. Initially, the downhole sensor consisted of a simple gamma radiation detector, a Geiger-Mueller tube (GMT). The design includes data assurance techniques to increase safety by reducing the probability of giving a safe indication when an unsafe condition exists. The EMWD system has been improved by the integration of a Gamma Ray Spectrometer (GRS) in place of the GMT. The GRS consists of a sodium iodide

  10. Apple detection using infrared thermal image, 3: Real-time temperature measurement of apple tree

    International Nuclear Information System (INIS)

    Zhang, S.H.; Takahashi, T.; Fukuchi, H.; Sun, M.; Terao, H.

    1998-01-01

    In Part 1, we reported the thermal distribution characteristics and the identification methods of apples, leaves and branches by using the infrared thermal image at the specific time. This paper reports the temperature changing characteristics and the relationships among apples, leaves and air temperature based on the information measured by the infrared thermal image equipment in the real-time for 24 hours. As a result, it was confirmed that the average temperature of apples was 1 degree C or more higher than the one of the leaves, and the average temperature of the leaves was almost same as the air temperature within daytime and about 3 hours period after sunset. It was also clarified for a remarkable temperature difference not to exist for midnight and the early morning between the apples and the leaves, and both became almost as well as the air temperature. Moreover, a binary image was easily obtained and the apples could be detected by using this temperature difference informat

  11. KERMA-based radiation dose management system for real-time patient dose measurement

    Science.gov (United States)

    Kim, Kyo-Tae; Heo, Ye-Ji; Oh, Kyung-Min; Nam, Sang-Hee; Kang, Sang-Sik; Park, Ji-Koon; Song, Yong-Keun; Park, Sung-Kwang

    2016-07-01

    Because systems that reduce radiation exposure during diagnostic procedures must be developed, significant time and financial resources have been invested in constructing radiation dose management systems. In the present study, the characteristics of an existing ionization-based system were compared to those of a system based on the kinetic energy released per unit mass (KERMA). Furthermore, the feasibility of using the KERMA-based system for patient radiation dose management was verified. The ionization-based system corrected the effects resulting from radiation parameter perturbations in general radiography whereas the KERMA-based system did not. Because of this difference, the KERMA-based radiation dose management system might overestimate the patient's radiation dose due to changes in the radiation conditions. Therefore, if a correction factor describing the correlation between the systems is applied to resolve this issue, then a radiation dose management system can be developed that will enable real-time measurement of the patient's radiation exposure and acquisition of diagnostic images.

  12. The development of a versatile field program for measuring tritium in real-time

    International Nuclear Information System (INIS)

    Rego, J.H.; Smith, D.K.

    1994-04-01

    Robust sample handling and liquid scintillation counting procedures have been developed to routinely monitor tritium in the field relative to the 20,000 pCi/L drinking water standard. This procedure allows tritium to be monitored hourly during 24 hour drilling operations at depths in the saturated zone potentially contaminated by sub-surface nuclear weapons testing at the Nevada Test Site. Using retrofitted shock hardened and vibration damped counters and strict analytical protocols, tritium may be measured rapidly in the field under hostile conditions. Concentration standards and ''dead'' tritium backgrounds are prepared weekly in a central laboratory and delivered to remote monitoring locations where they are recounted daily as a check on counter efficiency and calibration. Portable counters are located in trailers and powered off a battery pack and line filter fed by mobile generator. Samples are typically ground-waters mixed with drilling fluids returned after circulation through a drill string. Fluids are aerated and ''de-foamed,'' filtered, mixed with scintillation cocktail and subsequently dark-adapted before counting. Besides meeting regulatory requirements, ''real-time'' monitoring affords drilling and field personnel maximum protection against potential exposure to radionuclides; for routine operations, tritium activities may not exceed a 10,000 pCi/L threshold

  13. A bio-inspired real-time capable artificial lateral line system for freestream flow measurements.

    Science.gov (United States)

    Abels, C; Qualtieri, A; De Vittorio, M; Megill, W M; Rizzi, F

    2016-06-03

    To enhance today's artificial flow sensing capabilities in aerial and underwater robotics, future robots could be equipped with a large number of miniaturized sensors distributed over the surface to provide high resolution measurement of the surrounding fluid flow. In this work we show a linear array of closely separated bio-inspired micro-electro-mechanical flow sensors whose sensing mechanism is based on a piezoresistive strain-gauge along a stress-driven cantilever beam, mimicking the biological superficial neuromasts found in the lateral line organ of fishes. Aiming to improve state-of-the-art flow sensing capability in autonomously flying and swimming robots, our artificial lateral line system was designed and developed to feature multi-parameter freestream flow measurements which provide information about (1) local flow velocities as measured by the signal amplitudes from the individual cantilevers as well as (2) propagation velocity, (3) linear forward/backward direction along the cantilever beam orientation and (4) periodicity of pulses or pulse trains determined by cross-correlating sensor signals. A real-time capable cross-correlation procedure was developed which makes it possible to extract freestream flow direction and velocity information from flow fluctuations. The computed flow velocities deviate from a commercial system by 0.09 m s(-1) at 0.5 m s(-1) and 0.15 m s(-1) at 1.0 m s(-1) flow velocity for a sampling rate of 240 Hz and a sensor distance of 38 mm. Although experiments were performed in air, the presented flow sensing system can be applied to underwater vehicles as well, once the sensors are embedded in a waterproof micro-electro-mechanical systems package.

  14. First measurement of pp neutrinos in real time in the Borexino detector

    Science.gov (United States)

    Mosteiro, Pablo

    2014-09-01

    The Sun is fueled by a series of nuclear reactions that produce the energy that makes it shine. Neutrinos (nu) produced by these nuclear reactions exit the Sun and reach Earth within minutes, providing us with key information about what goes on at the core of our star. For over twenty years since the first detection of solar neutrinos in the late 1960's, an apparent deficit in their detection rate was known as the Solar Neutrino Problem. Today, the Mikheyev-Smirnov-Wolfenstein (MSW) effect is the accepted mechanism by which neutrinos oscillate inside the Sun, arriving at Earth as a mixture of nue, numu and nutau, the latter two of which were invisible to early detectors. Several experiments have now confirmed the observation of neutrino oscillations. These experiments, when their results are combined together, have demonstrated that neutrino oscillations are well described by the Large Mixing Angle (LMA) solution of the MSW effect. This thesis presents the first measurement of pp neutrinos in the Borexino detector, which is another validation of the LMA-MSW model of neutrino oscillations. In addition, it is one more step towards the completion of the spectroscopy of pp chain neutrinos in Borexino, leaving only the extremely faint hep neutrinos undetected. This advance validates the experiment itself and its previous results. This is, furthermore, the first direct real-time measurement of pp neutrinos. We find a pp neutrino detection rate of 143+/-16 (stat)+/-10 (syst) cpd/100 t in the Borexino experiment, which translates, according to the LMA-MSW model, to (6.42+/-0.85)x1010 cm -2 s-1. We also report on a measurement of neutrons in a dedicated system within the Borexino detector, which resulted in an improved understanding of neutron rates in liquid scintillator detectors at Gran Sasso depths. This result is crucial to the development of novel direct dark matter detection experiments.

  15. CLEAR PM: Teaching, Outreach, and Research Through Real-Time Particulate Measurements

    Science.gov (United States)

    DeCarlo, P. F.

    2013-12-01

    An understanding of particulate matter (also called aerosols) can be made through measurement. This measurement does not change in value if it is made in a teaching, research, or outreach environment. A grant from the Camille and Henry Dreyfus Foundation provided funding to construct an instrument suite composed of 1-4 second measurements that are displayed in real-time through a software interface. This display module is called CLEAR PM (Chemistry Lessons Enabling Aerosol Realizations through Particulate Measurement), and was conceived to apply across outreach activities, teaching activities, and research activities. The construction and software design of CLEAR PM was done as part of a special topics course for chemistry and engineering graduate students at Drexel University. Measurement principles of the different (research grade) instruments were taught as part of the course, with emphasis put on the fundamental measurements and their limitations, and an introduction to data acquisition software was also integral to the teaching component. As a final project of the course graduate students were required to create a 'teaching' module that illustrates a chemistry or physics concept and utilizes the measurements of CLEAR PM. These modules ranged from gas-phase ozone chemistry creating secondary organic aerosols, to the wavelength dependent absorption profiles of wood smoke versus propane soot. The teaching modules developed by the graduate students have been used in outreach activities sponsored by The Franklin Institute and the Clean Air Council in Philadelphia, where underrepresented groups often make up a large fraction of the audience. CLEAR PM is designed to give students and citizens a hands-on opportunity to see how we measure and understand the world around us. As mentioned previously, the instruments that are part of CLEAR PM are research grade instruments, and are actively being used in research projects in the DeCarlo lab at Drexel to study particulate

  16. Reference gene selection for real-time quantitative PCR analysis of the mouse uterus in the peri-implantation period.

    Directory of Open Access Journals (Sweden)

    Pengfei Lin

    Full Text Available The study of uterine gene expression patterns is valuable for understanding the biological and molecular mechanisms that occur during embryo implantation. Real-time quantitative RT-PCR (qRT-PCR is an extremely sensitive technique that allows for the precise quantification of mRNA abundance; however, selecting stable reference genes suitable for the normalization of qRT-PCR data is required to avoid the misinterpretation of experimental results and erroneous analyses. This study employs several mouse models, including an early pregnancy, a pseudopregnancy, a delayed implantation and activation, an artificial decidualization and a hormonal treatment model; ten candidate reference genes (PPIA, RPLP0, HPRT1, GAPDH, ACTB, TBP, B2M, 18S, UBC and TUBA that are found in uterine tissues were assessed for their suitability as internal controls for relative qRT-PCR quantification. GeNorm(PLUS, NormFinder, and BestKeeper were used to evaluate these candidate reference genes, and all of these methods identified RPLP0 and GAPDH as the most stable candidates and B2M and 18S as the least stable candidates. However, when the different models were analyzed separately, the reference genes exhibited some variation in their expression levels.

  17. Comparison of commercial DNA preparation kits for the detection of Brucellae in tissue using quantitative real-time PCR

    Directory of Open Access Journals (Sweden)

    Straube Eberhard

    2010-04-01

    Full Text Available Abstract Background The detection of Brucellae in tissue specimens using PCR assays is difficult because the amount of bacteria is usually low. Therefore, optimised DNA extraction methods are critical. The aim of this study was to assess the performance of commercial kits for the extraction of Brucella DNA. Methods Five kits were evaluated using clinical specimens: QIAamp™ DNA Mini Kit (QIAGEN, peqGold™ Tissue DNA Mini Kit (PeqLab, UltraClean™ Tissue and Cells DNA Isolation Kit (MoBio, DNA Isolation Kit for Cells and Tissues (Roche, and NucleoSpin™ Tissue (Macherey-Nagel. DNA yield was determined using a quantitative real-time PCR assay targeting IS711 that included an internal amplification control. Results Kits of QIAGEN and Roche provided the highest amount of DNA, Macherey-Nagel and Peqlab products were intermediate whereas MoBio yielded the lowest amount of DNA. Differences were significant (p Conclusions We observed differences in DNA yield as high as two orders of magnitude for some samples between the best and the worst DNA extraction kits and inhibition was observed occasionally. This indicates that DNA purification may be more relevant than expected when the amount of DNA in tissue is very low.

  18. Effectiveness of quantitative real time PCR in long-term follow-up of chronic myeloid leukemia patients

    International Nuclear Information System (INIS)

    Savasoglu, K.; Berber, B.

    2015-01-01

    To determine the use of the Quantitative Real Time PCR (RQ-PCR) assay follow-up with Chronic Myeloid Leukemia (CML) patients. Study Design: Cross-sectional observational. Place and Duration of Study: Izmir Ataturk Education and Research Hospital, Izmir, Turkey, from 2009 to 2013. Methodology: Cytogenetic, FISH, RQ-PCR test results from 177 CML patients 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. Results: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 < 0.001). RQ-PCR test failure rate did not correlate with other two tests (p > 0.05); however, karyotyping and FISH test failure rate was statistically significant (p < 0.001). Conclusion: Besides, the situation needed for karyotype analysis, RQ-PCR assay can be used alone in the follow-up of CML disease. (author)

  19. Quantitative real-time PCR technique for the identification of E. coli residual DNA in streptokinase recombinant product.

    Science.gov (United States)

    Fazelahi, Mansoureh; Kia, Vahid; Kaghazian, Hooman; Paryan, Mahdi

    2017-11-26

    Recombinant streptokinase is a biopharmaceutical which is usually produced in E. coli. Residual DNA as a contamination and risk factor may remain in the product. It is necessary to control the production procedure to exclude any possible contamination. The aim of the present study was to develop a highly specific and sensitive quantitative real-time PCR-based method to determine the amount of E. coli DNA in recombinant streptokinase. A specific primers and a probe was designed to detect all strains of E. coli. To determine the specificity, in addition to using NCBI BLASTn, 28 samples including human, bacterial, and viral genomes were used. The results confirmed that the assay detects no genomic DNA but E. coli's and the specificity was determined to be 100%. To determine the sensitivity and limit of detection of the assay, a 10-fold serial dilution (10 1 to 10 7 copies/µL) was tested in triplicate. The sensitivity of the test was determined to be 101 copies/µL or 35 fg/µL. Inter-assay and intra-assay were determined to be 0.86 and 1.69%, respectively. Based on the results, this assay can be used as an accurate method to evaluate the contamination of recombinant streptokinase in E. coli.

  20. Identification of reference genes for expression analysis by real-time quantitative PCR in drought-stressed soybean

    Directory of Open Access Journals (Sweden)

    Renata Stolf-Moreira

    2011-01-01

    Full Text Available The objective of this work was to validate, by quantitative PCR in real time (RT-qPCR, genes to be used as reference in studies of gene expression in soybean in drought-stressed trials. Four genes commonly used in soybean were evaluated: Gmβ-actin, GmGAPDH, GmLectin and GmRNAr18S. Total RNA was extracted from six samples: three from roots in a hydroponic system with different drought intensities (0, 25, 50, 75 and 100 minutes of water stress, and three from leaves of plants grown in sand with different soil moistures (15, 5 and 2.5% gravimetric humidity. The raw cycle threshold (Ct data were analyzed, and the efficiency of each primer was calculated for an overall analysis of the Ct range among the different samples. The GeNorm application was used to evaluate the best reference gene, according to its stability. The GmGAPDH was the least stable gene, with the highest mean values of expression stability (M, and the most stable genes, with the lowest M values, were the Gmβ-actin and GmRNAr18S, when both root and leaves samples were tested. These genes can be used in RT-qPCR as reference gene for expression analysis.

  1. 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.

  2. Development of real-time quantitative polymerase chain reaction assays to track treatment response in retinoid resistant acute promyelocytic leukemia

    Directory of Open Access Journals (Sweden)

    Jelena V Jovanovic

    2011-10-01

    Full Text Available Molecular detection of minimal residual disease (MRD has become established to assess remission status and guide therapy in patients with PML-RARA+ acute promyelocytic leukemia (APL. However, there are few data on tracking disease response in patients with rarer retinoid resistant subtypes of APL, characterized by PLZF-RARA and STAT5b-RARA. Despite their relative rarity (<1% of APL we identified 6 cases (PLZF-RARA, n=5; STAT5b-RARA, n=1, established the respective breakpoint junction regions and designed real-time quantitative polymerase chain reaction (RQ-PCR assays to detect leukemic transcripts. The relative level of fusion gene expression in diagnostic samples was comparable to that observed in t(15;17-associated APL, affording assay sensitivities of ~1 in 104-105. Serial samples were available from 2 PLZF-RARA APL patients. One showed persistent PCR positivity, predicting subsequent relapse, and remains in CR2, ~11 years post-autograft. The other, achieved molecular remission (CRm with combination chemotherapy, remaining in CR1 at 6 years. The STAT5b-RARA patient failed to achieve CRm following frontline combination chemotherapy and ultimately proceeded to allogeneic transplant on the basis of a steadily rising fusion transcript level. These data highlight the potential of RQ-PCR detection of MRD to facilitate development of more individualized approaches to the management of rarer molecularly-defined subsets of acute leukemia.

  3. Identification and validation of reference genes for quantitative real-time PCR in Drosophila suzukii (Diptera: Drosophilidae.

    Directory of Open Access Journals (Sweden)

    Yifan Zhai

    Full Text Available To accurately evaluate gene expression levels and obtain more accurate quantitative real-time RT-PCR (qRT-PCR data, normalization relative to reliable reference gene(s is required. Drosophila suzukii, is an invasive fruit pest native to East Asia, and recently invaded Europe and North America, the stability of its reference genes have not been previously investigated. In this study, ten candidate reference genes (RPL18, RPS3, AK, EF-1β, TBP, NADH, HSP22, GAPDH, Actin, α-Tubulin, were evaluated for their suitability as normalization genes under different biotic (developmental stage, tissue and population, and abiotic (photoperiod, temperature conditions. The three statistical approaches (geNorm, NormFinder and BestKeeper and one web-based comprehensive tool (RefFinder were used to normalize analysis of the ten candidate reference genes identified α-Tubulin, TBP and AK as the most stable candidates, while HSP22 and Actin showed the lowest expression stability. We used three most stable genes (α-Tubulin, TBP and AK and one unstably expressed gene to analyze the expression of P-glycoprotein in abamectin-resistant and sensitive strains, and the results were similar to reference genes α-Tubulin, TBP and AK, which show good stability, while the result of HSP22 has a certain bias. The three validated reference genes can be widely used for quantification of target gene expression with qRT-PCR technology in D.suzukii.

  4. Accurate, Fast and Cost-Effective Diagnostic Test for Monosomy 1p36 Using Real-Time Quantitative PCR

    Directory of Open Access Journals (Sweden)

    Pricila da Silva Cunha

    2014-01-01

    Full Text Available Monosomy 1p36 is considered the most common subtelomeric deletion syndrome in humans and it accounts for 0.5–0.7% of all the cases of idiopathic intellectual disability. The molecular diagnosis is often made by microarray-based comparative genomic hybridization (aCGH, which has the drawback of being a high-cost technique. However, patients with classic monosomy 1p36 share some typical clinical characteristics that, together with its common prevalence, justify the development of a less expensive, targeted diagnostic method. In this study, we developed a simple, rapid, and inexpensive real-time quantitative PCR (qPCR assay for targeted diagnosis of monosomy 1p36, easily accessible for low-budget laboratories in developing countries. For this, we have chosen two target genes which are deleted in the majority of patients with monosomy 1p36: PRKCZ and SKI. In total, 39 patients previously diagnosed with monosomy 1p36 by aCGH, fluorescent in situ hybridization (FISH, and/or multiplex ligation-dependent probe amplification (MLPA all tested positive on our qPCR assay. By simultaneously using these two genes we have been able to detect 1p36 deletions with 100% sensitivity and 100% specificity. We conclude that qPCR of PRKCZ and SKI is a fast and accurate diagnostic test for monosomy 1p36, costing less than 10 US dollars in reagent costs.

  5. 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.

  6. Identification of Reference Genes for Quantitative Real Time PCR Assays in Aortic Tissue of Syrian Hamsters with Bicuspid Aortic Valve.

    Science.gov (United States)

    Rueda-Martínez, Carmen; Fernández, M Carmen; Soto-Navarrete, María Teresa; Jiménez-Navarro, Manuel; Durán, Ana Carmen; Fernández, Borja

    2016-01-01

    Bicuspid aortic valve (BAV) is the most frequent congenital cardiac malformation in humans, and appears frequently associated with dilatation of the ascending aorta. This association is likely the result of a common aetiology. Currently, a Syrian hamster strain with a relatively high (∼40%) incidence of BAV constitutes the only spontaneous animal model of BAV disease. The characterization of molecular alterations in the aorta of hamsters with BAV may serve to identify pathophysiological mechanisms and molecular markers of disease in humans. In this report, we evaluate the expression of ten candidate reference genes in aortic tissue of hamsters in order to identify housekeeping genes for normalization using quantitative real time PCR (RT-qPCR) assays. A total of 51 adult (180-240 days old) and 56 old (300-440 days old) animals were used. They belonged to a control strain of hamsters with normal, tricuspid aortic valve (TAV; n = 30), or to the affected strain of hamsters with TAV (n = 45) or BAV (n = 32). The expression stability of the candidate reference genes was determined by RT-qPCR using three statistical algorithms, GeNorm, NormFinder and Bestkeeper. The expression analyses showed that the most stable reference genes for the three algorithms employed were Cdkn1β, G3pdh and Polr2a. We propose the use of Cdkn1β, or both Cdkn1β and G3pdh as reference genes for mRNA expression analyses in Syrian hamster aorta.

  7. Identification of Reference Genes for Quantitative Real Time PCR Assays in Aortic Tissue of Syrian Hamsters with Bicuspid Aortic Valve.

    Directory of Open Access Journals (Sweden)

    Carmen Rueda-Martínez

    Full Text Available Bicuspid aortic valve (BAV is the most frequent congenital cardiac malformation in humans, and appears frequently associated with dilatation of the ascending aorta. This association is likely the result of a common aetiology. Currently, a Syrian hamster strain with a relatively high (∼40% incidence of BAV constitutes the only spontaneous animal model of BAV disease. The characterization of molecular alterations in the aorta of hamsters with BAV may serve to identify pathophysiological mechanisms and molecular markers of disease in humans. In this report, we evaluate the expression of ten candidate reference genes in aortic tissue of hamsters in order to identify housekeeping genes for normalization using quantitative real time PCR (RT-qPCR assays. A total of 51 adult (180-240 days old and 56 old (300-440 days old animals were used. They belonged to a control strain of hamsters with normal, tricuspid aortic valve (TAV; n = 30, or to the affected strain of hamsters with TAV (n = 45 or BAV (n = 32. The expression stability of the candidate reference genes was determined by RT-qPCR using three statistical algorithms, GeNorm, NormFinder and Bestkeeper. The expression analyses showed that the most stable reference genes for the three algorithms employed were Cdkn1β, G3pdh and Polr2a. We propose the use of Cdkn1β, or both Cdkn1β and G3pdh as reference genes for mRNA expression analyses in Syrian hamster aorta.

  8. Standardisation and evaluation of a quantitative multiplex real-time PCR assay for the rapid identification of Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Feroze Ahmed Ganaie

    2015-01-01

    Full Text Available Rapid diagnosis of Streptococcus pneumoniae can play a significant role in decreasing morbidity and mortality of infection. The accurate diagnosis of pneumococcal disease is hampered by the difficulties in growing the isolates from clinical specimens and also by misidentification. Molecular methods have gained popularity as they offer improvement in the detection of causative pathogens with speed and ease. The present study aims at validating and standardising the use of 4 oligonucleotide primer-probe sets (pneumolysin [ply], autolysin [lytA], pneumococcal surface adhesion A [psaA] and Spn9802 [DNA fragment] in a single-reaction mixture for the detection and discrimination of S. pneumoniae. Here, we validate a quantitative multiplex real-time PCR (qmPCR assay with a panel consisting of 43 S. pneumoniae and 29 non-pneumococcal isolates, 20 culture positive, 26 culture negative and 30 spiked serum samples. A standard curve was obtained using S. pneumoniae ATCC 49619 strain and glyceraldehyde 3-phosphate dehydrogenase (GAPDH gene was used as an endogenous internal control. The experiment showed high sensitivity with lower limit of detection equivalent to 4 genome copies/µl. The efficiency of the reaction was 100% for ply, lytA, Spn9802 and 97% for psaA. The test showed sensitivity and specificity of 100% with culture isolates and serum specimens. This study demonstrates that qmPCR analysis of sera using 4 oligonucleotide primers appears to be an appropriate method for the genotypic identification of S. pneumoniae infection.

  9. Selection of reliable reference genes for quantitative real-time PCR in human T cells and neutrophils

    Directory of Open Access Journals (Sweden)

    Ledderose Carola

    2011-10-01

    Full Text Available Abstract Background The choice of reliable reference genes is a prerequisite for valid results when analyzing gene expression with real-time quantitative PCR (qPCR. This method is frequently applied to study gene expression patterns in immune cells, yet a thorough validation of potential reference genes is still lacking for most leukocyte subtypes and most models of their in vitro stimulation. In the current study, we evaluated the expression stability of common reference genes in two widely used cell culture models-anti-CD3/CD28 activated T cells and lipopolysaccharide stimulated neutrophils-as well as in unselected untreated leukocytes. Results The mRNA expression of 17 (T cells, 7 (neutrophils or 8 (unselected leukocytes potential reference genes was quantified by reverse transcription qPCR, and a ranking of the preselected candidate genes according to their expression stability was calculated using the programs NormFinder, geNorm and BestKeeper. IPO8, RPL13A, TBP and SDHA were identified as suitable reference genes in T cells. TBP, ACTB and SDHA were stably expressed in neutrophils. TBP and SDHA were also the most stable genes in untreated total blood leukocytes. The critical impact of reference gene selection on the estimated target gene expression is demonstrated for IL-2 and FIH expression in T cells. Conclusions The study provides a shortlist of suitable reference genes for normalization of gene expression data in unstimulated and stimulated T cells, unstimulated and stimulated neutrophils and in unselected leukocytes.

  10. Development of Real-Time Quantitative Polymerase Chain Reaction Assays to Track Treatment Response in Retinoid Resistant Acute Promyelocytic Leukemia

    Energy Technology Data Exchange (ETDEWEB)

    Jovanovic, Jelena V. [Cancer Genetics Laboratory, Department of Medical and Molecular Genetics, King’s College London School of Medicine, London (United Kingdom); Rennie, Kristian [GSTS Pathology, Guy’s Hospital, London (United Kingdom); Culligan, Dominic [Department of Haematology, Aberdeen Royal Infirmary, Aberdeen (United Kingdom); Peniket, Andrew [Department of Haematology, John Radcliffe Hospital, Oxford (United Kingdom); Lennard, Anne [Department of Haematology, Royal Victoria Infirmary, Newcastle (United Kingdom); Harrison, Justin [Department of Haematology, Hemel Hempstead Hospital, Hemel Hempstead (United Kingdom); Vyas, Paresh [Medical Research Council Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Oxford (United Kingdom); Grimwade, David, E-mail: david.grimwade@genetics.kcl.ac.uk [Cancer Genetics Laboratory, Department of Medical and Molecular Genetics, King’s College London School of Medicine, London (United Kingdom)

    2011-10-25

    Molecular detection of minimal residual disease (MRD) has become established to assess remission status and guide therapy in patients with ProMyelocytic Leukemia–RARA+ acute promyelocytic leukemia (APL). However, there are few data on tracking disease response in patients with rarer retinoid resistant subtypes of APL, characterized by PLZF–RARA and STAT5b–RARA. Despite their rarity (<1% of APL) we identified 6 cases (PLZF–RARA, n = 5; STAT5b–RARA, n = 1), established the respective breakpoint junction regions and designed reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) assays to detect leukemic transcripts. The relative level of fusion gene expression in diagnostic samples was comparable to that observed in t(15;17) – associated APL, affording assay sensitivities of ∼1 in 10{sup 4}−10{sup 5}. Serial samples were available from two PLZF–RARA APL patients. One showed persistent polymerase chain reaction positivity, predicting subsequent relapse, and remains in CR2, ∼11 years post-autograft. The other, achieved molecular remission (CRm) with combination chemotherapy, remaining in CR1 at 6 years. The STAT5b–RARA patient failed to achieve CRm following frontline combination chemotherapy and ultimately proceeded to allogeneic transplant on the basis of a steadily rising fusion transcript level. These data highlight the potential of RT-qPCR detection of MRD to facilitate development of more individualized approaches to the management of rarer molecularly defined subsets of acute leukemia.

  11. Development of Real-Time Quantitative Polymerase Chain Reaction Assays to Track Treatment Response in Retinoid Resistant Acute Promyelocytic Leukemia

    International Nuclear Information System (INIS)

    Jovanovic, Jelena V.; Rennie, Kristian; Culligan, Dominic; Peniket, Andrew; Lennard, Anne; Harrison, Justin; Vyas, Paresh; Grimwade, David

    2011-01-01

    Molecular detection of minimal residual disease (MRD) has become established to assess remission status and guide therapy in patients with ProMyelocytic Leukemia–RARA+ acute promyelocytic leukemia (APL). However, there are few data on tracking disease response in patients with rarer retinoid resistant subtypes of APL, characterized by PLZF–RARA and STAT5b–RARA. Despite their rarity (<1% of APL) we identified 6 cases (PLZF–RARA, n = 5; STAT5b–RARA, n = 1), established the respective breakpoint junction regions and designed reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) assays to detect leukemic transcripts. The relative level of fusion gene expression in diagnostic samples was comparable to that observed in t(15;17) – associated APL, affording assay sensitivities of ∼1 in 10 4 −10 5 . Serial samples were available from two PLZF–RARA APL patients. One showed persistent polymerase chain reaction positivity, predicting subsequent relapse, and remains in CR2, ∼11 years post-autograft. The other, achieved molecular remission (CRm) with combination chemotherapy, remaining in CR1 at 6 years. The STAT5b–RARA patient failed to achieve CRm following frontline combination chemotherapy and ultimately proceeded to allogeneic transplant on the basis of a steadily rising fusion transcript level. These data highlight the potential of RT-qPCR detection of MRD to facilitate development of more individualized approaches to the management of rarer molecularly defined subsets of acute leukemia.

  12. Chemical luminescence measurement of singlet oxygen generated by photodynamic therapy in solutions in real time

    Science.gov (United States)

    Luo, Shiming; Xing, Da; Zhou, Jing; Qin, Yanfang; Chen, Qun

    2005-04-01

    Photodynamic therapy (PDT) is a cancer therapy that utilizes optical energy to activate a photosensitizer drug in a target tissue. Reactive oxygen species (ROS), such as 1O2 and superoxide, are believed to be the major cytotoxic agents involved in PDT. Although current PDT dosimetry mostly involves measurements of light and photosensitizer doses delivered to a patient, the quantification of ROS production during a treatment would be the ultimate dosimetry of PDT. Technically, it is very difficult and expensive to directly measure the fluorescence from 1O2, due to its extreme short lifetime and weak signal strength. In this paper, Photofrin(R) and 635nm laser were used to generate 1O2 and superoxide in a PDT in solution. Compound 3,7- dihydro-6-{4-[2-(N"-(5-fluoresceinyl) thioureido) ethoxy] phenyl}-2- methylimidazo{1,2-a} pyrazin-3-one sodium salt,an Cyp- ridina luciferin analog commonly referred as FCLA, was used as a chemical reporter of ROS. The 532nm chemiluminescence (CL) from the reaction of the FCLA and ROS was detected with a photon multiplier tube (PMT) system operating at single photon counting mode. With the setup, we have made detections of ROS generated by PDT in real time. By varying the amount of conventional PDT dosage (photosensitizer concentration, light irradiation fluence and its delivery rate) and the amount of FCLA, the intensity of CL and its consumption rate were investigated. The results show that the intensity and temporal profile of CL are highly related to the PDT treatment parameters. This suggests that FCLA CL may provide a highly potential alternative for ROS detection during PDT.

  13. Real-time emission factor measurements of isocyanic acid from light duty gasoline vehicles.

    Science.gov (United States)

    Brady, James M; Crisp, Timia A; Collier, Sonya; Kuwayama, Toshihiro; Forestieri, Sara D; Perraud, Véronique; Zhang, Qi; Kleeman, Michael J; Cappa, Christopher D; Bertram, Timothy H

    2014-10-07

    Exposure to gas-phase isocyanic acid (HNCO) has been previously shown to be associated with the development of atherosclerosis, cataracts and rheumatoid arthritis. As such, accurate emission inventories for HNCO are critical for modeling the spatial and temporal distribution of HNCO on a regional and global scale. To date, HNCO emission rates from light duty gasoline vehicles, operated under driving conditions, have not been determined. Here, we present the first measurements of real-time emission factors of isocyanic acid from a fleet of eight light duty gasoline-powered vehicles (LDGVs) tested on a chassis dynamometer using the Unified Driving Cycle (UC) at the California Air Resources Board (CARB) Haagen-Smit test facility, all of which were equipped with three-way catalytic converters. HNCO emissions were observed from all vehicles, in contrast to the idealized laboratory measurements. We report the tested fleet averaged HNCO emission factors, which depend strongly on the phase of the drive cycle; ranging from 0.46 ± 0.13 mg kg fuel(-1) during engine start to 1.70 ± 1.77 mg kg fuel(-1) during hard acceleration after the engine and catalytic converter were warm. The tested eight-car fleet average fuel based HNCO emission factor was 0.91 ± 0.58 mg kg fuel(-1), within the range previously estimated for light duty diesel-powered vehicles (0.21-3.96 mg kg fuel(-1)). Our results suggest that HNCO emissions from LDGVs represent a significant emission source in urban areas that should be accounted for in global and regional models.

  14. 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...

  15. 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.

  16. 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

  17. Real-time measurement of outdoor worker's exposure to solar ultraviolet radiation in Pretoria, South Africa

    Directory of Open Access Journals (Sweden)

    Mmathapelo Makgabutlane

    2015-05-01

    Full Text Available The city of Pretoria in South Africa receives considerable solar ultraviolet radiation (UVR because of its low latitude (22–35°S and relatively clear skies. Certain meteorological factors affect the amount of solar UVR that reaches the ground; the most dominant factors being stratospheric ozone, cloud cover and solar zenith angle. It is known that overexposure to solar UVR may lead to the development of adverse health conditions, the most significant being skin cancer. Outdoor workers spend a significant amount of time outside and are thus susceptible to this risk. In this case study, we estimated, for the first time, the real-time solar UVR exposure of an outdoor worker in Pretoria. Measurements were made on 27 and 28 May 2013 using a handheld ultraviolet index (UVI meter calibrated against a science-grade biometer at the South African Weather Service in Pretoria. Personal exposure estimation was used to discern the pattern in diurnal and annual sunburn risk for the outdoor worker. Ambient UVR levels ranged from 0 UVI to 4.66 UVI and the outdoor worker’s potential exposure estimates regularly exceeded 80% of these levels depending on the time of day. The risk of sunburn was evident; however, actual incidents would depend on individual skin photosensitivity and melanin content, as well as sun protection used. Further research is needed to determine the personal exposure estimations of outdoor workers in other provinces in which solar UVR levels may be equally high, or higher than those in Pretoria.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.

    Directory of Open Access Journals (Sweden)

    Ruby Chandna

    Full Text Available The real time quantitative reverse transcription PCR (qRT-PCR is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes, ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes, in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.

  5. SNPs and real-time quantitative PCR method for constitutional allelic copy number determination, the VPREB1 marker case

    Directory of Open Access Journals (Sweden)

    Costa Elena

    2011-05-01

    Full Text Available Abstract Background 22q11.2 microdeletion is responsible for the DiGeorge Syndrome, characterized by heart defects, psychiatric disorders, endocrine and immune alterations and a 1 in 4000 live birth prevalence. Real-time quantitative PCR (qPCR approaches for allelic copy number determination have recently been investigated in 22q11.2 microdeletions detection. The qPCR method was performed for 22q11.2 microdeletions detection as a first-level screening approach in a genetically unknown series of patients with congenital heart defects. A technical issue related to the VPREB1 qPCR marker was pointed out. Methods A set of 100 unrelated Italian patients with congenital heart defects were tested for 22q11.2 microdeletions by a qPCR method using six different markers. Fluorescence In Situ Hybridization technique (FISH was used for confirmation. Results qPCR identified six patients harbouring the 22q11.2 microdeletion, confirmed by FISH. The VPREB1 gene marker presented with a pattern consistent with hemideletion in one 3 Mb deleted patient, suggestive for a long distal deletion, and in additional five non-deleted patients. The long distal 22q11.2 deletion was not confirmed by Comparative Genomic Hybridization. Indeed, the VPREB1 gene marker generated false positive results in association with the rs1320 G/A SNP, a polymorphism localized within the VPREB1 marker reverse primer sequence. Patients heterozygous for rs1320 SNP, showed a qPCR profile consistent with the presence of a hemideletion. Conclusions Though the qPCR technique showed advantages as a screening approach in terms of cost and time, the VPREB1 marker case revealed that single nucleotide polymorphisms can interfere with qPCR data generating erroneous allelic copy number interpretations.

  6. Selection and validation of appropriate reference genes for quantitative real-time PCR analysis in Salvia hispanica.

    Directory of Open Access Journals (Sweden)

    Rahul Gopalam

    Full Text Available Quantitative real-time polymerase chain reaction (qRT-PCR has become the most popular choice for gene expression studies. For accurate expression analysis, it is pertinent to select a stable reference gene to normalize the data. It is now known that the expression of internal reference genes varies considerably during developmental stages and under different experimental conditions. For Salvia hispanica, an economically important oilseed crop, there are no reports of stable reference genes till date. In this study, we chose 13 candidate reference genes viz. Actin11 (ACT, Elongation factor 1-alpha (EF1-α, Eukaryotic translation initiation factor 3E (ETIF3E, alpha tubulin (α-TUB, beta tubulin (β-TUB, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Cyclophilin (CYP, Clathrin adaptor complex (CAC, Serine/threonine-protein phosphatase 2A (PP2A, FtsH protease (FtsH, 18S ribosomal RNA (18S rRNA, S-adenosyl methionine decarboxylase (SAMDC and Rubisco activase (RCA and the expression levels of these genes were assessed in a diverse set of tissue samples representing vegetative stages, reproductive stages and various abiotic stress treatments. Two of the widely used softwares, geNorm and Normfinder were used to evaluate the expression stabilities of these 13 candidate reference genes under different conditions. Results showed that GAPDH and CYP expression remain stable throughout in the different abiotic stress treatments, CAC and PP2A expression were relatively stable under reproductive stages and α-TUB, PP2A and ETIF3E were found to be stably expressed in vegetative stages. Further, the expression levels of Diacylglycerol acyltransferase (DGAT1, a key enzyme in triacylglycerol synthesis was analyzed to confirm the validity of reference genes identified in the study. This is the first systematic study of selection of reference genes in S. hispanica, and will benefit future expression studies in this crop.

  7. Validation of reference genes for quantitative expression analysis by real-time rt-PCR in four lepidopteran insects.

    Science.gov (United States)

    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), Chilo suppressalis Walker (Crambidae), and Spodoptera exigua Hübner (Noctuidae) to study their expression stability. The algorithms of geNorm, NormFinder, stability index, and ΔCt analysis were used to evaluate these HKGs. Across different developmental stages, actin A1 was the most stable in P. xylostella and C. suppressalis, but it was the least stable in B. mori and S. exigua. Rp49 and GAPDH were the most stable in B. mori and S. exigua, respectively. In different tissues, GAPDH, E2F, and Rp49 were the most stable in B. mori, S. exigua, and C. suppressalis, respectively. The relative abundances of Siwi genes estimated by 2(-ΔΔCt) method were tested with different HKGs as the reference gene, proving the importance of internal controls in qPCR data analysis. The results not only presented a list of suitable reference genes in four lepidopteran insects, but also proved that the expression stabilities of HKGs were different among evolutionarily close species. There was no single universal reference gene that could be used in all situations. It is indispensable to validate the expression of HKGs before using them as the internal control in qPCR.

  8. Application of propidium monoazide quantitative real-time PCR to quantify the viability of Lactobacillus delbrueckii ssp. bulgaricus.

    Science.gov (United States)

    Shao, Yuyu; Wang, Zhaoxia; Bao, Qiuhua; Zhang, Heping

    2016-12-01

    In this study, a combination of propidium monoazide (PMA) and quantitative real-time PCR (qPCR) was used to develop a method to determine the viability of cells of Lactobacillus delbrueckii ssp. bulgaricus ND02 (L. bulgaricus) that may have entered into a viable but nonculturable state. This can happen due to its susceptibility to cold shock during lyophilization and storage. Propidium monoazide concentration, PMA incubation time, and light exposure time were optimized to fully exploit the PMA-qPCR approach to accurately assess the total number of living L. bulgaricus ND02. Although PMA has little influence on living cells, when concentrations of PMA were higher than 30μg/mL the number of PCR-positive living bacteria decreased from 10 6 to 10 5 cfu/mL in comparison with qPCR enumeration. Mixtures of living and dead cells were used as method verification samples for enumeration by PMA-qPCR, demonstrating that this method was feasible and effective for distinguishing living cells of L. bulgaricus when mixed with a known number of dead cells. We suggest that several conditions need to be studied further before PMA-qPCR methods can be accurately used to distinguish living from dead cells for enumeration under more realistic sampling situations. However, this research provides a rapid way to enumerate living cells of L. bulgaricus and could be used to optimize selection of cryoprotectants in the lyophilization process and develop technologies for high cell density cultivation and optimal freeze-drying processes. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. 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

  10. Real-Time Leaky Lamb Wave Spectrum Measurement and Its Application to NDE of Composites

    Science.gov (United States)

    Lih, Shyh-Shiuh; Bar-Cohen, Yoseph

    1999-01-01

    Numerous analytical and theoretical studies of the behavior of leaky Lamb waves (LLW) in composite materials were documented in the literature. One of the key issues that are constraining the application of this method as a practical tool is the amount of data that needs to be acquired and the slow process that is involved with such experiments. Recently, a methodology that allows quasi real-time acquisition of LLW dispersion data was developed. At each angle of incidence the reflection spectrum is available in real time from the experimental setup and it can be used for rapid detection of the defects. This technique can be used to rapidly acquire the various plate wave modes along various angles of incidence for the characterization of the material elastic properties. The experimental method and data acquisition technique will be described in this paper. Experimental data was used to examine a series of flaws including porosity and delaminations and demonstrated the efficiency of the developed technique.

  11. XpertTrack: Precision Autonomous Measuring Device Developed for Real Time Shipments Tracker

    Directory of Open Access Journals (Sweden)

    Liviu Viman

    2016-03-01

    Full Text Available This paper proposes a software and hardware solution for real time condition monitoring applications. The proposed device, called XpertTrack, exchanges data through the GPRS protocol over a GSM network and monitories temperature and vibrations of critical merchandise during commercial shipments anywhere on the globe. Another feature of this real time tracker is to provide GPS and GSM positioning with a precision of 10 m or less. In order to interpret the condition of the merchandise, the data acquisition, analysis and visualization are done with 0.1 °C accuracy for the temperature sensor, and 10 levels of shock sensitivity for the acceleration sensor. In addition to this, the architecture allows increasing the number and the types of sensors, so that companies can use this flexible solution to monitor a large percentage of their fleet.

  12. XpertTrack: Precision Autonomous Measuring Device Developed for Real Time Shipments Tracker.

    Science.gov (United States)

    Viman, Liviu; Daraban, Mihai; Fizesan, Raul; Iuonas, Mircea

    2016-03-10

    This paper proposes a software and hardware solution for real time condition monitoring applications. The proposed device, called XpertTrack, exchanges data through the GPRS protocol over a GSM network and monitories temperature and vibrations of critical merchandise during commercial shipments anywhere on the globe. Another feature of this real time tracker is to provide GPS and GSM positioning with a precision of 10 m or less. In order to interpret the condition of the merchandise, the data acquisition, analysis and visualization are done with 0.1 °C accuracy for the temperature sensor, and 10 levels of shock sensitivity for the acceleration sensor. In addition to this, the architecture allows increasing the number and the types of sensors, so that companies can use this flexible solution to monitor a large percentage of their fleet.

  13. XpertTrack: Precision Autonomous Measuring Device Developed for Real Time Shipments Tracker

    Science.gov (United States)

    Viman, Liviu; Daraban, Mihai; Fizesan, Raul; Iuonas, Mircea

    2016-01-01

    This paper proposes a software and hardware solution for real time condition monitoring applications. The proposed device, called XpertTrack, exchanges data through the GPRS protocol over a GSM network and monitories temperature and vibrations of critical merchandise during commercial shipments anywhere on the globe. Another feature of this real time tracker is to provide GPS and GSM positioning with a precision of 10 m or less. In order to interpret the condition of the merchandise, the data acquisition, analysis and visualization are done with 0.1 °C accuracy for the temperature sensor, and 10 levels of shock sensitivity for the acceleration sensor. In addition to this, the architecture allows increasing the number and the types of sensors, so that companies can use this flexible solution to monitor a large percentage of their fleet. PMID:26978360

  14. Simulation of a nuclear measurement system around a multi-task mode real-time monitor

    International Nuclear Information System (INIS)

    De Grandi, G.; Ouiguini, R.

    1983-01-01

    When debugging and testing material and software for the automation of systems, the non-availability of this last one states important logistic problems. A simulator of the system to be automatized, conceived around a multi-task mode real-time monitor, allowing the debugging of the software of automation without the physical presence of the system to be automatized, is proposed in the present report

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. Validation of reference genes for quantitative real-time PCR in Périgord black truffle (Tuber melanosporum) developmental stages.

    Science.gov (United States)

    Zarivi, Osvaldo; Cesare, Patrizia; Ragnelli, Anna Maria; Aimola, Pierpaolo; Leonardi, Marco; Bonfigli, Antonella; Colafarina, Sabrina; Poma, Anna Maria; Miranda, Michele; Pacioni, Giovanni

    2015-08-01

    The symbiotic fungus Tuber melanosporum Vittad. (Périgord black truffle) belongs to the Ascomycota and forms mutualistic symbiosis with tree and shrub roots. This truffle has a high value in a global market and is cultivated in many countries of both hemispheres. The publication of the T. melanosporum genome has given researchers unique opportunities to learn more about the biology of the fungus. Real-time quantitative PCR (qRT-PCR) is a definitive technique for quantitating differences in transcriptional gene expression levels between samples. To facilitate gene expression studies and obtain more accurate qRT-PCR data, normalization relative to stable housekeeping genes is required. These housekeeping genes must show stable expression under given experimental conditions for the qRT-PCR results to be accurate. Unfortunately, there are no studies on the stability of housekeeping genes used in T. melanosporum development. In this study, we present a morphological and microscopical classification of the developmental stages of T. melanosporum fruit body, and investigate the expression levels of 12 candidate reference genes (18S rRNA; 5.8S rRNA; Elongation factor 1-alpha; Elongation factor 1-beta; α-tubulin; 60S ribosomal protein L29; β-tubulin; 40S ribosomal protein S1; 40S ribosomal protein S3; Glucose-6-phosphate dehydrogenase; β-actin; Ubiquitin-conjugating enzyme). To evaluate the suitability of these genes as endogenous controls, five software-based approaches and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes. We demonstrate here that the 18S rRNA gene shows the most stable expression during T. melanosporum development and that a set of three genes, 18S rRNA, Elongation factor 1-alpha and 40S ribosomal protein S3, is the most suitable to normalize qRT-PCR data from all the analyzed developmental stages; conversely, 18S rRNA, Glucose-6-phosphate dehydrogenase and Elongation factor 1-alpha are the most suitable

  20. Evaluation and validation of candidate endogenous control genes for real-time quantitative PCR studies of breast cancer

    Directory of Open Access Journals (Sweden)

    Miller Nicola

    2007-11-01

    Full Text Available Abstract Background Real-time quantitative PCR (RQ-PCR forms the basis of many breast cancer biomarker studies and novel prognostic assays, paving the way towards personalised cancer treatments. Normalisation of relative RQ-PCR data is required to control for non-biological variation introduced during sample preparation. Endogenous control (EC genes, used in this context, should ideally be expressed constitutively and uniformly across treatments in all test samples. Despite widespread recognition that the accuracy of the normalised data is largely dependent on the reliability of the EC, there are no reports of the systematic validation of genes commonly used for this purpose in the analysis of gene expression by RQ-PCR in primary breast cancer tissues. The aim of this study was to identify the most suitable endogenous control genes for RQ-PCR analysis of primary breast tissue from a panel of eleven candidates in current use. Oestrogen receptor alpha (ESR1 was used a target gene to compare the effect of choice of EC on the estimate of gene quantity. Results The expression and validity of candidate ECs (GAPDH, TFRC, ABL, PPIA, HPRT1, RPLP0, B2M, GUSB, MRPL19, PUM1 and PSMC4 was determined in 6 benign and 21 malignant primary breast cancer tissues. Gene expression data was analysed using two different statistical models. MRPL19 and PPIA were identified as the most stable and reliable EC genes, while GUSB, RPLP0 and ABL were least stable. There was a highly significant difference in variance between ECs. ESR1 expression was appreciably higher in malignant compared to benign tissues and there was a significant effect of EC on the magnitude of the error associated with the relative quantity of ESR1. Conclusion We have validated two endogenous control genes, MRPL19 and PPIA, for RQ-PCR analysis of gene expression in primary breast tissue. Of the genes in current use in this field, the above combination offers increased accuracy and resolution in the

  1. Isothermal microcalorimetry - A quantitative method to monitor Trypanosoma congolense growth and growth inhibition by trypanocidal drugs in real time.

    Science.gov (United States)

    Gysin, M; Braissant, O; Gillingwater, K; Brun, R; Mäser, P; Wenzler, T

    2018-03-16

    Trypanosoma congolense is a protozoan parasite that is transmitted by tsetse flies, causing African Animal Trypanosomiasis, also known as Nagana, in sub-Saharan Africa. Nagana is a fatal disease of livestock that causes severe economic losses. Two drugs are available, diminazene and isometamidium, yet successful treatment is jeopardized by drug resistant T. congolense. Isothermal microcalorimetry is a highly sensitive tool that can be used to study growth of the extracellular T. congolense parasites or to study parasite growth inhibition after the addition of antitrypanosomal drugs. Time of drug action and time to kill can be quantified in a simple way by real time heat flow measurements. We established a robust protocol for the microcalorimetric studies of T. congolense and developed mathematical computations in R to calculate different parameters related to growth and the kinetics of drug action. We demonstrate the feasibility and benefit of the method exemplary with the two standard drugs, diminazene aceturate and isometamidium chloride. The method and the mathematical approach can be translated to study other pathogenic or non-pathogenic cells if they are metabolically active and grow under axenic conditions. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. 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.

  3. 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.

  4. 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.

  5. Evaluation of some software measuring displacements using GPS in real-time

    Science.gov (United States)

    Langbein, John

    2006-01-01

    For the past decade, the USGS has been monitoring deformation at various locations in the western United States using continuous GPS. The main focus of these measurements are estimates of displacement averaged over one day. Essentially, these consist of recording at 30 seconds intervals the carrier-frequency phase-data (equivalent to travel-time) between a GPS receiver and the GPS satellite network. In turn, these observations, which are converted to pseudo—ranges, are processed using one of the “research grade” programs (GIPSY, Zumberge et al., or GAMIT, wwwgpsg.mit.edu/~simon/gtgk) to estimate the position of the GPS receiver averaged over 24 hours. However, it is possible and desirable to estimate the position of the receiver (actually the antenna) more frequently and to do this within a few seconds of the time actual measurement (known as real-time). A recent example, the 2004 Magnitude 6, Parkfield, California earthquake, demonstrated that having GPS estimates of position more frequently than simply a daily average is required if one requires discrimination between co-seismic and post-seismic deformation (Langbein et al., 2006). The high-rate estimates of position obtained at Parkfield show that post-seismic deformation started less than one-hour after the mainshock and that this deformation was roughly the same magnitude as the co-seismic deformation. The high-rate solutions for Parkfield were done by others including Yehuda Bock at UCSD and Kristine Larson at U. of Colorado, but not the USGS. The Parkfield experience points out the need for an in-house capability by the USGS to be able to accurately measure co-seismic displacements and other rapid, deformation signals using GPS. This applies to both the Earthquake and Volcano Hazard programs. Although at many locations where we monitor deformation, we have strainmeters and tiltmeters in addition to GPS which, in principle, are far more sensitive to rapid deformation over periods of less than a day

  6. A real time measurement of junction temperature variation in high power IGBT modules for wind power converter application

    DEFF Research Database (Denmark)

    Ghimire, Pramod; Pedersen, Kristian Bonderup; de Vega, Angel Ruiz

    2014-01-01

    This paper presents a real time measurement of on-state forward voltage and estimating the junction temperature for a high power IGBT module during a power converter operation. The power converter is realized as it can be used for a wind turbine system. The peak of the junction temperature is dec...

  7. In-situ real time measurements of thermal comfort and comparison with the adaptive comfort theory in Dutch residential dwellings

    NARCIS (Netherlands)

    Ioannou, A.; Itard, L.C.M.; Agarwal, Tushar

    2018-01-01

    Indoor thermal comfort is generally assessed using the PMV or the adaptive model. This research presents the results obtained by in-situ real time measurements of thermal comfort and thermal comfort perception in 17 residential dwellings in the Netherlands. The study demonstrates the new

  8. REAL-TIME MEASUREMENT OF AIRWAY RESPONSES TO SULOFUR DIOXIDE (SO2) IN AN INTACT, AWAKE GUINEA PIG MODEL

    Science.gov (United States)

    Real-time measurment of airway responses to Sulfur Dioxide (SO2) in an intact, awake guinea pig model. J Stanek1,2, Q Krantz2, J Nolan2, D Winsett2, W Watkinson2, and D Costa2. 1College of Veterinary Medicine, NCSU, Raleigh, NC, USA; 2Pulmonary Toxicology Branch, ETD, NHEERL, US...

  9. 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

  10. 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

  11. 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

  12. 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...

  13. 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...

  14. LOS Throughput Measurements in Real-Time with a 128-Antenna Massive MIMO Testbed

    OpenAIRE

    Harris, Paul; Zhang, Siming; Beach, Mark; Mellios, Evangelos; Nix, Andrew; Armour, Simon; Doufexi, Angela; Nieman, Karl; Kundargi, Nikhil

    2017-01-01

    This paper presents initial results for a novel 128-antenna massive Multiple-Input, Multiple- Output (MIMO) testbed developed through Bristol Is Open in collaboration with National Instruments and Lund University. We believe that the results presented here validate the adoption of massive MIMO as a key enabling technology for 5G and pave the way for further pragmatic research by the massive MIMO community. The testbed operates in real-time with a Long-Term Evolution (LTE)-like PHY in Time Div...

  15. Assessment of offsite, real-time dose measurement systems for emergency situations

    International Nuclear Information System (INIS)

    Maeck, W.J.; Hoffman, L.G.; Staples, B.A.; Keller, J.H.

    1982-04-01

    An evaluation is made of the effectiveness of fixed, real-time monitoring systems around nuclear power stations in determining the magnitude of unmonitored releases. The effects of meteorological conditions on the accuracy with which the magnitude of unmonitored releases is determined and the uncertainties inherent in defining these meteorological conditions are discussed. The number and placement of fixed field detectors in a system is discussed, and the data processing equipment required to convert field detector output data into release rate information is described. Cost data relative to the purchase and installation of specific systems are given, as well as the characteristics and information return for a system purchased at an arbitrary cost

  16. Mobile laboratory for near real-time measurements of very low-level radioactivity

    International Nuclear Information System (INIS)

    Sigg, R.A.

    1984-01-01

    The Tracking Radioactive Atmospheric Contaminants (TRAC) System is a mobile laboratory, developed by Savannah River Laboratory (SRL) to improve emergency response and environmental research capabilities at the Savannah River Plant (SRP). In the event of an atmospheric release, the TRAC laboratory can confirm the location and radionuclide composition of the downwind cloud by analyzing samples in near real-time in the field. Specialized monitoring systems were developed to analyze most radionuclides produced in SRP's diverse operations. Sensitivities are radionuclide dependent and can be below maximum permissible concentration (MPC) values by factors as large as one hundred thousand. 6 references, 6 figures

  17. Human perception of the measurement of a network attack taxonomy in near real-time

    CSIR Research Space (South Africa)

    van Heerden, R

    2014-07-01

    Full Text Available apparent long after an attack and even then it is sometimes only speculation. For example, the Aggressor can not be determined in a near real-time environment and for some attacks the real power behind the attack is never determined or proven. 2.1 Actor...- tigating the accuracy of various assessment methods. We classify each class as either defined, high, low or not quantifiable. For example, it may not be possi- ble to determine the instigator of an attack (Aggressor), but only that the attack has been...

  18. Comparison of lung cancer cell lines representing four histopathological subtypes with gene expression profiling using quantitative real-time PCR

    Directory of Open Access Journals (Sweden)

    Kawaguchi Makoto

    2010-01-01

    Full Text Available Abstract Background Lung cancers are the most common type of human malignancy and are intractable. Lung cancers are generally classified into four histopathological subtypes: adenocarcinoma (AD, squamous cell carcinoma (SQ, large cell carcinoma (LC, and small cell carcinoma (SC. Molecular biological characterization of these subtypes has been performed mainly using DNA microarrays. In this study, we compared the gene expression profiles of these four subtypes using twelve human lung cancer cell lines and the more reliable quantitative real-time PCR (qPCR. Results We selected 100 genes from public DNA microarray data and examined them by DNA microarray analysis in eight test cell lines (A549, ABC-1, EBC-1, LK-2, LU65, LU99, STC 1, RERF-LC-MA and a normal control lung cell line (MRC-9. From this, we extracted 19 candidate genes. We quantified the expression of the 19 genes and a housekeeping gene, GAPDH, with qPCR, using the same eight cell lines plus four additional validation lung cancer cell lines (RERF-LC-MS, LC-1/sq, 86-2, and MS-1-L. Finally, we characterized the four subtypes of lung cancer cell lines using principal component analysis (PCA of gene expression profiling for 12 of the 19 genes (AMY2A, CDH1, FOXG1, IGSF3, ISL1, MALL, PLAU, RAB25, S100P, SLCO4A1, STMN1, and TGM2. The combined PCA and gene pathway analyses suggested that these genes were related to cell adhesion, growth, and invasion. S100P in AD cells and CDH1 in AD and SQ cells were identified as candidate markers of these lung cancer subtypes based on their upregulation and the results of PCA analysis. Immunohistochemistry for S100P and RAB25 was closely correlated to gene expression. Conclusions These results show that the four subtypes, represented by 12 lung cancer cell lines, were well characterized using qPCR and PCA for the 12 genes examined. Certain genes, in particular S100P and CDH1, may be especially important for distinguishing the different subtypes. Our results

  19. O-5S quantitative real-time PCR: a new diagnostic tool for laboratory confirmation of human onchocerciasis.

    Science.gov (United States)

    Mekonnen, Solomon A; Beissner, Marcus; Saar, Malkin; Ali, Solomon; Zeynudin, Ahmed; Tesfaye, Kassahun; Adbaru, Mulatu G; Battke, Florian; Poppert, Sven; Hoelscher, Michael; Löscher, Thomas; Bretzel, Gisela; Herbinger, Karl-Heinz

    2017-10-02

    Onchocerciasis is a parasitic disease caused by the filarial nematode Onchocerca volvulus. In endemic areas, the diagnosis is commonly confirmed by microscopic examination of skin snip samples, though this technique is considered to have low sensitivity. The available melting-curve based quantitative real-time PCR (qPCR) using degenerated primers targeting the O-150 repeat of O. volvulus was considered insufficient for confirming the individual diagnosis, especially in elimination studies. This study aimed to improve detection of O. volvulus DNA in clinical samples through the development of a highly sensitive qPCR assay. A novel hydrolysis probe based qPCR assay was designed targeting the specific sequence of the O. volvulus O-5S rRNA gene. A total of 200 clinically suspected onchocerciasis cases were included from Goma district in South-west Ethiopia, from October 2012 through May 2013. Skin snip samples were collected and subjected to microscopy, O-150 qPCR, and the novel O-5S qPCR. Among the 200 individuals, 133 patients tested positive (positivity rate of 66.5%) and 67 negative by O-5S qPCR, 74 tested positive by microscopy (37.0%) and 78 tested positive by O-150 qPCR (39.0%). Among the 133 O-5S qPCR positive individuals, microscopy and O-150 qPCR detected 55.6 and 59.4% patients, respectively, implying a higher sensitivity of O-5S qPCR than microscopy and O-150 qPCR. None of the 67 individuals who tested negative by O-5S qPCR tested positive by microscopy or O-150 qPCR, implying 100% specificity of the newly designed O-5S qPCR assay. The novel O-5S qPCR assay is more sensitive than both microscopic examination and the existing O-150 qPCR for the detection of O. volvulus from skin snip samples. The newly designed assay is an important step towards appropriate individual diagnosis and control of onchocerciasis.

  20. Evaluation of reference genes for quantitative real-time PCR in oil palm elite planting materials propagated by tissue culture.

    Directory of Open Access Journals (Sweden)

    Pek-Lan Chan

    Full Text Available BACKGROUND: The somatic embryogenesis tissue culture process has been utilized to propagate high yielding oil palm. Due to the low callogenesis and embryogenesis rates, molecular studies were initiated to identify genes regulating the process, and their expression levels are usually quantified using reverse transcription quantitative real-time PCR (RT-qPCR. With the recent release of oil palm genome sequences, it is crucial to establish a proper strategy for gene analysis using RT-qPCR. Selection of the most suitable reference genes should be performed for accurate quantification of gene expression levels. RESULTS: In this study, eight candidate reference genes selected from cDNA microarray study and literature review were evaluated comprehensively across 26 tissue culture samples using RT-qPCR. These samples were collected from two tissue culture lines and media treatments, which consisted of leaf explants cultures, callus and embryoids from consecutive developmental stages. Three statistical algorithms (geNorm, NormFinder and BestKeeper confirmed that the expression stability of novel reference genes (pOP-EA01332, PD00380 and PD00569 outperformed classical housekeeping genes (GAPDH, NAD5, TUBULIN, UBIQUITIN and ACTIN. PD00380 and PD00569 were identified as the most stably expressed genes in total samples, MA2 and MA8 tissue culture lines. Their applicability to validate the expression profiles of a putative ethylene-responsive transcription factor 3-like gene demonstrated the importance of using the geometric mean of two genes for normalization. CONCLUSIONS: Systematic selection of the most stably expressed reference genes for RT-qPCR was established in oil palm tissue culture samples. PD00380 and PD00569 were selected for accurate and reliable normalization of gene expression data from RT-qPCR. These data will be valuable to the research associated with the tissue culture process. Also, the method described here will facilitate the selection

  1. Evaluation of reference genes for real-time quantitative PCR studies in Candida glabrata following azole treatment

    Directory of Open Access Journals (Sweden)

    Li Qingdi

    2012-06-01

    Full Text Available Abstract Background The selection of stable and suitable reference genes for real-time quantitative PCR (RT-qPCR is a crucial prerequisite for reliable gene expression analysis under different experimental conditions. The present study aimed to identify reference genes as internal controls for gene expression studies by RT-qPCR in azole-stimulated Candida glabrata. Results The expression stability of 16 reference genes under fluconazole stress was evaluated using fold change and standard deviation computations with the hkgFinder tool. Our data revealed that the mRNA expression levels of three ribosomal RNAs (RDN5.8, RDN18, and RDN25 remained stable in response to fluconazole, while PGK1, UBC7, and UBC13 mRNAs showed only approximately 2.9-, 3.0-, and 2.5-fold induction by azole, respectively. By contrast, mRNA levels of the other 10 reference genes (ACT1, EF1α, GAPDH, PPIA, RPL2A, RPL10, RPL13A, SDHA, TUB1, and UBC4 were dramatically increased in C. glabrata following antifungal treatment, exhibiting changes ranging from 4.5- to 32.7-fold. We also assessed the expression stability of these reference genes using the 2-ΔΔCT method and three other software packages. The stability rankings of the reference genes by geNorm and the 2-ΔΔCT method were identical to those by hkgFinder, whereas the stability rankings by BestKeeper and NormFinder were notably different. We then validated the suitability of six candidate reference genes (ACT1, PGK1, RDN5.8, RDN18, UBC7, and UBC13 as internal controls for ten target genes in this system using the comparative CT method. Our validation experiments passed for all six reference genes analyzed except RDN18, where the amplification efficiency of RDN18 was different from that of the ten target genes. Finally, we demonstrated that the relative quantification of target gene expression varied according to the endogenous control used, highlighting the importance of the choice of internal controls in such

  2. Evaluation of Reference Genes for Quantitative Real-Time PCR in Oil Palm Elite Planting Materials Propagated by Tissue Culture

    Science.gov (United States)

    Chan, Pek-Lan; Rose, Ray J.; Abdul Murad, Abdul Munir; Zainal, Zamri; Leslie Low, Eng-Ti; Ooi, Leslie Cheng-Li; Ooi, Siew-Eng; Yahya, Suzaini; Singh, Rajinder

    2014-01-01

    Background The somatic embryogenesis tissue culture process has been utilized to propagate high yielding oil palm. Due to the low callogenesis and embryogenesis rates, molecular studies were initiated to identify genes regulating the process, and their expression levels are usually quantified using reverse transcription quantitative real-time PCR (RT-qPCR). With the recent release of oil palm genome sequences, it is crucial to establish a proper strategy for gene analysis using RT-qPCR. Selection of the most suitable reference genes should be performed for accurate quantification of gene expression levels. Results In this study, eight candidate reference genes selected from cDNA microarray study and literature review were evaluated comprehensively across 26 tissue culture samples using RT-qPCR. These samples were collected from two tissue culture lines and media treatments, which consisted of leaf explants cultures, callus and embryoids from consecutive developmental stages. Three statistical algorithms (geNorm, NormFinder and BestKeeper) confirmed that the expression stability of novel reference genes (pOP-EA01332, PD00380 and PD00569) outperformed classical housekeeping genes (GAPDH, NAD5, TUBULIN, UBIQUITIN and ACTIN). PD00380 and PD00569 were identified as the most stably expressed genes in total samples, MA2 and MA8 tissue culture lines. Their applicability to validate the expression profiles of a putative ethylene-responsive transcription factor 3-like gene demonstrated the importance of using the geometric mean of two genes for normalization. Conclusions Systematic selection of the most stably expressed reference genes for RT-qPCR was established in oil palm tissue culture samples. PD00380 and PD00569 were selected for accurate and reliable normalization of gene expression data from RT-qPCR. These data will be valuable to the research associated with the tissue culture process. Also, the method described here will facilitate the selection of appropriate

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

    Directory of Open Access Journals (Sweden)

    Qiusheng Kong

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

  4. DEVELOPMENT OF AN ON-LINE, REAL-TIME ALPHA RADIATION MEASURING INSTRUMENT FOR LIQUID STREAMS

    International Nuclear Information System (INIS)

    Unknown

    1999-01-01

    Thermo Power Corporation has proven the technical viability of an on-line, real-time alpha radionuclide instrument for aqueous sample analysis through laboratory and initial field tests of the instrument. The instrument has been shown to be isotonically sensitive to extremely low (ten parts per trillion, or femto Curies per liter) levels of a broad range of radioisotopes. Performance enhancement and other scaling data obtained during the course of this investigation have shown that on-line, real-time operation is possible, with a sub 30-minute response time analyzing 20 ppb (30 pCi/1) natural uranium. Now that these initial field tests in Oak Ridge, Tennessee have been successfully completed, Thermo Power plans to conduct comprehensive field tests of the instrument. The purpose of these endurance tests will be to determine the endurance characteristics of the Thermo Alpha Monitor for Water when it is used by non-Thermo Power personnel in a series of one or more extended field tests. Such endurance testing is the vital next step towards the commercialization of the Alpha Monitor. Subsequently, it will be possible to provide the DOE with an instrument that has the capability of obtaining rapid feedback about the concentrations of alpha-emitting isotope contamination in effluent water streams (Subsurface Contaminants Focus Area). It will also be useful for process control of remediation and D and D operations such as monitoring scrubber/rinse water radioactivity levels (Mixed Waste, Plutonium and D and D Focus Areas)

  5. An FPGA Architecture for Extracting Real-Time Zernike Coefficients from Measured Phase Gradients

    Science.gov (United States)

    Moser, Steven; Lee, Peter; Podoleanu, Adrian

    2015-04-01

    Zernike modes are commonly used in adaptive optics systems to represent optical wavefronts. However, real-time calculation of Zernike modes is time consuming due to two factors: the large factorial components in the radial polynomials used to define them and the large inverse matrix calculation needed for the linear fit. This paper presents an efficient parallel method for calculating Zernike coefficients from phase gradients produced by a Shack-Hartman sensor and its real-time implementation using an FPGA by pre-calculation and storage of subsections of the large inverse matrix. The architecture exploits symmetries within the Zernike modes to achieve a significant reduction in memory requirements and a speed-up of 2.9 when compared to published results utilising a 2D-FFT method for a grid size of 8×8. Analysis of processor element internal word length requirements show that 24-bit precision in precalculated values of the Zernike mode partial derivatives ensures less than 0.5% error per Zernike coefficient and an overall error of RAM usage is <16% for Shack-Hartmann grid sizes up to 32×32.

  6. A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR

    NARCIS (Netherlands)

    J.P.P. Meijerink (Jules); C. Mandigers; L. van de Locht; E. Tonnissen; F. Goodsaid; J. Raemaekers (John)

    2001-01-01

    textabstractQuantification of residual disease by real-time polymerase chain reaction (PCR) will become a pivotal tool in the development of patient-directed therapy. In recent years, various protocols to quantify minimal residual disease in leukemia or lymphoma patients have been

  7. A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR.

    NARCIS (Netherlands)

    Meijerink, J.P.P.; Mandigers, C.M.P.W.; Locht, A.T.F. van de; Tonnissen, E.L.R.T.M.; Goodsaid, F.; Raemaekers, J.M.M.

    2001-01-01

    Quantification of residual disease by real-time polymerase chain reaction (PCR) will become a pivotal tool in the development of patient-directed therapy. In recent years, various protocols to quantify minimal residual disease in leukemia or lymphoma patients have been developed. These assays assume

  8. Partial validation of a TaqMan real-time quantitative PCR for the detection of ranaviruses

    DEFF Research Database (Denmark)

    Stilwell, Natalie K.; Whittington, Richard J.; Hick, Paul M.

    2018-01-01

    Ranaviruses are globally emerging pathogens negatively impacting wild and cultured fish, amphibians, and reptiles. Although conventional and diagnostic real-time PCR (qPCR) assays have been developed to detect ranaviruses, these assays often have not been tested against the known diversity of ran...

  9. User satisfaction with a real-time automated feedback system for general practitioners: a quantitative and qualitative study

    NARCIS (Netherlands)

    Bindels, Rianne; Hasman, Arie; Derickx, Mieke; van Wersch, Jan W. J.; Winkens, Ron A. G.

    2003-01-01

    OBJECTIVE: The GRIF automated feedback system produces real-time comments on the appropriateness of diagnostic tests ordered by general practitioners (GPs) based on recommendations from accepted national and regional practice guidelines. We investigated the experiences of GPs with this system and,

  10. Quantitative analysis of the improvement in high zoom maritime tracking due to real-time image enhancement

    CSIR Research Space (South Africa)

    Bachoo, AK

    2011-04-01

    Full Text Available This work aims to evaluate the improvement in the performance of tracking small maritime targets due to real-time enhancement of the video streams from high zoom cameras on pan-tilt pedestal. Due to atmospheric conditions these images can frequently...

  11. Two quantitative real-time PCR assays for the detection of penaeid shrimp and blue crab, crustacean shellfish allergens.

    Science.gov (United States)

    Eischeid, Anne C; Kim, Bang-hyun; Kasko, Sasha M

    2013-06-19

    Food allergen detection methods must be able to specifically detect minute quantities of an allergenic food in a complex food matrix. One technique that can be used is real-time PCR. For the work described here, real-time PCR assays were developed to detect penaeid shrimp and blue crab, crustacean shellfish allergens. The method was tested using shrimp meat and crab meat spiked into several types of foods, including canned soups, deli foods, meat, seafood, and prepared seafood products. Foods were spiked with either shrimp or crab at levels ranging from 0.1 to 10⁶ parts per million (ppm) and analyzed either raw or cooked by a variety of methods. Real-time PCR data were used to generate linear standard curves, and assays were evaluated with respect to linear range and reaction efficiency. Results indicate that both assays performed well in a variety of food types. High reaction efficiencies were achieved across a linear range of 6-8 orders of magnitude. Limits of detection were generally between 0.1 and 1 ppm. Cooking methods used to simulate thermal processing of foods had little effect on assay performance. This work demonstrates that real-time PCR can be a valuable tool in the detection of crustacean shellfish.

  12. 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.

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

    International Nuclear Information System (INIS)

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

    2008-01-01

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

  14. 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

  15. 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

  16. 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)

  17. Measurement and analysis of workload effects on fault latency in real-time systems

    Science.gov (United States)

    Woodbury, Michael H.; Shin, Kang G.

    1990-01-01

    The authors demonstrate the need to address fault latency in highly reliable real-time control computer systems. It is noted that the effectiveness of all known recovery mechanisms is greatly reduced in the presence of multiple latent faults. The presence of multiple latent faults increases the possibility of multiple errors, which could result in coverage failure. The authors present experimental evidence indicating that the duration of fault latency is dependent on workload. A synthetic workload generator is used to vary the workload, and a hardware fault injector is applied to inject transient faults of varying durations. This method makes it possible to derive the distribution of fault latency duration. Experimental results obtained from the fault-tolerant multiprocessor at the NASA Airlab are presented and discussed.

  18. Real-time detection of antibiotic activity by measuring nanometer-scale bacterial deformation

    Science.gov (United States)

    Iriya, Rafael; Syal, Karan; Jing, Wenwen; Mo, Manni; Yu, Hui; Haydel, Shelley E.; Wang, Shaopeng; Tao, Nongjian

    2017-12-01

    Diagnosing antibiotic-resistant bacteria currently requires sensitive detection of phenotypic changes associated with antibiotic action on bacteria. Here, we present an optical imaging-based approach to quantify bacterial membrane deformation as a phenotypic feature in real-time with a nanometer scale (˜9 nm) detection limit. Using this approach, we found two types of antibiotic-induced membrane deformations in different bacterial strains: polymyxin B induced relatively uniform spatial deformation of Escherichia coli O157:H7 cells leading to change in cellular volume and ampicillin-induced localized spatial deformation leading to the formation of bulges or protrusions on uropathogenic E. coli CFT073 cells. We anticipate that the approach will contribute to understanding of antibiotic phenotypic effects on bacteria with a potential for applications in rapid antibiotic susceptibility testing.

  19. 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...

  20. 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)

  1. 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.

  2. 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.

  3. High-efficient method for spectrometric data real time processing with increased resolution of a measuring channel

    International Nuclear Information System (INIS)

    Ashkinaze, S.I.; Voronov, V.A.; Nechaev, Yu.I.

    1988-01-01

    Solution of reduction problem as a mean to increase spectrometric tract resolution when it is realized using the digit-by-digit modified method and special strategy, significantly reducing the time of processing, is considered. The results presented confirm that the complex measurement tract plus microcomputer is equivalent to the use of the tract with a higher resolution, and the use of the digit-by-digit modified method permits to process spectrometric information in real time scale

  4. An amperometric enzyme biosensor for real-time measurements of cellobiohydrolase activity on insoluble cellulose

    DEFF Research Database (Denmark)

    Cruys-Bagger, Nicolaj; Guilin, Ren; Tatsumi, Hirosuke

    2012-01-01

    An amperometric enzyme biosensor for continuous detection of cellobiose has been implemented as an enzyme assay for cellulases. We show that the initial kinetics for cellobiohydrolase I, Cel7A from Trichoderma reesei, acting on different types of cellulose substrates, semi-crystalline and amorphous......, can be monitored directly and in real-time by an enzyme-modified electrode based on cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium (Pc). PcCDH was cross-linked and immobilized on the surface of a carbon paste electrode which contained a mediator, benzoquinone. An oxidation current...... of the reduced mediator, hydroquinone, produced by the CDH-catalyzed reaction with cellobiose, was recorded under constant-potential amperometry at +0.5 V (vs. Ag/AgCl). The CDH-biosensors showed high sensitivity (87.7 µA mM−1 cm−2), low detection limit (25 nM), and fast response time (t95% ∼ 3 s...

  5. Measuring Gait Quality in Parkinson’s Disease through Real-Time Gait Phase Recognition

    Directory of Open Access Journals (Sweden)

    Ilaria Mileti

    2018-03-01

    Full Text Available Monitoring gait quality in daily activities through wearable sensors has the potential to improve medical assessment in Parkinson’s Disease (PD. In this study, four gait partitioning methods, two based on thresholds and two based on a machine learning approach, considering the four-phase model, were compared. The methods were tested on 26 PD patients, both in OFF and ON levodopa conditions, and 11 healthy subjects, during walking tasks. All subjects were equipped with inertial sensors placed on feet. Force resistive sensors were used to assess reference time sequence of gait phases. Goodness Index (G was evaluated to assess accuracy in gait phases estimation. A novel synthetic index called Gait Phase Quality Index (GPQI was proposed for gait quality assessment. Results revealed optimum performance (G < 0.25 for three tested methods and good performance (0.25 < G < 0.70 for one threshold method. The GPQI resulted significantly higher in PD patients than in healthy subjects, showing a moderate correlation with clinical scales score. Furthermore, in patients with severe gait impairment, GPQI was found higher in OFF than in ON state. Our results unveil the possibility of monitoring gait quality in PD through real-time gait partitioning based on wearable sensors.

  6. The Real-Time Dose Measurement Scintillating Fiber Array for Brachytherapy Procedures

    Science.gov (United States)

    Tynes, Lawrence

    2007-03-01

    Brachytherapy is a treatment modality that uses tiny radioactive sources (few mm in length) by delivering enough doses to kill cancer tumors or plaque build-up. The type of sources used in hospitals include both gamma and beta emitters. Presently, the technique suffers from not having a single detector with the capability of providing accurate dose distribution information within sub-mm accuracy. The current standard is based primarily on well chambers and film dosimetry. The Center for Advanced Medical Instrumentation (CAMI) at Hampton University is developing a Scintillating Fiber Based Beta Detector prototype in collaboration with the National Institute for Standards and Technology (NIST) to address this problem. The device is composed of an array of 1x1 mm^2 scintillating fibers optically coupled to photo-multiplier tubes for photon-to-current conversion. A CAMAC LabView based data acquisition system is used for real time data collection and histogramming, data analysis. A set of data were collected at the nearby Bon Secours DePaul Medical Center using a GammaMed 12i HDR after-loader housing a 6.62 mCi Ir-192 source. Preliminary comparison between our device and film dosimetry will be discussed.

  7. Measuring Chemotherapy Appointment Duration and Variation Using Real-Time Location Systems.

    Science.gov (United States)

    Barysauskas, Constance M; Hudgins, Gina; Gill, Katie Kupferberg; Camuso, Kristen M; Bagley, Janet; Rozanski, Sheila; Kadish, Sarah

    Clinical schedules drive resource utilization, cost, and patient wait time. Accurate appointment duration allocation ensures appropriate staffing ratios to daily caseloads and maximizes scarce resources. Dana-Farber Cancer Institute (DFCI) infusion appointment duration is adjusted by regimen using a consensus method of experts including pharmacists, nurses, and administrators. Using real-time location system (RTLS), we examined the accuracy of observed appointment duration compared with the scheduled duration. Appointment duration was calculated using RTLS at DFCI between August 1, 2013, and September 30, 2013. Duration was defined as the total time a patient occupied an infusion chair. The top 10 administered infusion regimens were investigated (n = 805). Median observed appointment durations were statistically different than the scheduled durations. Appointment durations were shorter than scheduled 98% (C), 95% (I), and 75% (F) of the time and longer than scheduled 77% (A) and 76% (G) of the time. Fifty-six percent of the longer than scheduled (A) appointments were at least 30 minute longer. RTLS provides reliable and unbiased data to improve schedule accuracy. Replacing consensus with system-based data may improve clinic flow, relieve staff stress, and increase patient satisfaction. Further investigation is warranted to elucidate factors that impact variation in appointment duration.

  8. 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.

  9. 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.

  10. A review on real time physical measurement techniques and their attempt to predict wear-out status of IGBT

    DEFF Research Database (Denmark)

    Ghimire, Pramod; Beczkowski, Szymon; Munk-Nielsen, Stig

    2013-01-01

    Insulated Gate Bipolar Transistors (IGBTs) are key component in power converters. Reliability of power converters depend on wear-out process of power modules. A physical parameter such as the on-state collector-emitter voltage (Vce) shows the status of degradation of the IGBT after a certain cycles...... of difficulties in the measurement, the offline and online Vce measurement topologies are implemented to study the reliability of the power converters. This paper presents a review in wear-out prediction methods of IGBT power modules and freewheeling diodes based on the real time Vce measurement. The measurement...

  11. 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.

  12. 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...

  13. 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...

  14. Benchmarking flood models from space in near real-time: accommodating SRTM height measurement errors with low resolution flood imagery

    Science.gov (United States)

    Schumann, G.; di Baldassarre, G.; Alsdorf, D.; Bates, P. D.

    2009-04-01

    In February 2000, the Shuttle Radar Topography Mission (SRTM) measured the elevation of most of the Earth's surface with spatially continuous sampling and an absolute vertical accuracy greater than 9 m. The vertical error has been shown to change with topographic complexity, being less important over flat terrain. This allows water surface slopes to be measured and associated discharge volumes to be estimated for open channels in large basins, such as the Amazon. Building on these capabilities, this paper demonstrates that near real-time coarse resolution radar imagery of a recent flood event on a 98 km reach of the River Po (Northern Italy) combined with SRTM terrain height data leads to a water slope remarkably similar to that derived by combining the radar image with highly accurate airborne laser altimetry. Moreover, it is shown that this space-borne flood wave approximation compares well to a hydraulic model and thus allows the performance of the latter, calibrated on a previous event, to be assessed when applied to an event of different magnitude in near real-time. These results are not only of great importance to real-time flood management and flood forecasting but also support the upcoming Surface Water and Ocean Topography (SWOT) mission that will routinely provide water levels and slopes with higher precision around the globe.

  15. Assessment of surface solar irradiance derived from real-time modelling techniques and verification with ground-based measurements

    Science.gov (United States)

    Kosmopoulos, Panagiotis G.; Kazadzis, Stelios; Taylor, Michael; Raptis, Panagiotis I.; Keramitsoglou, Iphigenia; Kiranoudis, Chris; Bais, Alkiviadis F.

    2018-02-01

    This study focuses on the assessment of surface solar radiation (SSR) based on operational neural network (NN) and multi-regression function (MRF) modelling techniques that produce instantaneous (in less than 1 min) outputs. Using real-time cloud and aerosol optical properties inputs from the Spinning Enhanced Visible and Infrared Imager (SEVIRI) on board the Meteosat Second Generation (MSG) satellite and the Copernicus Atmosphere Monitoring Service (CAMS), respectively, these models are capable of calculating SSR in high resolution (1 nm, 0.05°, 15 min) that can be used for spectrally integrated irradiance maps, databases and various applications related to energy exploitation. The real-time models are validated against ground-based measurements of the Baseline Surface Radiation Network (BSRN) in a temporal range varying from 15 min to monthly means, while a sensitivity analysis of the cloud and aerosol effects on SSR is performed to ensure reliability under different sky and climatological conditions. The simulated outputs, compared to their common training dataset created by the radiative transfer model (RTM) libRadtran, showed median error values in the range -15 to 15 % for the NN that produces spectral irradiances (NNS), 5-6 % underestimation for the integrated NN and close to zero errors for the MRF technique. The verification against BSRN revealed that the real-time calculation uncertainty ranges from -100 to 40 and -20 to 20 W m-2, for the 15 min and monthly mean global horizontal irradiance (GHI) averages, respectively, while the accuracy of the input parameters, in terms of aerosol and cloud optical thickness (AOD and COT), and their impact on GHI, was of the order of 10 % as compared to the ground-based measurements. The proposed system aims to be utilized through studies and real-time applications which are related to solar energy production planning and use.

  16. 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.

  17. Soil Baiting, Rapid PCR Assay and Quantitative Real Time PCR to Diagnose Late Blight of Potato in Quarantine Programs

    Directory of Open Access Journals (Sweden)

    Touseef Hussain

    2018-05-01

    Full Text Available Phytophthora infestans (mont de Bary is a pathogen of great concern across the globe, and accurate detection is an important component in responding to the outbreaks of potential disease. Although the molecular diagnostic protocol used in regulatory programs has been evaluated but till date methods implying direct comparison has rarely used. In this study, a known area soil samples from potato fields where light blight appear every year (both A1 and A2 mating type was assayed by soil bait method, PCR assay detection and quantification of the inoculums. Suspected disease symptoms appeared on bait tubers were further confirmed by rapid PCR, inoculums were quantified through Real Time PCR, which confirms presence of P. infestans. These diagnostic methods can be highly correlated with one another. Potato tuber baiting increased the sensitivity of the assay compared with direct extraction of DNA from tuber and soil samples. Our study determines diagnostic sensitivity and specificity of the assays to determine the performance of each method. Overall, molecular techniques based on different types of PCR amplification and Real-time PCR can lead to high throughput, faster and more accurate detection method which can be used in quarantine programmes in potato industry and diagnostic laboratory.

  18. 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

  19. 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)

  20. Real-Time Measurement of Electronic Cigarette Aerosol Size Distribution and Metals Content Analysis.

    Science.gov (United States)

    Mikheev, Vladimir B; Brinkman, Marielle C; Granville, Courtney A; Gordon, Sydney M; Clark, Pamela I

    2016-09-01

    Electronic cigarette (e-cigarette) use is increasing worldwide and is highest among both daily and nondaily smokers. E-cigarettes are perceived as a healthier alternative to combustible tobacco products, but their health risk factors have not yet been established, and one of them is lack of data on aerosol size generated by e-cigarettes. We applied a real-time, high-resolution aerosol differential mobility spectrometer to monitor the evolution of aerosol size and concentration during puff development. Particles generated by e-cigarettes were immediately delivered for analysis with minimal dilution and therefore with minimal sample distortion, which is critically important given the highly dynamic aerosol/vapor mixture inherent to e-cigarette emissions. E-cigarette aerosols normally exhibit a bimodal particle size distribution: nanoparticles (11-25nm count median diameter) and submicron particles (96-175nm count median diameter). Each mode has comparable number concentrations (10(7)-10(8) particles/cm(3)). "Dry puff" tests conducted with no e-cigarette liquid (e-liquid) present in the e-cigarette tank demonstrated that under these conditions only nanoparticles were generated. Analysis of the bulk aerosol collected on the filter showed that e-cigarette emissions contained a variety of metals. E-cigarette aerosol size distribution is different from that of combustible tobacco smoke. E-cigarettes generate high concentrations of nanoparticles and their chemical content requires further investigation. Despite the small mass of nanoparticles, their toxicological impact could be significant. Toxic chemicals that are attached to the small nanoparticles may have greater adverse health effects than when attached to larger submicron particles. The e-cigarette aerosol size distribution is different from that of combustible tobacco smoke and typically exhibits a bimodal behavior with comparable number concentrations of nanoparticles and submicron particles. While vaping the e

  1. 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.

  2. 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)

  3. 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.

  4. Quantitative assessment of Azumiobodo hoyamushi distribution in the tunic of soft tunic syndrome-affected ascidian Halocynthia roretzi using real-time polymerase chain reaction.

    Science.gov (United States)

    Shin, Yun-Kyung; Nam, Ki-Woong; Park, Kwan Ha; Yoon, Jong-Man; Park, Kyung-Il

    2014-11-26

    The kinetoplastid parasite, Azumiobodo hoyamushi, is the causative agent of soft tunic syndrome (STS) in ascidians and leads to their mass mortality in Korean waters. This study was conducted to quantify A. hoyamushi density during the development of STS in the tunics of ascidians (Halocynthia roretzi) using real-time polymerase chain reaction (qPCR). The infection intensity of A. hoyamushi, as measured by qPCR, varied depending on the part of the tunic analyzed, as well as the stage of STS development. The highest infection intensity was recorded in the tunics of the siphons. The infection intensity of A. hoyamushi in the siphons was only 2.9 cell/tunic (area, 0.25 cm(2)) or 106.0 cell/gram tunic (GT) in the early phase of STS, but this value increased dramatically to 16,066 cells/tunic (0.25 cm(2)) or 617,004 cell/GT at the time of death. The number of A. hoyamushi parasites increased gradually and their distribution spread from the siphons to the other parts of the tunics. qPCR enabled the quantitation of A. hoyamushi and the results revealed that parasite density increased as STS progressed. In addition, our results suggested that the siphons might function as the portal of entry for A. hoyamushi during infection.

  5. A human fecal contamination score for ranking recreational sites using the HF183/BacR287 quantitative real-time PCR method.

    Science.gov (United States)

    Cao, Yiping; Sivaganesan, Mano; Kelty, Catherine A; Wang, Dan; Boehm, Alexandria B; Griffith, John F; Weisberg, Stephen B; Shanks, Orin C

    2018-01-01

    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 management. However, there are currently no standardized approaches for field implementation and interpretation of qPCR data. In this study, a standardized HF183/BacR287 qPCR method was combined with a water sampling strategy and a novel Bayesian weighted average approach to establish a human fecal contamination score (HFS) that can be used to prioritize sampling sites for remediation based on measured human waste levels. The HFS was then used to investigate 975 study design scenarios utilizing different combinations of sites with varying sampling intensities (daily to once per week) and number of qPCR replicates per sample (2-14 replicates). Findings demonstrate that site prioritization with HFS is feasible and that both sampling intensity and number of qPCR replicates influence reliability of HFS estimates. The novel data analysis strategy presented here provides a prescribed approach for the implementation and interpretation of human-associated HF183/BacR287 qPCR data with the goal of site prioritization based on human fecal pollution levels. In addition, information is provided for future users to customize study designs for optimal HFS performance. Published by Elsevier Ltd.

  6. Combined metabonomic and quantitative real-time PCR analyses reveal systems metabolic changes in Jurkat T-cells treated with HIV-1 Tat protein.

    Science.gov (United States)

    Liao, Wenting; Tan, Guangguo; Zhu, Zhenyu; Chen, Qiuli; Lou, Ziyang; Dong, Xin; Zhang, Wei; Pan, Wei; Chai, Yifeng

    2012-11-02

    HIV-1 Tat protein is released by infected cells and can affect bystander uninfected T cells and induce numerous biological responses which contribute to its pathogenesis. To elucidate the complex pathogenic mechanism, we conducted a comprehensive investigation on Tat protein-related extracellular and intracellular metabolic changes in Jurkat T-cells using combined gas chromatography-mass spectrometry (GC-MS), reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) and a hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-MS)-based metabonomics approach. Quantitative real-time PCR (qRT-PCR) analyses were further employed to measure expressions of several relevant enzymes together with perturbed metabolic pathways. Combined metabonomic and qRT-PCR analyses revealed that HIV-1 Tat caused significant and comprehensive metabolic changes, as represented by significant changes of 37 metabolites and 10 relevant enzymes in HIV-1 Tat-treated cells. Using MetaboAnalyst 2.0, it was found that 11 pathways (Impact-value >0.10) among the regulated pathways were acutely perturbed, including sphingolipid metabolism, glycine, serine and threonine metabolism, pyruvate metabolism, inositol phosphate metabolism, arginine and proline metabolism, citrate cycle, phenylalanine metabolism, tryptophan metabolism, pentose phosphate pathway, glycerophospholipid metabolism, glycolysis or gluconeogenesis. These results provide metabolic evidence of the complex pathogenic mechanism of HIV-1 Tat protein as a "viral toxin", and would help obligate Tat protein as "an important target" for therapeutic intervention and vaccine development.

  7. 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.

  8. Real-time measurement of blood pressure with Nexfin in a patient with thalidomide-related phocomelia.

    Science.gov (United States)

    Earle, Rosie; Vaghadia, Himat; Shanahan, Enda; Tang, Raymond; Sawka, Andrew

    2016-11-01

    We report the novel application of photoplethysmographic technology with the Nexfin HD monitor for real-time measurement of blood pressure (BP) in a patient with tetraamelia. The patient was a 58-year-old man with tetraamelia secondary to thalidomide exposure in utero, who presented for surgical excision of a maxillary schwannoma. Because difficulty of cuff use on rudimentary limbs and failure to gain invasive arterial access due to abnormalities of limb vasculature, this population is known to pose some unique challenges for BP measurement. Nexfin may offer an alternative noninvasive method to detect BP in patients with phocomelia during the perioperative period. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Development of real-time measurement of methanol-concentration in polymer electrolyte membrane using a local NMR sensor

    International Nuclear Information System (INIS)

    Ogawa, Kuniyasu; Ito, Kohei; Haishi, Tomoyuki

    2007-01-01

    A real-time sensor to measure methanol concentration in polymer electrolyte membrane (PEM) was developed for reducing methanol cross-over in Direct Methanol Fuel Cell (DMFC). The principle of the methanol sensor is based on the chemical shift of CH and OH species under high magnetic field. The sensor consists of a planar surface coil of 1.3 mm outside diameter. NMR signal from PEM being exposed to CH3OH solvent was measured using NMR sensor. Time-dependence changes of methanol concentration in PEM were obtained from analyzing spectrum of NMR signal. (author)

  10. 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

  11. Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

    Science.gov (United States)

    Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

    2014-08-01

    Primary biological aerosol particles (PBAPs) can contribute significantly to the coarse particle burden in many environments. PBAPs can thus influence climate and precipitation systems as cloud nuclei and can spread disease to humans, animals, and plants. Measurement data and techniques for PBAPs in natural environments at high time- and size resolution are, however, sparse, and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in southwestern Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of a waveband integrated bioaerosol sensor (WIBS-4) with a ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behavior, with increased fluorescent bioparticle concentrations at night, when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each was correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multimodal distributions turning into a broad featureless single mode after averaging, and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent

  12. [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.

  13. Applications of bulk measurement techniques for the near-real-time accounting system at the BNFP

    International Nuclear Information System (INIS)

    Ellis, J.H.

    1981-11-01

    Nuclear materials accountancy at the Barnwell Nuclear Fuel Plant (BNFP) is based primarily on bulk measurement of aqueous solutions containing uranium, plutonium, and fission products. Since 1973, Allied-General Nuclear Services (AGNS) has been adapting volume measurement and measurement control techniques at the various key measurement points within the plant. Starting in 1977, AGNS has been incorporating these measurement activities into a computerized nuclear materials control and accounting system (CNMCAS). This paper presents the major features of the measurement systems and describes the results of plant-scale testing of the system using unirradiated natural uranium. The results of these tests indicate that total uncertainties of about 0.2% of throughput (2 sigma level) can be achieved for conventional accounting and about 2.0% of hold-up can be achieved for in-process inventory estimates. These results are based on measurement of almost 500 MTU of throughput over 130 operating days

  14. Real-time monitoring of emissions with traffic data, simulation and air quality measurements

    NARCIS (Netherlands)

    Klunder, G.A.; Wilmink, I.R.

    2009-01-01

    This paper investigates the possibility to decide when to apply a (dynamic) traffic management measure to improve the air quality or reduce CO2 emissions, based on a limited set of (measured) data. It is expected that a combination of monitoring and modeling is needed for reliable air quality

  15. A dynamic approach to real-time performance measurement in design projects

    DEFF Research Database (Denmark)

    Skec, Stanko; Cash, Philip; Storga, Mario

    2017-01-01

    Recent developments in engineering design management point to the need for more dynamic, fine-grain measurement approaches able to deal with multi-dimensional, cross-level process performance in product design. Thus, this paper proposes a new approach to the measurement and management of individu...

  16. Real-Time Measurements and Modelling on Dynamic Behaviour of SonoVue Bubbles Based on Light Scattering Technology

    International Nuclear Information System (INIS)

    Juan, Tu; Rongjue, Wei; Guan, J. F.; Matula, T. J.; Crum, L. A.

    2008-01-01

    The dynamic behaviour of SonoVue microbubbles, a new generation ultrasound contrast agent, is investigated in real time with light scattering method. Highly diluted SonoVue microbubbles are injected into a diluted gel made of xanthan gum and water. The responses of individual SonoVue bubbles to driven ultrasound pulses are measured. Both linear and nonlinear bubble oscillations are observed and the results suggest that SonoVue microbubbles can generate strong nonlinear responses. By fitting the experimental data of individual bubble responses with Sarkar's model, the shell coating parameter of the bubbles and dilatational viscosity is estimated to be 7.0 nm·s·Pa

  17. Characteristic evaluation of a real-time silicon dosimeter and measurement of entrance surface dose at radiography

    International Nuclear Information System (INIS)

    Fujibuchi, Toshiou; Kato, Hideyuki; Hashimoto, Masatoshi; Abe, Yukinao; Kikawa, Takashi

    2006-01-01

    It is important to grasp how much radiation exposure has occurred through radiation diagnosis, in respect to patient explanations and radiation protection. In this examination, we used a patient skin dosimeter (PSD) that measures entrance surface dose (ESD) in real time using a fluoroscopy procedure. The PSD has the ability to display results beginning at 1 μGy. We focused our attention on the X-ray detectability of the PSD, and performed a representative evaluation with the X-ray equipment. We measured ESD under various radiographic parameters at our facility. Although the measurements were dependent on energy, we were able to measure ESD to within an accuracy of about a 5% error by putting a calibration value on energy. The PSD can measure ESD easily without requiring preparation. It is important to be aware of the exposure dose to the radiation staff, and the PSD is a very effective radiation dose-measuring tool when daily business is active. (author)

  18. Can We Trust Real Time Measurements of Lung Deposited Surface Area Concentrations in Dust from Powder Nanomaterials?

    DEFF Research Database (Denmark)

    Levin, Marcus; Witschger, Olivier; Bau, Sebastien

    2016-01-01

    A comparison between various methods for real-time measurements of lung deposited surface area (LDSA) using spherical particles and powder dust with specific surface area ranging from 0.03 to 112 m2 g-1 was conducted. LDSA concentrations measured directly using Nanoparticle Surface Area Monitor...... gravimetrical filter measurements and specific surface areas. Measurement of LDSA showed very good correlation in measurements of spherical particles (R2 > 0.97, Ratio 1.0 to 1.04). High surface area nanomaterial powders showed a fairly reliable correlation between NSAM and Aerotrak (R2 0...... present. We conclude that there is currently insufficient reliability and comparability between methods in the measurement of LDSA concentrations. Further development is required to enable use of LDSA for reliable dose metric and regulatory enforcement of exposure....

  19. 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.

  20. 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.

  1. Real-time measurement of aerosol particle concentration at high temperatures; Hiukkaspitoisuuden reaaliaikainen mittaaminen korkeassa laempoetilassa

    Energy Technology Data Exchange (ETDEWEB)

    Keskinen, J; Hautanen, J; Laitinen, A [Tampere Univ. of Technology (Finland). Physics

    1997-10-01

    The aim of this project is to develop a new method for continuous aerosol particle concentration measurement at elevated temperatures (up to 800-1000 deg C). The measured property of the aerosol particles is the so called Fuchs surface area. This quantity is relevant for diffusion limited mass transfer to particles. The principle of the method is as follows. First, aerosol particles are charged electrically by diffusion charging process. The charging takes place at high temperature. After the charging, aerosol is diluted and cooled. Finally, aerosol particles are collected and the total charge carried by the aerosol particles is measured. Particle collection and charge measurement take place at low temperature. Benefits of this measurement method are: particles are charged in-situ, charge of the particles is not affected by the temperature and pressure changes after sampling, particle collection and charge measurement are carried out outside the process conditions, and the measured quantity is well defined. The results of this study can be used when the formation of the fly ash particles is studied. Another field of applications is the study and the development of gasification processes. Possibly, the method can also be used for the monitoring the operation of the high temperature particle collection devices. (orig.)

  2. HIV-1 viral load measurement in venous blood and fingerprick blood using Abbott RealTime HIV-1 DBS assay.

    Science.gov (United States)

    Tang, Ning; Pahalawatta, Vihanga; Frank, Andrea; Bagley, Zowie; Viana, Raquel; Lampinen, John; Leckie, Gregor; Huang, Shihai; Abravaya, Klara; Wallis, Carole L

    2017-07-01

    HIV RNA suppression is a key indicator for monitoring success of antiretroviral therapy. From a logistical perspective, viral load (VL) testing using Dried Blood Spots (DBS) is a promising alternative to plasma based VL testing in resource-limited settings. To evaluate the analytical and clinical performance of the Abbott RealTime HIV-1 assay using a fully automated one-spot DBS sample protocol. Limit of detection (LOD), linearity, lower limit of quantitation (LLQ), upper limit of quantitation (ULQ), and precision were determined using serial dilutions of HIV-1 Virology Quality Assurance stock (VQA Rush University), or HIV-1-containing armored RNA, made in venous blood. To evaluate correlation, bias, and agreement, 497 HIV-1 positive adult clinical samples were collected from Ivory Coast, Uganda and South Africa. For each HIV-1 participant, DBS-fingerprick, DBS-venous and plasma sample results were compared. Correlation and bias values were obtained. The sensitivity and specificity were analyzed at a threshold of 1000 HIV-1 copies/mL generated using the standard plasma protocol. The Abbott HIV-1 DBS protocol had an LOD of 839 copies/mL, a linear range from 500 to 1×10 7 copies/mL, an LLQ of 839 copies/mL, a ULQ of 1×10 7 copies/mL, and an inter-assay SD of ≤0.30 log copies/mL for all tested levels within this range. With clinical samples, the correlation coefficient (r value) was 0.896 between DBS-fingerprick and plasma and 0.901 between DBS-venous and plasma, and the bias was -0.07 log copies/mL between DBS-fingerprick and plasma and -0.02 log copies/mL between DBS-venous and plasma. The sensitivity of DBS-fingerprick and DBS-venous was 93%, while the specificity of both DBS methods was 95%. The results demonstrated that the Abbott RealTime HIV-1 assay with DBS sample protocol is highly sensitive, specific and precise across a wide dynamic range and correlates well with plasma values. The Abbott RealTime HIV-1 assay with DBS sample protocol provides an

  3. A real-time measurement system for parameters of live biology metabolism process with fiber optics

    Science.gov (United States)

    Tao, Wei; Zhao, Hui; Liu, Zemin; Cheng, Jinke; Cai, Rong

    2010-08-01

    Energy metabolism is one of the basic life activities of cellular in which lactate, O2 and CO2 will be released into the extracellular environment. By monitoring the quantity of these parameters, the mitochondrial performance will be got. A continuous measurement system for the concentration of O2, CO2 and PH value is introduced in this paper. The system is made up of several small-sized fiber optics biosensors corresponding to the container. The setup of the system and the principle of measurement of several parameters are explained. The setup of the fiber PH sensor based on principle of light absorption is also introduced in detail and some experimental results are given. From the results we can see that the system can measure the PH value precisely suitable for cell cultivation. The linear and repeatable accuracies are 3.6% and 6.7% respectively, which can fulfill the measurement task.

  4. Application-Driven Reliability Measures and Evaluation Tool for Fault-Tolerant Real-Time Systems

    National Research Council Canada - National Science Library

    Krishna, C

    2001-01-01

    .... The measure combines graphic-theoretic concepts in evaluating the underlying reliability of the network and other means to evaluate the ability of the network to support interprocessor traffic...

  5. 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

  6. Extreme Design Loads Calibration of Offshore Wind Turbine Blades through Real Time Measurements

    DEFF Research Database (Denmark)

    Natarajan, Anand; Vesth, Allan; Lamata, Rebeca Rivera

    2014-01-01

    Blade Root flap and Edge moments are measured on the blades of a 3.6MW offshore wind turbine in normal operation. Ten minute maxima of the measurements are sampled to determine the extreme blade root flap moment, edge moment and resultant moment over six month duration. A random subset of the mea......Blade Root flap and Edge moments are measured on the blades of a 3.6MW offshore wind turbine in normal operation. Ten minute maxima of the measurements are sampled to determine the extreme blade root flap moment, edge moment and resultant moment over six month duration. A random subset...... of the measurements over a week is taken as input to stochastic load extrapolation whereby the one year extrapolated design extreme is obtained, which are then compared with the maximum extremes obtained from direct measurements over a six month period to validate the magnification in the load levels for the blade...... root flap moment, edge moment obtained by extrapolation. The validation yields valuable information on prescribing the slope of the local extrapolation curve at each mean wind speed. As an alternative to determining the contemporaneous loads for each primary extrapolated load, the blade root resultant...

  7. Uncertainty modelling of real-time observation of a moving object: photogrammetric measurements

    Science.gov (United States)

    Ulrich, Thomas

    2015-04-01

    Photogrametric systems are widely used in the field of industrial metrology to measure kinematic tasks such as tracking robot movements. In order to assess spatiotemporal deviations of a kinematic movement, it is crucial to have a reliable uncertainty of the kinematic measurements. Common methods to evaluate the uncertainty in kinematic measurements include approximations specified by the manufactures, various analytical adjustment methods and Kalman filters. Here a hybrid system estimator in conjunction with a kinematic measurement model is applied. This method can be applied to processes which include various types of kinematic behaviour, constant velocity, variable acceleration or variable turn rates. Additionally, it has been shown that the approach is in accordance with GUM (Guide to the Expression of Uncertainty in Measurement). The approach is compared to the Kalman filter using simulated data to achieve an overall error calculation. Furthermore, the new approach is used for the analysis of a rotating system as this system has both a constant and a variable turn rate. As the new approach reduces overshoots it is more appropriate for analysing kinematic processes than the Kalman filter. In comparison with the manufacturer’s approximations, the new approach takes account of kinematic behaviour, with an improved description of the real measurement process. Therefore, this approach is well-suited to the analysis of kinematic processes with unknown changes in kinematic behaviour.

  8. Development of a method for measuring femoral torsion using real-time ultrasound

    International Nuclear Information System (INIS)

    Hafiz, Eliza; Hiller, Claire E; Nightingale, E Jean; Eisenhuth, John P; Refshauge, Kathryn M; Nicholson, Leslie L; Clarke, Jillian L; Grimaldi, Alison

    2014-01-01

    Excessive femoral torsion has been associated with various musculoskeletal and neurological problems. To explore this relationship, it is essential to be able to measure femoral torsion in the clinic accurately. Computerized tomography (CT) and magnetic resonance imaging (MRI) are thought to provide the most accurate measurements but CT involves significant radiation exposure and MRI is expensive. The aim of this study was to design a method for measuring femoral torsion in the clinic, and to determine the reliability of this method. Details of design process, including construction of a jig, the protocol developed and the reliability of the method are presented. The protocol developed used ultrasound to image a ridge on the greater trochanter, and a customized jig placed on the femoral condyles as reference points. An inclinometer attached to the customized jig allowed quantification of the degree of femoral torsion. Measurements taken with this protocol had excellent intra- and inter-rater reliability (ICC 2,1  = 0.98 and 0.97, respectively). This method of measuring femoral torsion also permitted measurement of femoral torsion with a high degree of accuracy. This method is applicable to the research setting and, with minor adjustments, will be applicable to the clinical setting. (paper)

  9. Overview report of the international workshop on the near-real-time accountancy measure

    International Nuclear Information System (INIS)

    Gupta, D.

    1983-07-01

    After establishing the process and accountancy data-base for a 1000 t HM/a reference reprocessing facility, the workshop developed simulation models for the sequential generation of data for throughput and inventory of plutonium in the process material balance area (MBA). A well defined set of boundary conditions and parameter values for measurement uncertainties and loss patterns was established, on the basis of which a number of sequential statistical test procedures was evaluated. One important condition for the application of the NRTA measure was the stipulation that routinely measured Pu inventories in process tanks only, would be used, since more than 95% of Pu inventories in the process MBA are in these tanks. About 12 kg of Pu, expected to be the normal inventory in six pulse columns, was assumed to be constant. In spite of the simplifications made and the fact that mainly simulated data were used, these investigations permit the conclusion that the NRTA measure provides a greater sensitivity in terms of the amounts which can be detected and the timeliness of detection, than the conventional material accountancy. Since measurements are restricted to process tanks only, routinely available measurement techniques can be used. (orig.) [de

  10. Comparison of real-time and quantitative polymerase chain reaction assays in detection of cytomegalovirus DNA in clinical specimens

    International Nuclear Information System (INIS)

    Gokahmetoglu, S.; Deniz, E.

    2007-01-01

    To compare the real-time (RT) and qualitative (Q) polymerase chain reaction (PCR) assays for detection of Cytomegalovirus (CMV) DNA. The study took place in the Department of Microbiology, Erciyes University, Kayseri and in Iontek Laboratory, Istanbul, Turkey, from August to December 2006. One hundred and seven clinical specimens from 67 patients were included in the study. Cytomegalovirus DNA was investigated using RT-PCR kit (Fluorion Iontek, Turkey) and Q-PCR kit (Fluorion Iontek, Turkey). Deoxyribonucleic acid sequencing was applied to the samples that yielded discrepant results in both assays. Mac Nema's Chi Square test was used for statistical analysis. Of the specimens, 27 were found positive with both assays: 9 with only RT-PCR, and 11 with only Q-PCR assay. Both assays were found negative in 60 of the specimens. There was a good agreement between the 2 assays in 87(81.3%) of the specimens. There was no statistical significant difference between the assays (p>0.05). Two of the 11 samples that RT-PCR negative Q-PCR positive, and 3 of 9 samples that RT-PCR positive Q-PCR negative were found to be CMV DNA positive by DNA sequencing. A good level of concordance between RT-PCR and Q-PCR assays for CMV DNA detection has been found. (author)

  11. Quantitative evaluation of alternatively spliced mRNA isoforms by label-free real-time plasmonic sensing.

    Science.gov (United States)

    Huertas, César S; Carrascosa, L G; Bonnal, S; Valcárcel, J; Lechuga, L M

    2016-04-15

    Alternative splicing of mRNA precursors enables cells to generate different protein outputs from the same gene depending on their developmental or homeostatic status. Its deregulation is strongly linked to disease onset and progression. Current methodologies for monitoring alternative splicing demand elaborate procedures and often present difficulties in discerning between closely related isoforms, e.g. due to cross-hybridization during their detection. Herein, we report a general methodology using a Surface Plasmon Resonance (SPR) biosensor for label-free monitoring of alternative splicing events in real-time, without any cDNA synthesis or PCR amplification requirements. We applied this methodology to RNA isolated from HeLa cells for the quantification of alternatively spliced isoforms of the Fas gene, involved in cancer progression through regulation of programmed cell death. We demonstrate that our methodology is isoform-specific, with virtually no cross-hybridization, achieving limits of detection (LODs) in the picoMolar (pM) range. Similar results were obtained for the detection of the BCL-X gene mRNA isoforms. The results were independently validated by RT-qPCR, with excellent concordance in the determination of isoform ratios. The simplicity and robustness of this biosensor technology can greatly facilitate the exploration of alternative splicing biomarkers in disease diagnosis and therapy. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Development of Real-Time Measurement of Effective Dose for High Dose Rate Neutron Fields

    International Nuclear Information System (INIS)

    Braby, L. A.; Reece, W. D.; Hsu, W. H.

    2003-01-01

    Studies of the effects of low doses of ionizing radiation require sources of radiation which are well characterized in terms of the dose and the quality of the radiation. One of the best measures of the quality of neutron irradiation is the dose mean lineal energy. At very low dose rates this can be determined by measuring individual energy deposition events, and calculating the dose mean of the event size. However, at the dose rates that are normally required for biology experiments, the individual events can not be separated by radiation detectors. However, the total energy deposited in a specified time interval can be measured. This total energy has a random variation which depends on the size of the individual events, so the dose mean lineal energy can be calculated from the variance of repeated measurements of the energy deposited in a fixed time. We have developed a specialized charge integration circuit for the measurement of the charge produced in a small ion chamber in typical neutron irradiation experiments. We have also developed 4.3 mm diameter ion chambers with both tissue equivalent and carbon walls for the purpose of measuring dose mean lineal energy due to all radiations and due to all radiations except neutrons, respectively. By adjusting the gas pressure in the ion chamber, it can be made to simulate tissue volumes from a few nanometers to a few millimeters in diameter. The charge is integrated for 0.1 seconds, and the resulting pulse height is recorded by a multi channel analyzer. The system has been used in a variety of photon and neutron radiation fields, and measured values of dose and dose mean lineal energy are consistent with values extrapolated from measurements made by other techniques at much lower dose rates. It is expected that this technique will prove to be much more reliable than extrapolations from measurements made at low dose rates because these low dose rate exposures generally do not accurately reproduce the attenuation and

  13. Development of Real-Time Measurement of Effective Dose for High Dose Rate Neutron Fields

    CERN Document Server

    Braby, L A; Reece, W D

    2003-01-01

    Studies of the effects of low doses of ionizing radiation require sources of radiation which are well characterized in terms of the dose and the quality of the radiation. One of the best measures of the quality of neutron irradiation is the dose mean lineal energy. At very low dose rates this can be determined by measuring individual energy deposition events, and calculating the dose mean of the event size. However, at the dose rates that are normally required for biology experiments, the individual events can not be separated by radiation detectors. However, the total energy deposited in a specified time interval can be measured. This total energy has a random variation which depends on the size of the individual events, so the dose mean lineal energy can be calculated from the variance of repeated measurements of the energy deposited in a fixed time. We have developed a specialized charge integration circuit for the measurement of the charge produced in a small ion chamber in typical neutron irradiation exp...

  14. Development of real-time and quantitative monitoring of thrombus formation in an extracorporeal centrifugal blood pump

    Science.gov (United States)

    Sakota, Daisuke; Fujiwara, Tatsuki; Ohuchi, Katsuhiro; Kuwana, Katsuyuki; Yamazaki, Hiroyuki; Kosaka, Ryo; Maruyama, Osamu

    2018-02-01

    We developed an optical detector of thrombus formed on the pivot bearing of an extracorporeal centrifugal blood pump (MERA HCF-MP23; Senko Medical Instrument Mfg. Co., Ltd., Tokyo, Japan) which is frequently used for long-term extracorporeal circulation support to bridge to an implantable artificial heart, which in turn is used for bridge to heart transplantation in Japan. In this study, we investigated the quantitative performance of the thrombus formation in acute animal experiments. A total of three experiments of extracorporeal left ventricular assist using Japanese specific pathogen-free pigs were conducted. The optical fibers were set in the pump driver unit. The incident light at nearinfrared wavelength aiming at the pivot bearing and the resulting scattered light were guided to respective fibers. The detected signal was analyzed to obtain thrombus formation level (TFL) calculated by a specially developed software. When the increase in TFL was confirmed, the pump was exchanged and the extracorporeal circulation was restarted. The number of pump exchanges were four times at each experiment so a total of twelve pumps were evaluated. 3-dimentional data surrounding the pivot bearing and the adhered thrombus was captured by a 3-dimantional surface measurement system to calculate the thrombus surface area (TSA) formed on the pivot bearing. As a result, the correlation coefficient between TFL and TSA was 0.878. The accuracy of TSA estimated by the optical detector was 3.6+/-2.3 mm2. This was small enough to not have the pump exchanged in clinical judgement. The developed detector would be useful for optimal anti-coagulation management.

  15. Nonintrusive performance measurement of a gas turbine engine in real time

    Science.gov (United States)

    DeSilva, Upul P.; Claussen, Heiko

    2017-08-29

    Performance of a gas turbine engine is monitored by computing a mass flow rate through the engine. Acoustic time-of-flight measurements are taken between acoustic transmitters and receivers in the flow path of the engine. The measurements are processed to determine average speeds of sound and gas flow velocities along those lines-of-sound. A volumetric flow rate in the flow path is computed using the gas flow velocities together with a representation of the flow path geometry. A gas density in the flow path is computed using the speeds of sound and a measured static pressure. The mass flow rate is calculated from the gas density and the volumetric flow rate.

  16. Real-time ultrafast dynamics of dense, hot matter measured by pump-probe Doppler spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lad, Amit D; Mondal, S; Narayanan, V; Ahmed, Saima; Kumar, G Ravindra; Rajeev, P P; Robinson, A P L [Central Laser Facility, Rutherford-Appleton Laboratory, Chilton, Oxfordshire (United Kingdom); Pasley, J, E-mail: amitlad@tifr.res.i [Department of Physics, University of York, Heslington, York (United Kingdom)

    2010-08-01

    A detailed understanding of the critical surface motion of high intensity laser produced plasma is very crucial for understanding the interaction. We employ the two colour pump-probe technique to report the first ever femtosecond scale ultrafast dynamics measurement of the critical surface of a solid plasma produced by a relativistically intense, femtosecond pump laser beam (10{sup 18} W/cm{sup 2}, 30 fs, 800 nm) on an aluminium target. We observe the Doppler shift of a time delayed probe laser beam (10{sup 12} W/cm{sup 2}, 80 fs, 400 nm) up to delays of 30 ps. Such unravelling of dynamics has not been possible in earlier measurements, which typically used the self reflection of a powerful pump pulse. We observe time dependent red and blue shifts and measure their magnitudes to infer plasma expansion velocity and acceleration and thereby the plasma profile. Our results are very well reproduced by 1D hydrodynamic simulation (HYADES code).

  17. A study on the real-time radiation dosimetry measurement system based on optically stimulated luminescence

    International Nuclear Information System (INIS)

    Liu Yanping; Du Yanzhao; Chen Zhaoyang; Ba Weizhen; Fan Yanwei; Pan Shilie; Guo Qi

    2008-01-01

    The optically stimulated luminescent (OSL) radiation dosimeter technically surveys a wide dynamic measurement range and a high sensitivity. Optical fiber dosimeters provide capability for remote monitoring of the radiation in the locations which are difficult-to-access and hazardous. In addition, optical fiber dosimeters are immune to electrical and radio-frequency interference. In this paper, a novel remote optical fiber radiation dosimeter is described. The optical fiber dosimeter takes advantage of the charge trapping materials CaS:Ce, Sm that exhibit OSL. The measuring range of the dosimeter is from 0.1 to 100 Gy. The equipment is relatively simple and small in size, and has low power consumption. This device is suitable for measuring the space radiation dose and also can be used in high radiation dose condition and other dangerous radiation occasions. (authors)

  18. 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...

  19. 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

  20. Real-time estimation of helicopter rotor blade kinematics through measurement of rotation induced acceleration

    Science.gov (United States)

    Allred, C. Jeff; Churchill, David; Buckner, Gregory D.

    2017-07-01

    This paper presents a novel approach to monitoring rotor blade flap, lead-lag and pitch using an embedded gyroscope and symmetrically mounted MEMS accelerometers. The central hypothesis is that differential accelerometer measurements are proportional only to blade motion; fuselage acceleration and blade bending are inherently compensated for. The inverse kinematic relationships (from blade position to acceleration and angular rate) are derived and simulated to validate this hypothesis. An algorithm to solve the forward kinematic relationships (from sensor measurement to blade position) is developed using these simulation results. This algorithm is experimentally validated using a prototype device. The experimental results justify continued development of this kinematic estimation approach.

  1. Ultrasonic device for real-time sewage velocity and suspended particles concentration measurements.

    Science.gov (United States)

    Abda, F; Azbaid, A; Ensminger, D; Fischer, S; François, P; Schmitt, P; Pallarès, A

    2009-01-01

    In the frame of a technological research and innovation network in water and environment technologies (RITEAU, Réseau de Recherche et d'Innovation Technologique Eau et Environnement), our research group, in collaboration with industrial partners and other research institutions, has been in charge of the development of a suitable flowmeter: an ultrasonic device measuring simultaneously the water flow and the concentration of size classes of suspended particles. Working on the pulsed ultrasound principle, our multi-frequency device (1 to 14 MHz) allows flow velocity and water height measurement and estimation of suspended solids concentration. Velocity measurements rely on the coherent Doppler principle. A self developed frequency estimator, so called Spectral Identification method, was used and compared to the classical Pulse-Pair method. Several measurements campaigns on one wastewater collector of the French city of Strasbourg gave very satisfactory results and showed smaller standard deviation values for the Doppler frequency extracted by the Spectral Identification method. A specific algorithm was also developed for the water height measurements. It relies on the water surface acoustic impedance rupture and its peak localisation and behaviour in the collected backscattering data. This algorithm was positively tested on long time measurements on the same wastewater collector. A large part of the article is devoted to the measurements of the suspended solids concentrations. Our data analysis consists in the adaptation of the well described acoustic behaviour of sand to the behaviour of wastewater particles. Both acoustic attenuation and acoustic backscattering data over multiple frequencies are analyzed for the extrapolation of size classes and respective concentrations. Under dry weather conditions, the massic backscattering coefficient and the overall size distribution showed similar evolution whatever the measurement site was and were suggesting a global

  2. Comparison of different real time VOC measurement techniques in a ponderosa pine forest

    Directory of Open Access Journals (Sweden)

    L. Kaser

    2013-03-01

    Full Text Available Volatile organic compound (VOC mixing ratios measured by five independent instruments are compared at a forested site dominated by ponderosa pine (Pinus Ponderosa during the BEACHON-ROCS field study in summer 2010. The instruments included a Proton Transfer Reaction Time of Flight Mass Spectrometer (PTR-TOF-MS, a Proton Transfer Reaction Quadrupole Mass Spectrometer (PTR-MS, a Fast Online Gas-Chromatograph coupled to a Mass Spectrometer (GC/MS; TOGA, a Thermal Dissociation Chemical Ionization Mass Spectrometer (PAN-CIMS and a Fiber Laser-Induced Fluorescence Instrument (FILIF. The species discussed in this comparison include the most important biogenic VOCs and a selected suite of oxygenated VOCs that are thought to dominate the VOC reactivity at this particular site as well as typical anthropogenic VOCs that showed low mixing ratios at this site. Good agreement was observed for methanol, the sum of the oxygenated hemiterpene 2-methyl-3-buten-2-ol (MBO and the hemiterpene isoprene, acetaldehyde, the sum of acetone and propanal, benzene and the sum of methyl ethyl ketone (MEK and butanal. Measurements of the above VOCs conducted by different instruments agree within 20%. The ability to differentiate the presence of toluene and cymene by PTR-TOF-MS is tested based on a comparison with GC-MS measurements, suggesting a study-average relative contribution of 74% for toluene and 26% for cymene. Similarly, 2-hydroxy-2-methylpropanal (HMPR is found to interfere with the sum of methyl vinyl ketone and methacrolein (MVK + MAC using PTR-(TOF-MS at this site. A study-average relative contribution of 85% for MVK + MAC and 15% for HMPR was determined. The sum of monoterpenes measured by PTR-MS and PTR-TOF-MS was generally 20–25% higher than the sum of speciated monoterpenes measured by TOGA, which included α-pinene, β-pinene, camphene, carene, myrcene, limonene, cineole as well as other terpenes. However, this difference is consistent throughout the study

  3. Real-time line-width measurements: a new feature for reticle inspection systems

    Science.gov (United States)

    Eran, Yair; Greenberg, Gad; Joseph, Amnon; Lustig, Cornel; Mizrahi, Eyal

    1997-07-01

    The significance of line width control in mask production has become greater with the lessening of defect size. There are two conventional methods used for controlling line widths dimensions which employed in the manufacturing of masks for sub micron devices. These two methods are the critical dimensions (CD) measurement and the detection of edge defects. Achieving reliable and accurate control of line width errors is one of the most challenging tasks in mask production. Neither of the two methods cited above (namely CD measurement and the detection of edge defects) guarantees the detection of line width errors with good sensitivity over the whole mask area. This stems from the fact that CD measurement provides only statistical data on the mask features whereas applying edge defect detection method checks defects on each edge by itself, and does not supply information on the combined result of error detection on two adjacent edges. For example, a combination of a small edge defect together with a CD non- uniformity which are both within the allowed tolerance, may yield a significant line width error, which will not be detected using the conventional methods (see figure 1). A new approach for the detection of line width errors which overcomes this difficulty is presented. Based on this approach, a new sensitive line width error detector was developed and added to Orbot's RT-8000 die-to-database reticle inspection system. This innovative detector operates continuously during the mask inspection process and scans (inspects) the entire area of the reticle for line width errors. The detection is based on a comparison of measured line width that are taken on both the design database and the scanned image of the reticle. In section 2, the motivation for developing this new detector is presented. The section covers an analysis of various defect types, which are difficult to detect using conventional edge detection methods or, alternatively, CD measurements. In section 3

  4. Real-Time Measurement of Machine Efficiency during Inertia Friction Welding.

    Energy Technology Data Exchange (ETDEWEB)

    Tung, Daniel Joseph [The Ohio State Univ., Columbus, OH (United States); Mahaffey, David [Air Force Research Lab. (AFRL), Wright-Patterson AFB, OH (United States); Senkov, Oleg [Air Force Research Lab. (AFRL), Wright-Patterson AFB, OH (United States); Semiatin, Sheldon [Air Force Research Lab. (AFRL), Wright-Patterson AFB, OH (United States); Zhang, Wei [The Ohio State Univ., Columbus, OH (United States)

    2017-12-01

    Process efficiency is a crucial parameter for inertia friction welding (IFW) that is largely unknown at the present time. A new method has been developed to determine the transient profile of the IFW process efficiency by comparing the workpiece torque used to heat and deform the joint region to the total torque. Particularly, the former is measured by a torque load cell attached to the non-rotating workpiece while the latter is calculated from the deceleration rate of flywheel rotation. The experimentally-measured process efficiency for IFW of AISI 1018 steel rods is validated independently by the upset length estimated from an analytical equation of heat balance and the flash profile calculated from a finite element based thermal stress model. The transient behaviors of torque and efficiency during IFW are discussed based on the energy loss to machine bearings and the bond formation at the joint interface.

  5. 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.)

  6. Measuring the significance of pearlescence in real-time bottle forming

    Science.gov (United States)

    Nixon, J.; Menary, G.; Yan, S.

    2018-05-01

    This work examines the optical properties of polyethylene terephthalate (PET) bottles during the stretch-blow-moulding (SBM) process. PET has a relatively large process window with regards to process parameters, however if the boundaries are pushed, the resultant bottle can become insufficient for consumer requirements. One aspect of this process is the onset of pearlescence in the bottle material, where the bottle becomes opaque due to elevated stress whitening. Experimental trials were carried out using a modified free-stretch-blow machine where the deforming bottle was examined in free air. The strain values of the deformation were measured using digital image correlation (DIC) and the optical properties were measured relative to the initial amorphous PET preform. The results reveal that process parameters can significantly affect pearlescence. The detrimental level of pearlescence may be predicted therefore reducing the probability of poorly formed bottles.

  7. Toward real-time measurement of atmospheric mercury concentrations using cavity ring-down spectroscopy

    Directory of Open Access Journals (Sweden)

    X. Faïn

    2010-03-01

    Full Text Available Cavity ring-down spectroscopy (CRDS is a direct absorption technique that utilizes path lengths up to multiple kilometers in a compact absorption cell and has a significantly higher sensitivity than conventional absorption spectroscopy. This tool opens new prospects for study of gaseous elemental mercury (Hg0 because of its high temporal resolution and reduced sample volume requirements (<0.5 l of sample air. We developed a new sensor based on CRDS for measurement of (Hg0 mass concentration. Sensor characteristics include sub-ng m−3 detection limit and high temporal resolution using a frequency-doubled, tuneable dye laser emitting pulses at ~253.65 nm with a pulse repetition frequency of 50 Hz. The dye laser incorporates a unique piezo element attached to its tuning grating allowing it to tune the laser on and off the Hg0 absorption line on a pulse-to-pulse basis to facilitate differential absorption measurements. Hg0 absorption measurements with this CRDS laboratory prototype are highly linearly related to Hg0 concentrations determined by a Tekran 2537B analyzer over an Hg0 concentration range from 0.2 ng m−3 to 573 ng m−3, implying excellent linearity of both instruments. The current CRDS instrument has a sensitivity of 0.10 ng Hg0 m−3 at 10-s time resolution. Ambient-air tests showed that background Hg0 levels can be detected at low temporal resolution (i.e., 1 s, but also highlight a need for high-frequency (i.e., pulse-to-pulse differential on/off-line tuning of the laser wavelength to account for instabilities of the CRDS system and variable background absorption interferences. Future applications may include ambient Hg0 flux measurements with eddy covariance techniques, which require measurements of Hg0 concentrations with sub-ng m−3 sensitivity and sub-second time

  8. REAL-TIME STABILITY AND PROFILE COMPARISON MEASUREMENTS BETWEEN TWO DIFFERENT LTPS

    International Nuclear Information System (INIS)

    2005-01-01

    The Long Trace Profiler (LTP) is a precise angle measurement instrument, with a sensitivity and accuracy that can be in the sub-micron radian range. LTP characteristics depend on the particular LTP system schematic design, and the quality of components and assembly. The conditions of temperature, alignment, and mirror support during the measurement process vary between different laboratories, which influences significantly the test repeatability and accuracy. In this paper we introduce a direct comparison method to test the same object at the same point in the same environment at the same time by using two LTPs, which significantly increases the reliability of the comparison. A compact, portable LTP (PTLTP), which can be carried to different laboratories around the world, is used for comparison testing. Stability Comparison experiments between the LTP II at the National Synchrotron Radiation Research Center (NSRRC), and the PTLTP of Brookhaven National Laboratory (BNL) reveal significant differences in performance between the instruments. The experiment is set up so that each optical head simultaneously records both its own sample probe beam and also the probe beam from the other optical head. The two probe beams are reflected from same point on the mirror. Tests show that the stability of the PTLTP with a monolithic beam splitter is 10 times better than the stability of the LTP II which has a separated beam splitter unit. A scheme for comparing scanning measurements of a mirror is introduced. Experimental results show a significant difference between the two LTPs due mainly to distortions in the optical components inside the optical head. A new scheme is proposed for further mirror comparison scanning tests

  9. Mitochondrial DNA as a non-invasive biomarker: Accurate quantification using real time quantitative PCR without co-amplification of pseudogenes and dilution bias

    International Nuclear Information System (INIS)

    Malik, Afshan N.; Shahni, Rojeen; Rodriguez-de-Ledesma, Ana; Laftah, Abas; Cunningham, Phil

    2011-01-01

    Highlights: → Mitochondrial dysfunction is central to many diseases of oxidative stress. → 95% of the mitochondrial genome is duplicated in the nuclear genome. → Dilution of untreated genomic DNA leads to dilution bias. → Unique primers and template pretreatment are needed to accurately measure mitochondrial DNA content. -- Abstract: Circulating mitochondrial DNA (MtDNA) is a potential non-invasive biomarker of cellular mitochondrial dysfunction, the latter known to be central to a wide range of human diseases. Changes in MtDNA are usually determined by quantification of MtDNA relative to nuclear DNA (Mt/N) using real time quantitative PCR. We propose that the methodology for measuring Mt/N needs to be improved and we have identified that current methods have at least one of the following three problems: (1) As much of the mitochondrial genome is duplicated in the nuclear genome, many commonly used MtDNA primers co-amplify homologous pseudogenes found in the nuclear genome; (2) use of regions from genes such as β-actin and 18S rRNA which are repetitive and/or highly variable for qPCR of the nuclear genome leads to errors; and (3) the size difference of mitochondrial and nuclear genomes cause a 'dilution bias' when template DNA is diluted. We describe a PCR-based method using unique regions in the human mitochondrial genome not duplicated in the nuclear genome; unique single copy region in the nuclear genome and template treatment to remove dilution bias, to accurately quantify MtDNA from human samples.

  10. Development of a methodology to measure the effect of ergot alkaloids on forestomach motility using real-time wireless telemetry

    Science.gov (United States)

    Egert, Amanda; Klotz, James; McLeod, Kyle; Harmon, David

    2014-10-01

    The objectives of these experiments were to characterize rumen motility patterns of cattle fed once daily using a real-time wireless telemetry system, determine when to measure rumen motility with this system, and determine the effect of ruminal dosing of ergot alkaloids on rumen motility. Ruminally cannulated Holstein steers (n = 8) were fed a basal diet of alfalfa cubes once daily. Rumen motility was measured by monitoring real-time pressure changes within the rumen using wireless telemetry and pressure transducers. Experiment 1 consisted of three 24-h rumen pressure collections beginning immediately after feeding. Data were recorded, stored, and analyzed using iox2 software and the rhythmic analyzer. All motility variables differed (P content samples were taken on d 15. Baseline (P = 0.06) and peak (P = 0.04) pressure were lower for E+ steers. Water intake tended (P = 0.10) to be less for E+ steers the first 8 hour period after feeding. The E+ seed treatment at this dosage under thermoneutral conditions did not significantly affect rumen motility, ruminal fill, or dry matter of rumen contents.

  11. The elastic body model: a pedagogical approach integrating real time measurements and modelling activities

    International Nuclear Information System (INIS)

    Fazio, C; Guastella, I; Tarantino, G

    2007-01-01

    In this paper, we describe a pedagogical approach to elastic body movement based on measurements of the contact times between a metallic rod and small bodies colliding with it and on modelling of the experimental results by using a microcomputer-based laboratory and simulation tools. The experiments and modelling activities have been built in the context of the laboratory of mechanical wave propagation of the two-year graduate teacher education programme of Palermo's University. Some considerations about observed modifications in trainee teachers' attitudes in utilizing experiments and modelling are discussed

  12. Particle Sampling and Real Time Size Distribution Measurement in H2/O2/TEOS Diffusion Flame

    International Nuclear Information System (INIS)

    Ahn, K.H.; Jung, C.H.; Choi, M.; Lee, J.S.

    2001-01-01

    Growth characteristics of silica particles have been studied experimentally using in situ particle sampling technique from H 2 /O 2 /Tetraethylorthosilicate (TEOS) diffusion flame with carefully devised sampling probe. The particle morphology and the size comparisons are made between the particles sampled by the local thermophoretic method from the inside of the flame and by the electrostatic collector sampling method after the dilution sampling probe. The Transmission Electron Microscope (TEM) image processed data of these two sampling techniques are compared with Scanning Mobility Particle Sizer (SMPS) measurement. TEM image analysis of two sampling methods showed a good agreement with SMPS measurement. The effects of flame conditions and TEOS flow rates on silica particle size distributions are also investigated using the new particle dilution sampling probe. It is found that the particle size distribution characteristics and morphology are mostly governed by the coagulation process and sintering process in the flame. As the flame temperature increases, the effect of coalescence or sintering becomes an important particle growth mechanism which reduces the coagulation process. However, if the flame temperature is not high enough to sinter the aggregated particles then the coagulation process is a dominant particle growth mechanism. In a certain flame condition a secondary particle formation is observed which results in a bimodal particle size distribution

  13. 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

  14. 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

  15. Quantitative analysis of aortic regurgitation: real-time 3-dimensional and 2-dimensional color Doppler echocardiographic method--a clinical and a chronic animal study

    Science.gov (United States)

    Shiota, Takahiro; Jones, Michael; Tsujino, Hiroyuki; Qin, Jian Xin; Zetts, Arthur D.; Greenberg, Neil L.; Cardon, Lisa A.; Panza, Julio A.; Thomas, James D.

    2002-01-01

    BACKGROUND: For evaluating patients with aortic regurgitation (AR), regurgitant volumes, left ventricular (LV) stroke volumes (SV), and absolute LV volumes are valuable indices. AIM: The aim of this study was to validate the combination of real-time 3-dimensional echocardiography (3DE) and semiautomated digital color Doppler cardiac flow measurement (ACM) for quantifying absolute LV volumes, LVSV, and AR volumes using an animal model of chronic AR and to investigate its clinical applicability. METHODS: In 8 sheep, a total of 26 hemodynamic states were obtained pharmacologically 20 weeks after the aortic valve noncoronary (n = 4) or right coronary (n = 4) leaflet was incised to produce AR. Reference standard LVSV and AR volume were determined using the electromagnetic flow method (EM). Simultaneous epicardial real-time 3DE studies were performed to obtain LV end-diastolic volumes (LVEDV), end-systolic volumes (LVESV), and LVSV by subtracting LVESV from LVEDV. Simultaneous ACM was performed to obtain LVSV and transmitral flows; AR volume was calculated by subtracting transmitral flow volume from LVSV. In a total of 19 patients with AR, real-time 3DE and ACM were used to obtain LVSVs and these were compared with each other. RESULTS: A strong relationship was found between LVSV derived from EM and those from the real-time 3DE (r = 0.93, P <.001, mean difference (3D - EM) = -1.0 +/- 9.8 mL). A good relationship between LVSV and AR volumes derived from EM and those by ACM was found (r = 0.88, P <.001). A good relationship between LVSV derived from real-time 3DE and that from ACM was observed (r = 0.73, P <.01, mean difference = 2.5 +/- 7.9 mL). In patients, a good relationship between LVSV obtained by real-time 3DE and ACM was found (r = 0.90, P <.001, mean difference = 0.6 +/- 9.8 mL). CONCLUSION: The combination of ACM and real-time 3DE for quantifying LV volumes, LVSV, and AR volumes was validated by the chronic animal study and was shown to be clinically applicable.

  16. Real-time measurement of inhaled and exhaled cigarette smoke: Implications for dose

    Energy Technology Data Exchange (ETDEWEB)

    McGrath, Conor; Warren, Nigel; Biggs, Philip; McAughey, John, E-mail: conor_mcgrath@bat.co [British American Tobacco, Group R and D Centre, Southampton, SO15 8TL (United Kingdom)

    2009-02-01

    Inhalation of tobacco smoke aerosol is a two-step process involving puffing followed by inhalation. Measured smoke deposition efficiencies in the lung (20-70%) are greater than expected for smoke particles of 150 -- 250 nm count median diameter (CMD). Various mechanisms have been put forward to explain this enhanced deposition pattern, including coagulation, hygroscopic growth, condensation and evaporation, changes in composition, or changes in inhalation behaviour. This paper represents one of a series of studies seeking to better quantify smoke chemistry, inhalation behaviour and cumulative particle growth. The studies have been conducted to better understand smoke dosimetry and links to disease as part of a wider programme defining risk and potential harm reduction. In this study, the average CMD of inhaled smoke was 160 nm while the average CMD of exhaled smoke was 239 nm with an average growth factor of 1.5.

  17. Real-Time Detection and Measurement of Eye Features from Color Images

    Directory of Open Access Journals (Sweden)

    Diana Borza

    2016-07-01

    Full Text Available The accurate extraction and measurement of eye features is crucial to a variety of domains, including human-computer interaction, biometry, and medical research. This paper presents a fast and accurate method for extracting multiple features around the eyes: the center of the pupil, the iris radius, and the external shape of the eye. These features are extracted using a multistage algorithm. On the first stage the pupil center is localized using a fast circular symmetry detector and the iris radius is computed using radial gradient projections, and on the second stage the external shape of the eye (of the eyelids is determined through a Monte Carlo sampling framework based on both color and shape information. Extensive experiments performed on a different dataset demonstrate the effectiveness of our approach. In addition, this work provides eye annotation data for a publicly-available database.

  18. [Real-time measurement of indoor particulate matter originating from environmental tobacco smoke: a pilot study].

    Science.gov (United States)

    Invernizzi, Giovanni; Ruprecht, Ario; Mazza, Roberto; Majno, Edoardo; Rossetti, Edoardo; Paredi, Paolo; Boffi, Roberto

    2002-01-01

    Short-term measurement of suspended particulate matter has been recently made possible since the release of laser-operating portable instruments. Data of a pilot study of field evaluation of environmental tobacco smoke (ETS) with a portable instrument are reported. We analysed the concentrations of total suspended particle (TSP) and of the fine particles PM10, PM7, PM2.5 and PM1 released indoor from a single cigarette, and their levels inside smoking- and non-smoking-areas of a restaurant. The results indicate that ETS creates high level indoor particulate pollution, with concentrations of PM10 exceeding air quality standards. This kind of field evaluation could allow a more careful assessing of short-term exposure to ETS and its relevance to public health.

  19. On real-time algorithms for the location search of discontinuous conductivities with one measurement

    International Nuclear Information System (INIS)

    Hanke, Martin

    2008-01-01

    We discuss, and compare, two simple methods that provide coordinates of a point in the vicinity of one inclusion within some object with homogeneous electrical properties. In the context of nondestructive testing such an inclusion may correspond to a material defect, whereas in medicine this may correspond to a lesion in the brain, to name only two possible applications. Both methods use only one pair of voltage/current measurements on the entire boundary of the object to determine a single pair of coordinates that is considered to be close to the center of the inclusion. The first method has been proposed previously by Kwon, Seo and Yoon; the second method, called here the effective dipole method, appears to be new. We discuss limitations of the two methods and derive error bounds for the effective dipole method under realistic assumptions. Finally, we also comment on other methods to localize inclusions

  20. Real-time measurement of inhaled and exhaled cigarette smoke: Implications for dose

    International Nuclear Information System (INIS)

    McGrath, Conor; Warren, Nigel; Biggs, Philip; McAughey, John

    2009-01-01

    Inhalation of tobacco smoke aerosol is a two-step process involving puffing followed by inhalation. Measured smoke deposition efficiencies in the lung (20-70%) are greater than expected for smoke particles of 150 -- 250 nm count median diameter (CMD). Various mechanisms have been put forward to explain this enhanced deposition pattern, including coagulation, hygroscopic growth, condensation and evaporation, changes in composition, or changes in inhalation behaviour. This paper represents one of a series of studies seeking to better quantify smoke chemistry, inhalation behaviour and cumulative particle growth. The studies have been conducted to better understand smoke dosimetry and links to disease as part of a wider programme defining risk and potential harm reduction. In this study, the average CMD of inhaled smoke was 160 nm while the average CMD of exhaled smoke was 239 nm with an average growth factor of 1.5.

  1. Soft X-Ray measurements and analysis on Tokamaks in view of real-time control

    International Nuclear Information System (INIS)

    Vezinet, Didier

    2013-01-01

    This thesis focuses on measuring and interpreting the Soft X-Ray (SXR) radiation (approximately [1 keV; 15 keV]) in Tokamaks. As explained in Chapter 2, this radiation conveys information about the plasma density, temperature, magnetic equilibrium and impurity content. However, the measured data is spectrally and spatially-integrated and results from several physical phenomena affecting every ion species. Tore Supra's SXR diagnostics is based on semiconductor diodes presented in Chapter 3, along with a new gas detector successfully tested in laboratory and on Tore Supra. A new methodology for absolute spectral characterisation of photo detectors using a portable SXR tube is presented. Tomographic inversion algorithms, that grant access to reconstructions of the SXR emissivity field in a poloidal cross-section, are presented in Chapter 4. Improvements implemented on one particular algorithm are detailed with examples of application. A comparison between the position of the SXR emissivity maximum and the magnetic axis reconstructed by an equilibrium code is presented in Chapter 5. Chapter 6 presents an approach used to derive an impurity density from its SXR emissivity using the robustness of its SXR cooling factor with respect to impurity transport. The physics accounting for this robustness is studied and a first map of the domain of validity of this method is provided. Chapter 7 addresses poloidal asymmetries of the SXR emissivity field. Two types of asymmetries are presented as well as experiments conducted on ASDEX-U to verify their parametric dependences. A new type of SXR asymmetry, observed on Tore Supra is introduced. (author) [fr

  2. A Comprehensive Statistically-Based Method to Interpret Real-Time Flowing Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Keita Yoshioka; Pinan Dawkrajai; Analis A. Romero; Ding Zhu; A. D. Hill; Larry W. Lake

    2007-01-15

    With the recent development of temperature measurement systems, continuous temperature profiles can be obtained with high precision. Small temperature changes can be detected by modern temperature measuring instruments such as fiber optic distributed temperature sensor (DTS) in intelligent completions and will potentially aid the diagnosis of downhole flow conditions. In vertical wells, since elevational geothermal changes make the wellbore temperature sensitive to the amount and the type of fluids produced, temperature logs can be used successfully to diagnose the downhole flow conditions. However, geothermal temperature changes along the wellbore being small for horizontal wells, interpretations of a temperature log become difficult. The primary temperature differences for each phase (oil, water, and gas) are caused by frictional effects. Therefore, in developing a thermal model for horizontal wellbore, subtle temperature changes must be accounted for. In this project, we have rigorously derived governing equations for a producing horizontal wellbore and developed a prediction model of the temperature and pressure by coupling the wellbore and reservoir equations. Also, we applied Ramey's model (1962) to the build section and used an energy balance to infer the temperature profile at the junction. The multilateral wellbore temperature model was applied to a wide range of cases at varying fluid thermal properties, absolute values of temperature and pressure, geothermal gradients, flow rates from each lateral, and the trajectories of each build section. With the prediction models developed, we present inversion studies of synthetic and field examples. These results are essential to identify water or gas entry, to guide flow control devices in intelligent completions, and to decide if reservoir stimulation is needed in particular horizontal sections. This study will complete and validate these inversion studies.

  3. Non-contact Real-time heart rate measurements based on high speed circuit technology research

    Science.gov (United States)

    Wu, Jizhe; Liu, Xiaohua; Kong, Lingqin; Shi, Cong; Liu, Ming; Hui, Mei; Dong, Liquan; Zhao, Yuejin

    2015-08-01

    In recent years, morbidity and mortality of the cardiovascular or cerebrovascular disease, which threaten human health greatly, increased year by year. Heart rate is an important index of these diseases. To address this status, the paper puts forward a kind of simple structure, easy operation, suitable for large populations of daily monitoring non-contact heart rate measurement. In the method we use imaging equipment video sensitive areas. The changes of light intensity reflected through the image grayscale average. The light change is caused by changes in blood volume. We video the people face which include the sensitive areas (ROI), and use high-speed processing circuit to save the video as AVI format into memory. After processing the whole video of a period of time, we draw curve of each color channel with frame number as horizontal axis. Then get heart rate from the curve. We use independent component analysis (ICA) to restrain noise of sports interference, realized the accurate extraction of heart rate signal under the motion state. We design an algorithm, based on high-speed processing circuit, for face recognition and tracking to automatically get face region. We do grayscale average processing to the recognized image, get RGB three grayscale curves, and extract a clearer pulse wave curves through independent component analysis, and then we get the heart rate under the motion state. At last, by means of compare our system with Fingertip Pulse Oximeter, result show the system can realize a more accurate measurement, the error is less than 3 pats per minute.

  4. Real-time positioning technology in horizontal directional drilling based on magnetic gradient tensor measurement

    Science.gov (United States)

    Deng, Guoqing; Yao, Aiguo

    2017-04-01

    Horizontal directional drilling (HDD) technology has been widely used in Civil Engineering. The dynamic position of the drill bit during construction is one of significant facts determining the accuracy of the trajectory of HDD. A new method now has been proposed to detecting the position of drill bit by measuring the magnetic gradient tensor of the ground solenoid magnetic beacon. Compared with traditional HDD positioning technologies, this new model is much easier to apply with lower request for construction sites and higher positioning efficiency. A direct current (DC) solenoid as a magnetic dipole is placed on ground near the drill bit, and related sensors array which contains four Micro-electromechanical Systems (MEMS ) tri-axial magnetometers, one MEMS tri-axial accelerometer and one MEMS tri-axial gyroscope is set up for measuring the magnetic gradient tensor of the magnetic dipole. The related HDD positioning model has been established and simulation experiments have been carried out to verify the feasibility and reliability of the proposed method. The experiments show that this method has good positioning accuracy in horizontal and vertical direction, and totally avoid the impact of the environmental magnetic field. It can be found that the posture of the magnetic beacon will impact the remote positioning precision within valid positioning range, and the positioning accuracy is higher with longer baseline for limited space in drilling tools. The results prove that the relative error can be limited in 2% by adjusting position of the magnetic beacon, the layers of the enameled coil, the sensitive of magnetometers and the baseline distance. Conclusion can be made that this new method can be applied in HDD positioning with better effect and wider application range than traditional method.

  5. Real-Time Rain Rate Evaluation via Satellite Downlink Signal Attenuation Measurement.

    Science.gov (United States)

    Giannetti, Filippo; Reggiannini, Ruggero; Moretti, Marco; Adirosi, Elisa; Baldini, Luca; Facheris, Luca; Antonini, Andrea; Melani, Samantha; Bacci, Giacomo; Petrolino, Antonio; Vaccaro, Attilio

    2017-08-12

    We present the NEFOCAST project (named by the contraction of "Nefele", which is the Italian spelling for the mythological cloud nymph Nephele, and "forecast"), funded by the Tuscany Region, about the feasibility of a system for the detection and monitoring of precipitation fields over the regional territory based on the use of a widespread network of new-generation Eutelsat "SmartLNB" (smart low-noise block converter) domestic terminals. Though primarily intended for interactive satellite services, these devices can also be used as weather sensors, as they have the capability of measuring the rain-induced attenuation incurred by the downlink signal and relaying it on an auxiliary return channel. We illustrate the NEFOCAST system architecture, consisting of the network of ground sensor terminals, the space segment, and the service center, which has the task of processing the information relayed by the terminals for generating rain field maps. We discuss a few methods that allow the conversion of a rain attenuation measurement into an instantaneous rainfall rate. Specifically, we discuss an exponential model relating the specific rain attenuation to the rainfall rate, whose coefficients were obtained from extensive experimental data. The above model permits the inferring of the rainfall rate from the total signal attenuation provided by the SmartLNB and from the link geometry knowledge. Some preliminary results obtained from a SmartLNB installed in Pisa are presented and compared with the output of a conventional tipping bucket rain gauge. It is shown that the NEFOCAST sensor is able to track the fast-varying rainfall rate accurately with no delay, as opposed to a conventional gauge.

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

    DEFF Research Database (Denmark)

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

    2015-01-01

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

  7. 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...

  8. Real-time black carbon emission factor measurements from light duty vehicles.

    Science.gov (United States)

    Forestieri, Sara D; Collier, Sonya; Kuwayama, Toshihiro; Zhang, Qi; Kleeman, Michael J; Cappa, Christopher D

    2013-11-19

    Eight light-duty gasoline low emission vehicles (LEV I) were tested on a Chassis dynamometer using the California Unified Cycle (UC) at the Haagen-Smit vehicle test facility at the California Air Resources Board in El Monte, CA during September 2011. The UC includes a cold start phase followed by a hot stabilized running phase. In addition, a light-duty gasoline LEV vehicle and ultralow emission vehicle (ULEV), and a light-duty diesel passenger vehicle and gasoline direct injection (GDI) vehicle were tested on a constant velocity driving cycle. A variety of instruments with response times ≥0.1 Hz were used to characterize how the emissions of the major particulate matter components varied for the LEVs during a typical driving cycle. This study focuses primarily on emissions of black carbon (BC). These measurements allowed for the determination of BC emission factors throughout the driving cycle, providing insights into the temporal variability of BC emission factors during different phases of a typical driving cycle.

  9. Drilling electrode for real-time measurement of electrical impedance in bone tissues.

    Science.gov (United States)

    Dai, Yu; Xue, Yuan; Zhang, Jianxun

    2014-03-01

    In order to prevent possible damages to soft tissues, reliable monitoring methods are required to provide valuable information on the condition of the bone being cut. This paper describes the design of an electrical impedance sensing drill developed to estimate the relative position between the drill and the bone being drilled. The two-electrode method is applied to continuously measure the electrical impedance during a drill feeding movement: two copper wire brushes are used to conduct electricity in the rotating drill and then the drill is one electrode; a needle is inserted into the soft tissues adjacent to the bone being drilled and acts as another electrode. Considering that the recorded electrical impedance is correlated with the insertion depth of the drill, we theoretically calculate the electrode-tissue contact impedance and prove that the rate of impedance change varies considerably when the drill bit crosses the boundary between two different bone tissues. Therefore, the rate of impedance change is used to determine whether the tip of the drill is located in one of cortical bone, cancellous bone, and cortical bone near a boundary with soft tissue. In vitro experiments in porcine thoracic spines were performed to demonstrate the feasibility of the impedance sensing drill. The experimental results indicate that the drill, used with the proposed data-processing method, can provide accurate and reliable breakthrough detection in the bone-drilling process.

  10. Measuring Sea Level Rise-Induced Shoreline Changes and Inundation in Real Time

    Science.gov (United States)

    Shilling, F.; Waetjen, D.; Grijalva, E.

    2016-12-01

    We describe a method to monitor shoreline inundation and changes in response to sea level rise (SLR) using a network of time-lapse cameras. We found for coastal tidal marshes that this method was sensitive to vertical changes in sea level of 20 cm has occurred in the San Francisco Bay and other US coastal areas and is likely to rise by another 30-45 cm by mid-century, which will flood and erode many coastal ecosystems, highways, and urban areas. This rapid degree of rise means that it is imperative to co-plan for natural and built systems. Many public facilities are adjacent to shoreline ecosystems, which both protect infrastructure from wave and tide energy and are home to regulated species and habitats. Accurate and timely information about the actual extent of SLR impacts to shorelines will be critical during built-system adaptation. Currently, satellite-sourced imagery cannot provide the spatial or temporal resolution necessary to investigate fine-scale shoreline changes, leaving a gap between predictive models and knowing how, where and when these changes are occurring. The method described is feasible for near-term (1 to 10 years) to long-term application and can be used for measuring fine-resolution shoreline changes (organize photographs that could be combined with related external data (e.g., gauged water levels) to create an information mashup. This information could be used to validate models predicting shoreline inundation and loss, inform SLR-adaptation planning, and to visualize SLR impacts to the public.

  11. Advancements in the Interferometric Measurements of Real Time Finishing Birefringent Filter's Crystal Plates

    International Nuclear Information System (INIS)

    Gan, Ma; Kushtal, Gi; Skomorovsky, Vi; Domyshev, Gn; Sadokhin, Vp

    2006-01-01

    The finishing of birefringent plates consists of two processes: polishing and evaluation of a surface, which have been performed separately till now. The purpose of this work is achieving of high accuracy of the evaluation and machining of the plane-parallel plates from birefringent crystals, in particular of crystal plates of birefringent filters during their finishing. The developed process combines evaluation and polishing in an interactive way. We have found modes of treatment, shape of polisher, have designed interferometer, with a mirror arranged in polisher. Visual checking of optical thickness comparatively with reference plate was carried out using the interference fringes of equal birefringence, and checking of an optical wedge - by interference rings of an equal inclination. The automated processing of TV camera interference fringes was impossible, because of gaps of interference fringes on polishing cells above the mirror. Therefore a special software was developed for processing of a complex fringe pattern interferogram. Software FastInterf uses furrier analysis technique which allows to process an interferogram with multiply gaps. Interferograms are registered by a high resolution TV camera (1280 x1024). Automatic processing of a fringe interferogram using FastInterf software takes less then one second. The influence of gaps is excluded, and the flat field is taken into account. Software provides full 3D surface and wavefront maps. Aberration analysis of a wavefront gives information on thickness of a plate comparatively with a reference one, optical wedge of plate and azimuth of an inclination of wave front. Moreover, software provides a control of surface quality. The measuring device, features of the software are described and process of interferometric evaluation during polishing is illustrated

  12. Advancements in the Interferometric Measurements of Real Time Finishing Birefringent Filter's Crystal Plates

    Energy Technology Data Exchange (ETDEWEB)

    Gan, Ma [State Optical Institute, Birzhevaya linia, 12 St. Petersburg (Russian Federation); Kushtal, Gi; Skomorovsky, Vi; Domyshev, Gn; Sadokhin, Vp [Institute of Solar-Terrestrial Physics Siberian Branch of Russian Academy of Sciences, 126 Lermontova Str., PO 4026, 664033, Irkutsk (Russian Federation)

    2006-10-15

    The finishing of birefringent plates consists of two processes: polishing and evaluation of a surface, which have been performed separately till now. The purpose of this work is achieving of high accuracy of the evaluation and machining of the plane-parallel plates from birefringent crystals, in particular of crystal plates of birefringent filters during their finishing. The developed process combines evaluation and polishing in an interactive way. We have found modes of treatment, shape of polisher, have designed interferometer, with a mirror arranged in polisher. Visual checking of optical thickness comparatively with reference plate was carried out using the interference fringes of equal birefringence, and checking of an optical wedge - by interference rings of an equal inclination. The automated processing of TV camera interference fringes was impossible, because of gaps of interference fringes on polishing cells above the mirror. Therefore a special software was developed for processing of a complex fringe pattern interferogram. Software FastInterf uses furrier analysis technique which allows to process an interferogram with multiply gaps. Interferograms are registered by a high resolution TV camera (1280 x1024). Automatic processing of a fringe interferogram using FastInterf software takes less then one second. The influence of gaps is excluded, and the flat field is taken into account. Software provides full 3D surface and wavefront maps. Aberration analysis of a wavefront gives information on thickness of a plate comparatively with a reference one, optical wedge of plate and azimuth of an inclination of wave front. Moreover, software provides a control of surface quality. The measuring device, features of the software are described and process of interferometric evaluation during polishing is illustrated.

  13. Automated on-line L-edge measurement of SNM concentration for near-real-time accounting

    International Nuclear Information System (INIS)

    Russo, P.A.; Marks, T. Jr.; Stephens, M.M.; Hsue, S.T.; Baker, A.L.; Cobb, D.D.

    1982-01-01

    The L-edge densitometer developed at Los Alamos National Laboratory has been modified, tested, and demonstrated for on-line assay of special nuclear material concentration in flowing solution streams. The demonstration was part of a larger demonstration of near-real-time nuclear materials accounting during a continuous, week-long, cold operation of the Allied General Nuclear Services facility in Barnwell, South Carolina. The L-edge data were automatically analyzed and the results were transmitted to the materials accounting computer once every 5.5 min for the duration of the cold run. This report compares the results of the L-edge analyses with the delayed results obtained from destructive analysis of samples withdrawn from the same process line. Comparisons are also made with the results obtained in near real time from an automated process control instrument installed in series with the L-edge densitometer. The performance of the L-edge instrument was reliable throughout the continous operation. The assay precision was consistent with that predicted by the counting statistics of the measurement. The results of the L-edge assays show good agreement with those of the destructive assays. A gradually varying discrepancy (of a few percent) between the L-edge and the process control results remains unexplained. 9 figures

  14. Automated on-line L-edge measurement of SNM concentration for near-real-time accounting

    International Nuclear Information System (INIS)

    Russo, P.A.; Marks, T. Jr.; Stephens, M.M.; Baker, A.L.; Cobb, D.D.

    1982-09-01

    An L-edge densitometer has been modified, tested, and demonstrated for on-line assay of special nuclear material concentration in flowing solution streams. The demonstration was part of a larger demonstration of near-real-time nuclear materials accounting during a continuous, week-long, cold operation of the Barnwell facility. The L-edge data were automatically analyzed and the results were transmitted to the materials accounting computer once every 5.5 min for the duration of the cold run. This report compares the results of the L-edge analyses with the delayed results obtained from destructive analysis of samples withdrawn from the same process line. Comparisons are also made with the results obtained in near real time from an automated process control instrument installed in series with the L-edge densitometer. The performance of the L-edge instrument was reliable throughout the continuous operation. The assay precision was consistent with that predicted by the counting statistics of the measurement. The results of the L-edge assays show good agreement with those of the destructive assays. A gradually varying discrepancy (of a few percent) between the L-edge and the process control results remains unexplained

  15. Development of a Rapid Real-Time PCR Assay for Quantitation of Pneumocystis carinii f. sp. Carinii

    DEFF Research Database (Denmark)

    Larsen, Hans Henrik; Kovacs, Joseph A; Stock, Frida

    2002-01-01

    A method for reliable quantification of Pneumocystis carinii in research models of P. carinii pneumonia (PCP) that is more convenient and reproducible than microscopic enumeration of organisms would greatly facilitate investigations of this organism. We developed a rapid quantitative touchdown (QTD......) PCR assay for detecting P. carinii f. sp. carinii, the subspecies of P. carinii commonly used in research models of PCP. The assay was based on the single-copy dihydrofolate reductase gene and was able to detect ... 6 log values for standards containing > or =5 copies/tube. Application of the assay to a series of 10-fold dilutions of P. carinii organisms isolated from rat lung demonstrated that it was reproducibly quantitative over 5 log values (r = 0.99). The assay was applied to a recently reported in vitro...

  16. Development of a Rapid Real-Time PCR Assay for Quantitation of Pneumocystis carinii f. sp. Carinii

    DEFF Research Database (Denmark)

    Larsen, Hans Henrik; Kovacs, Joseph A; Stock, Frida

    2002-01-01

    6 log values for standards containing > or =5 copies/tube. Application of the assay to a series of 10-fold dilutions of P. carinii organisms isolated from rat lung demonstrated that it was reproducibly quantitative over 5 log values (r = 0.99). The assay was applied to a recently reported in vitro...... axenic cultivation system for P. carinii and confirmed our microscopy findings that no organism multiplication had occurred during culture. For all cultures analyzed, QTD PCR assays showed a decrease in P. carinii DNA that exceeded the expected decrease due to dilution of the inoculum upon transfer......A method for reliable quantification of Pneumocystis carinii in research models of P. carinii pneumonia (PCP) that is more convenient and reproducible than microscopic enumeration of organisms would greatly facilitate investigations of this organism. We developed a rapid quantitative touchdown (QTD...

  17. Development of a Rapid Real-Time PCR Assay for Quantitation of Pneumocystis carinii f. sp. Carinii

    DEFF Research Database (Denmark)

    Larsen, Hans Henrik; Kovacs, Joseph A; Stock, Frida

    2002-01-01

    ) PCR assay for detecting P. carinii f. sp. carinii, the subspecies of P. carinii commonly used in research models of PCP. The assay was based on the single-copy dihydrofolate reductase gene and was able to detect r = 0.99) over...... 6 log values for standards containing > or =5 copies/tube. Application of the assay to a series of 10-fold dilutions of P. carinii organisms isolated from rat lung demonstrated that it was reproducibly quantitative over 5 log values (r = 0.99). The assay was applied to a recently reported in vitro....... In conclusion, a rapid, sensitive, and reproducible quantitative PCR assay for P. carinii f. sp. carinii has been developed and is applicable to in vivo as well as in vitro systems. The assay should prove useful for conducting studies in which quantification of organism burden or growth assessment is critical...

  18. Selection of Suitable Internal Control Genes for Accurate Normalization of Real-Time Quantitative PCR Data of Buffalo (Bubalus bubalis) Blastocysts Produced by SCNT and IVF.

    Science.gov (United States)

    Sood, Tanushri Jerath; Lagah, Swati Viviyan; Sharma, Ankita; Singla, Suresh Kumar; Mukesh, Manishi; Chauhan, Manmohan Singh; Manik, Radheysham; Palta, Prabhat

    2017-10-01

    We evaluated the suitability of 10 candidate internal control genes (ICGs), belonging to different functional classes, namely ACTB, EEF1A1, GAPDH, HPRT1, HMBS, RPS15, RPS18, RPS23, SDHA, and UBC for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of blastocyst-stage buffalo embryos produced by hand-made cloning and in vitro fertilization (IVF). Total RNA was isolated from three pools, each of cloned and IVF blastocysts (n = 50/pool) for cDNA synthesis. Two different statistical algorithms geNorm and NormFinder were used for evaluating the stability of these genes. Based on gene stability measure (M value) and pairwise variation (V value), calculated by geNorm analysis, the most stable ICGs were RPS15, HPRT1, and ACTB for cloned blastocysts, HMBS, UBC, and HPRT1 for IVF blastocysts and RPS15, GAPDH, and HPRT1 for both the embryo types analyzed together. RPS18 was the least stable gene for both cloned and IVF blastocysts. Following NormFinder analysis, the order of stability was RPS15 = HPRT1>GAPDH for cloned blastocysts, HMBS = UBC>RPS23 for IVF blastocysts, and HPRT1>GAPDH>RPS15 for cloned and IVF blastocysts together. These results suggest that despite overlapping of the three most stable ICGs between cloned and IVF blastocysts, the panel of ICGs selected for normalization of qPCR data of cloned and IVF blastocyst-stage embryos should be different.

  19. Identification of human intestinal parasites affecting an asymptomatic peri-urban Argentinian population using multi-parallel quantitative real-time polymerase chain reaction.

    Science.gov (United States)

    Cimino, Rubén O; Jeun, Rebecca; Juarez, Marisa; Cajal, Pamela S; Vargas, Paola; Echazú, Adriana; Bryan, Patricia E; Nasser, Julio; Krolewiecki, Alejandro; Mejia, Rojelio

    2015-07-17

    In resource-limited countries, stool microscopy is the diagnostic test of choice for intestinal parasites (soil-transmitted helminths and/or intestinal protozoa). However, sensitivity and specificity is low. Improved diagnosis of intestinal parasites is especially important for accurate measurements of prevalence and intensity of infections in endemic areas. The study was carried out in Orán, Argentina. A total of 99 stool samples from a local surveillance campaign were analyzed by concentration microscopy and McMaster egg counting technique compared to the analysis by multi-parallel quantitative real-time polymerase chain reaction (qPCR). This study compared the performance of qPCR assay and stool microscopy for 8 common intestinal parasites that infect humans including the helminths Ascaris lumbricoides, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, and the protozoa Giardia lamblia, Cryptosporidium parvum/hominis, and Entamoeba histolytica, and investigated the prevalence of polyparasitism in an endemic area. qPCR showed higher detection rates for all parasites as compared to stool microscopy except T. trichiura. Species-specific primers and probes were able to distinguish between A. duodenale (19.1%) and N. americanus (36.4%) infections. There were 48.6% of subjects co-infected with both hookworms, and a significant increase in hookworm DNA for A. duodenale versus N. americanus (119.6 fg/μL: 0.63 fg/μL, P parasites in an endemic area that has improved diagnostic accuracy compared to stool microscopy. This first time use of multi-parallel qPCR in Argentina has demonstrated the high prevalence of intestinal parasites in a peri-urban area. These results will contribute to more accurate epidemiological survey, refined treatment strategies on a public scale, and better health outcomes in endemic settings.

  20. Adhesion of mutans streptococci to self-ligating ceramic brackets: in vivo quantitative analysis with real-time polymerase chain reaction.

    Science.gov (United States)

    Jung, Woo-Sun; Yang, Il-Hyung; Lim, Won Hee; Baek, Seung-Hak; Kim, Tae-Woo; Ahn, Sug-Joon

    2015-12-01

    To analyze in vivo mutans streptococci (MS) adhesion to self-ligating ceramic brackets [Clarity-SL (CSL) and Clippy-C (CC)] and the relationships between bacterial adhesion and oral hygiene indices. Four central incisor brackets from the maxilla and mandible were collected from 40 patients (20 patients per each bracket type) at debonding immediately after plaque and gingival indices were measured. Adhesions of Streptococcus mutans, S. sobrinus, and total bacteria were quantitatively determined using real-time polymerase chain reaction after genomic DNA was extracted. Factorial analysis of variance was used to analyze bacterial adhesion to the brackets with respect to the bracket type and jaw position. Correlation coefficients were calculated to determine the relationships of bacterial adhesion to oral hygiene indices. Adhesion of total bacteria and S. mutans to CSL was higher than that to CC (P brackets was higher than that to the maxillary ones (P brackets were higher than that in the mandibular ones (P brackets and jaw positions. Interestingly, no significant relationships were found between bacterial adhesions and oral hygiene indices. Complex bracket configurations may significantly influence bacterial adhesion to orthodontic brackets. Further in vivo study using bracket raw materials will help to define the relationships between bacteria adhesion and enamel demineralization. Because oral hygiene indices were not significantly correlated with adhesions of MS to self-ligating ceramic brackets, careful examinations around the brackets should be needed to prevent enamel demineralization, regardless of oral hygiene status. © The Author 2015. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  1. Identification of internal control genes for quantitative expression analysis by real-time PCR in bovine peripheral lymphocytes.

    Science.gov (United States)

    Spalenza, Veronica; Girolami, Flavia; Bevilacqua, Claudia; Riondato, Fulvio; Rasero, Roberto; Nebbia, Carlo; Sacchi, Paola; Martin, Patrice

    2011-09-01

    Gene expression studies in blood cells, particularly lymphocytes, are useful for monitoring potential exposure to toxicants or environmental pollutants in humans and livestock species. Quantitative PCR is the method of choice for obtaining accurate quantification of mRNA transcripts although variations in the amount of starting material, enzymatic efficiency, and the presence of inhibitors can lead to evaluation errors. As a result, normalization of data is of crucial importance. The most common approach is the use of endogenous reference genes as an internal control, whose expression should ideally not vary among individuals and under different experimental conditions. The accurate selection of reference genes is therefore an important step in interpreting quantitative PCR studies. Since no systematic investigation in bovine lymphocytes has been performed, the aim of the present study was to assess the expression stability of seven candidate reference genes in circulating lymphocytes collected from 15 dairy cows. Following the characterization by flow cytometric analysis of the cell populations obtained from blood through a density gradient procedure, three popular softwares were used to evaluate the gene expression data. The results showed that two genes are sufficient for normalization of quantitative PCR studies in cattle lymphocytes and that YWAHZ, S24 and PPIA are the most stable genes. Copyright © 2010 Elsevier Ltd. All rights reserved.

  2. Real-Time and High-Resolution 3D Face Measurement via a Smart Active Optical Sensor.

    Science.gov (United States)

    You, Yong; Shen, Yang; Zhang, Guocai; Xing, Xiuwen

    2017-03-31

    The 3D measuring range and accuracy in traditional active optical sensing, such as Fourier transform profilometry, are influenced by the zero frequency of the captured patterns. The phase-shifting technique is commonly applied to remove the zero component. However, this phase-shifting method must capture several fringe patterns with phase difference, thereby influencing the real-time performance. This study introduces a smart active optical sensor, in which a composite pattern is utilized. The composite pattern efficiently combines several phase-shifting fringes and carrier frequencies. The method can remove zero frequency by using only one pattern. Model face reconstruction and human face measurement were employed to study the validity and feasibility of this method. Results show no distinct decrease in the precision of the novel method unlike the traditional phase-shifting method. The texture mapping technique was utilized to reconstruct a nature-appearance 3D digital face.

  3. 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.

  4. A real time 155 GHz millimeter wave interferometer module for electron density measurement in large plasma devices

    International Nuclear Information System (INIS)

    Huettemann, P.W.; Waidmann, G.

    1982-09-01

    A homodyne, real time 155 GHz interferometer channel is described which is one module of a multichannel system for use on TEXTOR tokamak. A standing sine wave is generated in a phase bridge by transmitting a frequency modulated millimeter wave down two unequal interferometer branches. The presence of plasma produces a phase slip of the sine wave with respect to a reference signal. The phase shift is linear proportional to plasma density for expected TEXTOR plasmas. Long plasma paths give multiradian phase shifts which are recorded by a digital fringe counting system. The accuracy of phase measurement is ΔPHI = 2π/16. Phase changes of 7π/8 are accepted per modulation period. The microwave in the measurement branch of the interferometer is transmitted using a quasioptical technique. Components and technical details are described. The interferometer was tested in a simulation set-up and in two different plasma experiments. Experimental results are presented. (orig.)

  5. [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.

  6. Detection and semi-quantification of Strongylus vulgaris DNA in equine faeces by real-time quantitative PCR.

    Science.gov (United States)

    Nielsen, Martin K; Peterson, David S; Monrad, Jesper; Thamsborg, Stig M; Olsen, Susanne N; Kaplan, Ray M

    2008-03-01

    Strongylus vulgaris is an important strongyle nematode with high pathogenic potential infecting horses world-wide. Several decades of intensive anthelmintic use has virtually eliminated clinical disease caused by S. vulgaris, but has also caused high levels of anthelmintic resistance in equine small strongyle (cyathostomin) nematodes. Recommendations aimed at limiting the development of anthelmintic resistance by reducing treatment intensity raises a simultaneous demand for reliable and accurate diagnostic tools for detecting important parasitic pathogens. Presently, the only means available to differentiate among strongyle species in a faecal sample is by identifying individual L3 larvae following a two week coproculture procedure. The aim of the present study is to overcome this diagnostic obstacle by developing a fluorescence-based quantitative PCR assay capable of identifying S. vulgaris eggs in faecal samples from horses. Species-specific primers and a TaqMan probe were designed by alignment of published ribosomal DNA sequences of the second internal transcribed spacer of cyathostomin and Strongylus spp. nematodes. The assay was tested for specificity and optimized using genomic DNA extracted from identified male worms of Strongylus and cyathostomin species. In addition, eggs were collected from adult female worms and used to evaluate the quantitative potential of the assay. Statistically significant linear relationships were found between egg numbers and cycle of threshold (Ct) values. PCR results were unaffected by the presence of cyathostomin DNA in the sample and there was no indication of PCR inhibition by faecal sources. A field evaluation on faecal samples obtained from four Danish horse farms revealed a good agreement with the traditional larval culture (kappa-value=0.78), but with a significantly higher performance of the PCR assay. An association between Ct values and S. vulgaris larval counts was statistically significant. The present assay can

  7. Real-time PCR in detection and quantitation of Leishmania donovani for the diagnosis of Visceral Leishmaniasis patients and the monitoring of their response to treatment

    Science.gov (United States)

    Ghosh, Prakash; Khan, Md. Anik Ashfaq; Duthie, Malcolm S.; Vallur, Aarthy C.; Picone, Alessandro; Howard, Randall F.; Reed, Steven G.

    2017-01-01

    Sustained elimination of Visceral Leishmaniasis (VL) requires the reduction and control of parasite reservoirs to minimize the transmission of Leishmania donovani infection. A simple, reproducible and definitive diagnostic procedure is therefore indispensable for the early and accurate detection of parasites in VL, Relapsed VL (RVL) and Post Kala-azar Dermal Leishmaniasis (PKDL) patients, all of whom are potential reservoirs of Leishmania parasites. To overcome the limitations of current diagnostic approaches, a novel quantitative real-time polymerase chain reaction (qPCR) method based on Taqman chemistry was devised for the detection and quantification of L. donovani in blood and skin. The diagnostic efficacy was evaluated using archived peripheral blood buffy coat DNA from 40 VL, 40 PKDL, 10 RVL, 20 cured VL, and 40 cured PKDL along with 10 tuberculosis (TB) cases and 80 healthy endemic controls. Results were compared to those obtained using a Leishmania-specific nested PCR (Ln-PCR). The real time PCR assay was 100% (95% CI, 91.19–100%) sensitive in detecting parasite genomes in VL and RVL samples and 85.0% (95% CI, 70.16–94.29%) sensitive for PKDL samples. In contrast, the sensitivity of Ln-PCR was 77.5% (95% CI, 61.55–89.16%) for VL samples, 100% (95%CI, 69.15–100%) for RVL samples, and 52.5% (95% CI, 36.13–68.49%) for PKDL samples. There was significant discordance between the two methods with the overall sensitivity of the qPCR assay being considerably higher than Ln-PCR. None of the assay detected L. donovani DNA in buffy coats from cured VL cases, and reduced infectious burdens were demonstrated in cured PKDL cases who remained positive in 7.5% (3/40) and 2.5% (1/40) cases by real-time PCR and Ln-PCR, respectively. Both assays were 100% (95% CI, 95.98–100) specific with no positive signals in either endemic healthy control or TB samples. The real time PCR assay we developed offers a molecular tool for accurate detection of circulating L

  8. 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.

  9. 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.

  10. Selection of internal control genes for real-time quantitative PCR in ovary and uterus of sows across pregnancy.

    Directory of Open Access Journals (Sweden)

    María Martínez-Giner

    Full Text Available BACKGROUND: Reproductive traits play a key role in pig production in order to reduce costs and increase economic returns. Among others, gene expression analyses represent a useful approach to study genetic mechanisms underlying reproductive traits in pigs. The application of reverse-transcription quantitative PCR requires the selection of appropriate reference genes, whose expression levels should not be affected by the experimental conditions, especially when comparing gene expression across different physiological stages. RESULTS: The gene expression stability of ten potential reference genes was studied by three different methods (geNorm, NormFinder and BestKeeper in ovary and uterus collected at five different physiological time points (heat, and 15, 30, 45 and 60 days of pregnancy. Although final ranking differed, the three algorithms gave very similar results. Thus, the most stable genes across time were TBP and UBC in uterus and TBP and HPRT1 in ovary, while HMBS and ACTB showed the less stable expression in uterus and ovary, respectively. When studied as a systematic effect, the reproductive stage did not significantly affect the expression of the candidate reference genes except at 30d and 60d of pregnancy, when a general drop in expression was observed in ovary. CONCLUSIONS: Based in our results, we propose the use of TBP, UBC and SDHA in uterus and TBP, GNB2L1 and HPRT1 in ovary for normalization of longitudinal expression studies using quantitative PCR in sows.

  11. 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....

  12. 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...

  13. Combining real-time PCR and next-generation DNA sequencing to provide quantitative comparisons of fungal aerosol populations

    Science.gov (United States)

    Dannemiller, Karen C.; Lang-Yona, Naama; Yamamoto, Naomichi; Rudich, Yinon; Peccia, Jordan

    2014-02-01

    We examined fungal communities associated with the PM10 mass of Rehovot, Israel outdoor air samples collected in the spring and fall seasons. Fungal communities were described by 454 pyrosequencing of the internal transcribed spacer (ITS) region of the fungal ribosomal RNA encoding gene. To allow for a more quantitative comparison of fungal exposure in humans, the relative abundance values of specific taxa were transformed to absolute concentrations through multiplying these values by the sample's total fungal spore concentration (derived from universal fungal qPCR). Next, the sequencing-based absolute concentrations for Alternaria alternata, Cladosporium cladosporioides, Epicoccum nigrum, and Penicillium/Aspergillus spp. were compared to taxon-specific qPCR concentrations for A. alternata, C. cladosporioides, E. nigrum, and Penicillium/Aspergillus spp. derived from the same spring and fall aerosol samples. Results of these comparisons showed that the absolute concentration values generated from pyrosequencing were strongly associated with the concentration values derived from taxon-specific qPCR (for all four species, p 0.70). The correlation coefficients were greater for species present in higher concentrations. Our microbial aerosol population analyses demonstrated that fungal diversity (number of fungal operational taxonomic units) was higher in the spring compared to the fall (p = 0.02), and principal coordinate analysis showed distinct seasonal differences in taxa distribution (ANOSIM p = 0.004). Among genera containing allergenic and/or pathogenic species, the absolute concentrations of Alternaria, Aspergillus, Fusarium, and Cladosporium were greater in the fall, while Cryptococcus, Penicillium, and Ulocladium concentrations were greater in the spring. The transformation of pyrosequencing fungal population relative abundance data to absolute concentrations can improve next-generation DNA sequencing-based quantitative aerosol exposure assessment.

  14. Total neutron-counting plutonium inventory measurement systems (PIMS) and their potential application to near real time materials accountancy (NRTMA)

    International Nuclear Information System (INIS)

    Driscall, I.; Fox, G.H.; Orr, C.H.; Whitehouse, K.R.

    1988-01-01

    A radiometric method of determining the inventory of an operating plutonium plant is described. An array of total neutron counters distributed across the plant is used to estimate hold-up at each plant item. Corrections for the sensitivity of detectors to plutonium in adjacent plant items are achieved through a matrix approach. This paper describes our experience in design, calibration and operation of a Plutonium Inventory Measurement System (PIMS) on an oxalate precipitation plutonium finishing line. Data from a recent trial of Near-Real-Time Materials Accounting (NRTMA) using the PIMS are presented and used to illustrate its present performance and problem areas. The reader is asked to consider what role PIMS might have in future accountancy systems

  15. 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®.

  16. Identification of appropriate reference genes for human mesenchymal stem cell analysis by quantitative real-time PCR.

    Science.gov (United States)

    Li, Xiuying; Yang, Qiwei; Bai, Jinping; Xuan, Yali; Wang, Yimin

    2015-01-01

    Normalization to a reference gene is the method of choice for quantitative reverse transcription-PCR (RT-qPCR) analysis. The stability of reference genes is critical for accurate experimental results and conclusions. We have evaluated the expression stability of eight commonly used reference genes found in four different human mesenchymal stem cells (MSC). Using geNorm, NormFinder and BestKeeper algorithms, we show that beta-2-microglobulin and peptidyl-prolylisomerase A were the optimal reference genes for normalizing RT-qPCR data obtained from MSC, whereas the TATA box binding protein was not suitable due to its extensive variability in expression. Our findings emphasize the significance of validating reference genes for qPCR analyses. We offer a short list of reference genes to use for normalization and recommend some commercially-available software programs as a rapid approach to validate reference genes. We also demonstrate that the two reference genes, β-actin and glyceraldehyde-3-phosphate dehydrogenase, are frequently used are not always successful in many cases.

  17. Selection of reference genes for quantitative real time RT-PCR during dimorphism in the zygomycete Mucor circinelloides.

    Science.gov (United States)

    Valle-Maldonado, Marco I; Jácome-Galarza, Irvin E; Gutiérrez-Corona, Félix; Ramírez-Díaz, Martha I; Campos-García, Jesús; Meza-Carmen, Víctor

    2015-03-01

    Mucor circinelloides is a dimorphic fungal model for studying several biological processes including cell differentiation (yeast-mold transitions) as well as biodiesel and carotene production. The recent release of the first draft sequence of the M. circinelloides genome, combined with the availability of analytical methods to determine patterns of gene expression, such as quantitative Reverse transcription-Polymerase chain reaction (qRT-PCR), and the development of molecular genetic tools for the manipulation of the fungus, may help identify M. circinelloides gene products and analyze their relevance in different biological processes. However, no information is available on M. circinelloides genes of stable expression that could serve as internal references in qRT-PCR analyses. One approach to solve this problem consists in the use of housekeeping genes as internal references. However, validation of the usability of these reference genes is a fundamental step prior to initiating qRT-PCR assays. This work evaluates expression of several constitutive genes by qRT-PCR throughout the morphological differentiation stages of M. circinelloides; our results indicate that tfc-1 and ef-1 are the most stable genes for qRT-PCR assays during differentiation studies and they are proposed as reference genes to carry out gene expression studies in this fungus.

  18. Real time Faraday spectrometer

    Science.gov (United States)

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

    1991-01-01

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

  19. Real time Faraday spectrometer

    International Nuclear Information System (INIS)

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

    1991-01-01

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

  20. Use of REMPI-TOFMS for real-time measurement of trace aromatics during operation of aircraft ground equipment

    Science.gov (United States)

    Gullett, Brian; Touati, Abderrahmane; Oudejans, Lukas

    Emissions of aromatic air toxics from aircraft ground equipment (AGE) were measured with a resonance enhanced multiphoton ionization-time of flight mass spectrometry (REMPI-TOFMS) system consisting of a pulsed solid state laser for photoionization and a TOFMS for mass discrimination. This instrument was capable of characterizing turbine emissions and the effect of varying load operations on pollutant production. REMPI-TOFMS is capable of high selectivity and low detection limits (part per trillion to part per billion) in real time (1 s resolution). Hazardous air pollutants and criteria pollutants were measured during startups and idle and full load operations. Measurements of compounds such as benzene, toluene, ethylbenzene, xylenes, styrene, and polycyclic aromatic hydrocarbons compared well with standard methods. Startup emissions from the AGE data showed persistent concentrations of pollutants, unlike those from a diesel generator, where a sharp spike in emissions rapidly declined to steady state levels. The time-resolved responses of air toxics concentrations varied significantly by source, complicating efforts to minimize these emissions with common operating prescriptions. The time-resolved measurements showed that pollutant concentrations decline (up to 5×) in a species-specific manner over the course of multiple hours of operation, complicating determination of accurate and precise emission factors via standard extractive sampling. Correlations of air toxic concentrations with more commonly measured pollutants such as CO or PM were poor due to the relatively greater changes in the measured toxics' concentrations.

  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.

  2. 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

  3. 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

  4. The Smallest R/V: A Small-scale Ocean Exploration Demonstration of Real-time Bathymetric Measurements

    Science.gov (United States)

    Howell, S. M.; Boston, B.; Maher, S. M.; Sleeper, J. D.; Togia, H.; Tree, J. P.

    2014-12-01

    In October 2013, graduate student members of the University of Hawaii Geophysical Society designed a small-scale model research vessel (R/V) that uses sonar to create 3D maps of a model seafloor in real-time. This pilot project was presented to the public at the School of Ocean and Earth Science and Technology's (SOEST) Biennial Open House weekend. An estimated 7,600 people attended the two-day event, including children and teachers from Hawaii's schools, home school students, community groups, families, and science enthusiasts. Our exhibit demonstrated real-time sonar mapping of a cardboard volcano using a toy size research vessel on a fixed 2D model ship track suspended above a model seafloor. Sound wave travel times were recorded using an unltrasonic emitter/receiver attached to an Arduino microcontroller platform, while the same system measured displacement along the ship track. This data was streamed through a USB connection to a PC running MatLab, where a 3D model was updated as the ship collected data. Our exhibit demonstrates the practical use of complicated concepts, like wave physics and data processing, in a way that even the youngest elementary students are able to understand. It provides an accessible avenue to learn about sonar mapping, and could easily be adapted to talk about bat and marine mammal echolocation by replacing the model ship and volcano. The exhibit received an overwhelmingly positive response from attendees, and has inspired the group to develop a more interactive model for future exhibitions, using multiple objects to be mapped that participants could arrange, and a more robust ship movement system that participants could operate.

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

    Science.gov (United States)

    Droplet digital Polymerase chain reaction (ddPCR) is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. It is a promising DNA quantification technology for medical diagnostics but there are only a few reports of its use for plant pat...

  6. A digital, constant-frequency pulsed phase-locked-loop instrument for real-time, absolute ultrasonic phase measurements

    Science.gov (United States)

    Haldren, H. A.; Perey, D. F.; Yost, W. T.; Cramer, K. E.; Gupta, M. C.

    2018-05-01

    A digitally controlled instrument for conducting single-frequency and swept-frequency ultrasonic phase measurements has been developed based on a constant-frequency pulsed phase-locked-loop (CFPPLL) design. This instrument uses a pair of direct digital synthesizers to generate an ultrasonically transceived tone-burst and an internal reference wave for phase comparison. Real-time, constant-frequency phase tracking in an interrogated specimen is possible with a resolution of 0.000 38 rad (0.022°), and swept-frequency phase measurements can be obtained. Using phase measurements, an absolute thickness in borosilicate glass is presented to show the instrument's efficacy, and these results are compared to conventional ultrasonic pulse-echo time-of-flight (ToF) measurements. The newly developed instrument predicted the thickness with a mean error of -0.04 μm and a standard deviation of error of 1.35 μm. Additionally, the CFPPLL instrument shows a lower measured phase error in the absence of changing temperature and couplant thickness than high-resolution cross-correlation ToF measurements at a similar signal-to-noise ratio. By showing higher accuracy and precision than conventional pulse-echo ToF measurements and lower phase errors than cross-correlation ToF measurements, the new digitally controlled CFPPLL instrument provides high-resolution absolute ultrasonic velocity or path-length measurements in solids or liquids, as well as tracking of material property changes with high sensitivity. The ability to obtain absolute phase measurements allows for many new applications than possible with previous ultrasonic pulsed phase-locked loop instruments. In addition to improved resolution, swept-frequency phase measurements add useful capability in measuring properties of layered structures, such as bonded joints, or materials which exhibit non-linear frequency-dependent behavior, such as dispersive media.

  7. Characterization of PEBBLEs as a Tool for Real-Time Measurement of Dictyostelium discoideum Endosomal pH

    Directory of Open Access Journals (Sweden)

    Everett Moding

    2009-01-01

    Full Text Available The measurement of intracellular ion concentration change is important for understanding the cellular mechanisms for communication. Recently developed nanosensors, (Photonic Explorers for Biomedical use with Biologically Localized Embedding PEBBLEs, have a number of advantages for measuring ions in cells over established methods using microelectrodes, unbound fluorescent dyes, or NMR. PEBBLE sensors have been shown to work in principle for measuring dynamic ion changes, but few in vivo applications have been demonstrated. We modified the protocol for the fabrication of pH sensing PEBBLEs and developed a protocol for the utilization of these sensors for the monitoring of dynamic pH changes in the endosomes of slime mold Dictyostelium discoideum (D. discoideum. Oregon Green 514-CdSe Quantum Dot PEBBLEs were used to measure real-time pH inside D. discoideum endosomes during cAMP stimulation. Endosomal pH was shown to decrease during cAMP signaling, demonstrating a movement of protons into the endosomes of D. discoideum amoebae.

  8. Real-time measurement of plutonium in air by direct-inlet surface ionization mass spectrometry. Status report

    International Nuclear Information System (INIS)

    Stoffels, J.J.

    1980-04-01

    A new technique is being developed for monitoring low-level airborne plutonium on a real-time basis. The technique is based on surface ionization mass spectrometry of airborne particles. It will be capable of measuring plutonium concentrations below the maximum permissible concentration (MPC) level. A complete mass spectrometer was designed and constructed for this purpose. Major components which were developed and made operational for the instrument include an efficient inlet for directly sampling particles in air, a wide dynamic range ion detector and a minicomputer-based ion-burst measurement system. Calibration of the direct-inlet mass spectrometer (DIMS) was initiated to establish the instrument's response to plutonium dioxide as a function of concentration and particle size. This work revealed an important problem - bouncing of particles upon impact with the ionizing filament. Particle bounce results in a significant loss of measurement sensitivity. The feasibility of using an oven ionizer to overcome the particle bounce problem has been demonstrated. A rhenium oven ionizer was designed and constructed for the purpose of trapping particles which enter via the direct inlet. High-speed particles were trapped in the oven yielding a measurement sensitivity comparable to that for particles which are preloaded. Development of the Pu DIMS can now be completed by optimizing the oven design and calibrating the instrument's performance with UO 2 and CeO 2 particles as analogs to PuO 2 particles

  9. 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.

  10. Quantitative analysis of the improvement in omnidirectional maritime surveillance and tracking due to real-time image enhancement

    Science.gov (United States)

    de Villiers, Jason P.; Bachoo, Asheer K.; Nicolls, Fred C.; le Roux, Francois P. J.

    2011-05-01

    Tracking targets in a panoramic image is in many senses the inverse problem of tracking targets with a narrow field of view camera on a pan-tilt pedestal. In a narrow field of view camera tracking a moving target, the object is constant and the background is changing. A panoramic camera is able to model the entire scene, or background, and those areas it cannot model well are the potential targets and typically subtended far fewer pixels in the panoramic view compared to the narrow field of view. The outputs of an outward staring array of calibrated machine vision cameras are stitched into a single omnidirectional panorama and used to observe False Bay near Simon's Town, South Africa. A ground truth data-set was created by geo-aligning the camera array and placing a differential global position system receiver on a small target boat thus allowing its position in the array's field of view to be determined. Common tracking techniques including level-sets, Kalman filters and particle filters were implemented to run on the central processing unit of the tracking computer. Image enhancement techniques including multi-scale tone mapping, interpolated local histogram equalisation and several sharpening techniques were implemented on the graphics processing unit. An objective measurement of each tracking algorithm's robustness in the presence of sea-glint, low contrast visibility and sea clutter - such as white caps is performed on the raw recorded video data. These results are then compared to those obtained with the enhanced video data.

  11. Multi-site magnetotelluric measurement system with real-time data analysis. Final technical report No. 210

    Energy Technology Data Exchange (ETDEWEB)

    Becker, J.D.; Bostick, F.X. Jr.; Smith, H.W.

    1981-09-01

    A magnetotelluric measurement system has been designed to provide a more cost effective electrical method for geothermal and mineral exploration. The theoretical requirements and sensitivities of the magnetotelluric inversion process were specifically addressed in determining system performance requirements. Significantly reduced instrument noise levels provide improved data quality, and